From twheelock <@t> mclean.harvard.edu Thu Sep 1 09:42:39 2005 From: twheelock <@t> mclean.harvard.edu (Tim Wheelock) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: <3F3035CC.5090101@mclean.harvard.edu> Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 From Terry.Marshall <@t> rothgen.nhs.uk Thu Sep 1 09:49:01 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: Get the silly woman to wear sensible shoes:-) People who skate on thin ice cannot complain if they fall in. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Tim Wheelock [mailto:twheelock@mclean.harvard.edu] Sent: 05 August 2003 23:55 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] SLIPPING ON LAB FLOOR Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Thu Sep 1 10:03:15 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: We put rubber backed area rugs on the main problem areas. There's still some slipperiness, but it's much better than before. Another solution is replacing the tiles with non-slip tiles. They're kind of ugly but work very well. Third option, keep some sneakers in a drawer for her. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tim Wheelock Sent: Tuesday, August 05, 2003 5:55 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] SLIPPING ON LAB FLOOR Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From celebrej <@t> HHSC.CA Thu Sep 1 10:08:04 2005 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: <3AADFB88753AD31189C100902786B91C14E6E1EE@hch_nt_exchange.hhsc.ca> Hi Tim, We had similar problems over the years, and what we've done is placed mats on our floors along with a sign to warn all. Housekeeping ends up vacuuming the mats nightly so they don't end up looking messy. Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca -----Original Message----- From: Tim Wheelock [mailto:twheelock@mclean.harvard.edu] Sent: Tuesday, August 05, 2003 6:55 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] SLIPPING ON LAB FLOOR Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The Cornerstone of Care Campaign for Hamilton Health Sciences needs your support. Please, visit hamiltonhealthsciences.ca today and help make something great even greater. This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From cormier <@t> MIT.EDU Thu Sep 1 10:30:51 2005 From: cormier <@t> MIT.EDU (Kathy Cormier) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR In-Reply-To: Message-ID: <5.2.1.1.2.20050901112131.0112c068@po14.mit.edu> Hi Tim, All the responses that I have seen so far are things that I have seen done in the Histology labs that I have worked in, and they all seem to improve the situation. What we do here (and is WAY more fun) is to order "sticky mats", just like they use in animal clean rooms or manufacturing of computer or other sensitive machinery. We get ours through VWR the frame is # 21924-832 (about $55.00 and reusable) and the multi layer mats are # 21924-840 ($136.00, 60 peel away layers, I think 4 mats per case) We use roughly a case a year and the layers get peeled about once a week. It's like human flypaper, really fun on the unsuspecting.... Kathy Cormier Histology Necropsy Supervisor Division of Comparative Medicine MIT >-----Original Message----- >From: Tim Wheelock [mailto:twheelock@mclean.harvard.edu] >Sent: 05 August 2003 23:55 >To: 'histonet@lists.utsouthwestern.edu' >Subject: [Histonet] SLIPPING ON LAB FLOOR > > >Hi Everyone: > >A problem. >My boss, who wears dress shoes with half-size high heels (the heels have >a broad base to them, not the "stiletto" type), has been having problems >slipping on my laboratory's floor. I am really afraid that she is going >to actually fall and injure herself. > >I myself have no problem in the lab, since the soles of my shoes are >rubber or plastic polymer. Sneakers work fine as well. >I manage a neuropathology laboratory which means I use paraffin embedding. >Although, I keep the floor clean, I think that the residual wax near the >embedding and sectioning stations may get spread around the rest of the >lab by my shoes. > >So far, I have put a "CAUTION" sign up on my laboratory door advising >people to excercise caution when entering the lab, especially when >wearing dress shoes, in order to at least increase awareness. > >Perhaps, I should put the laboratory floor on a regular "preventative >maintenance" schedule of cleaning and waxing to minimize the amount of >wax on the floor. >Then again, maybe I should ask the maintenance people not to put any wax >on the floor after they clean it. Perhaps it is this wax that is part of >the problem. > >Has anyone ever had this problem? How did you solve it? > >I would appreciate any advice anyone may have. > >Tim Wheelock >Harvard Brain Tissue Resource Center >McLean Hospital >617-855-3592 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Thu Sep 1 10:34:19 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB43B8@fh2k093.fhmis.net> Get some rubber mats SOON!! You can find them with Safety Supply Corp. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: 'histonet@lists.utsouthwestern.edu' Sent: 8/5/2003 6:55 PM Subject: [Histonet] SLIPPING ON LAB FLOOR Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Thu Sep 1 10:43:03 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Animal Information for the Hurricane Area Message-ID: <6.1.1.1.2.20050901113554.019bd210@mail.vet.upenn.edu> Good Morning To All, I am forwarding the text of a message we received this morning at the Vet School. We are all being made aware of where to send money (preferably) to assist the human misery in the area Katrina had destroyed, especially in New Orleans. However, many animals were left behind and are in trouble also. This was the first message I had seen informing those who wish to help the animals or know what is being done for them. I hope this does not offend anyone and the I realize the human suffering is so great that this can seem trivial to some. >Dear Faculty & Staff: > >As you know, Hurricane Katrina, which struck the Gulf Coast and the city of >New Orleans, LA, this week, has tragically claimed lives and left thousands >more homeless. As rescue workers continue to aid those in need, many of you >have expressed concern regarding the various animals that were stranded as >a result of this disaster. Please be assured that there are several >organizations working to rescue and treat as many animals as possible. > >The AVMA Veterinary Medical Assistance Team and the Houston SPCA are >working together to set up a displaced animal shelter on the Baton Rouge >campus of Louisiana State University and at another site a few miles from >Baton Rouge. If you would like to contribute to this effort, please send a >check to The Walter J. Ernst Veterinary Memorial Foundation and write >Disaster Relief Fund on the memo line. Donations should be sent to the >LVMA, 8550 United Plaza Blvd, Suite 1001, Baton Rouge LA 70809. In >addition, the PVMA Foundation is likely to set up a fund for this effort. > >Other organizations accepting donations for animal rescue and welfare are: > * Noah's Wish (www.noahswish.org) > * ASPCA (www.aspca.org) > * AVMA (www.avma.org) > * Best Friends Animal Society >(www.bestfriends.org) > * The Humane Society of the United States >(www.hsus.org) > >Thank you for your support of these important efforts. > > Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From gcallis <@t> montana.edu Thu Sep 1 11:06:53 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Re: SLIPPING ON LAB FLOOR In-Reply-To: <3F3035CC.5090101@mclean.harvard.edu> References: <3F3035CC.5090101@mclean.harvard.edu> Message-ID: <6.0.0.22.1.20050901095114.01b0cce8@gemini.msu.montana.edu> She is putting herself at risk for the sake of being a glamourous professional. Been there, did this in 60's - but with stilettos and very pointy toe styles, and suffered through the "slip sliding away" thing in a histology laboratory. It took a near disaster to teach an eyeopening safety awareness lesson and buying sensible safe shoes. One of the most common accidents in the work place is falling. She should use practical ( and some are not that ugly!) work related shoes, slip on style, for daily wear in office/lab, and change in a flash when she needs to be the "dressed to the nines" professional for meetings, conferences, etc. You can minimize paraffin debris by putting sticky mats (3M, sold by vendors) under your paraffin embedding, microtomy and processing areas, and in front of doorways. We put industrial rug/mats in those areas but clinical labs may have a hard time with cleaning. Our chairs roll over the mats easily, and they can be vacuumed without difficulty. At 04:55 PM 8/5/2003, you wrote: >Hi Everyone: > >A problem. >My boss, who wears dress shoes with half-size high heels (the heels have a >broad base to them, not the "stiletto" type), has been having problems >slipping on my laboratory's floor. I am really afraid that she is going to >actually fall and injure herself. > >I myself have no problem in the lab, since the soles of my shoes are >rubber or plastic polymer. Sneakers work fine as well. >I manage a neuropathology laboratory which means I use paraffin embedding. >Although, I keep the floor clean, I think that the residual wax near the >embedding and sectioning stations may get spread around the rest of the >lab by my shoes. > >So far, I have put a "CAUTION" sign up on my laboratory door advising >people to excercise caution when entering the lab, especially when wearing >dress shoes, in order to at least increase awareness. > >Perhaps, I should put the laboratory floor on a regular "preventative >maintenance" schedule of cleaning and waxing to minimize the amount of wax >on the floor. >Then again, maybe I should ask the maintenance people not to put any wax >on the floor after they clean it. Perhaps it is this wax that is part of >the problem. > >Has anyone ever had this problem? How did you solve it? > >I would appreciate any advice anyone may have. > >Tim Wheelock >Harvard Brain Tissue Resource Center >McLean Hospital >617-855-3592 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From lrichey <@t> u.washington.edu Thu Sep 1 11:14:45 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] (no subject) In-Reply-To: <1125504489.4315d5e9617de@webmail.shef.ac.uk> References: <1125504489.4315d5e9617de@webmail.shef.ac.uk> Message-ID: <431728F5.9070300@u.washington.edu> We fix all precut control tissue slides in cold acetone for 5 minutes,and allow them to air dry before storing them in the -70. Matt Prideaux wrote: >Dear Histonetters, > >A colleague wishes to store cytospun cell preparations in the freezer before >doing immunocytochemistry for CD31 and CD44. She would like to know whether >it's better to fix the slides before freezing or after thawing them prior to >staining. Also, which fixatives would you recommend? > >Matt > > From jqb7 <@t> cdc.gov Thu Sep 1 11:04:41 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: It may not necessarily the dress shoes. None of the shoes I wear are such but one pair tend to slip on the paraffin-coated floors more than the others. Also, we often have non-lab personnel enter the lab for a variety of reasons, for example our secretary. The new non-slip mats would probably be a big help. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, September 01, 2005 10:49 AM To: Tim Wheelock; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR Get the silly woman to wear sensible shoes:-) People who skate on thin ice cannot complain if they fall in. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Tim Wheelock [mailto:twheelock@mclean.harvard.edu] Sent: 05 August 2003 23:55 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] SLIPPING ON LAB FLOOR Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kweidenh <@t> montefiore.org Thu Sep 1 11:22:00 2005 From: kweidenh <@t> montefiore.org (Karen Weidenheim) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: Dear HIstonet, This is certainly a problem. As a neuropathologist I almost always wear flat rubber/polymer soled shoes at work because of the problem Tim has noted. Flat rubber/polymer soled shoes that look reasonably professional are available. If I think I just must wear dress shoes for some meeting or conference I bring them to work and put them on for that purpose only. You might suggest this to your boss. One is also supposed to wear closed toe shoes in the lab at least in New York State--another thing most attendings and a few techs ignore. The rule is meant to protect your feet if you spill formalin, bleach, acid etc. Having spilled on myself before, I also suggest this is a good idea. I have known at least one (very fashionable) pathologist who broke her ankle due to paraffin on the floor. After that they carpeted the corridor outside the lab and put in a schedule of regular stripping for the lab floor itself. I think safety is more important than looks in the path lab and I think any dress code that one's hospital may have should consider what we work around when they judge our dress. Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> Tim Wheelock 8/5/2003 6:55:08 PM >>> Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From STEGTM <@t> samcstl.org Thu Sep 1 11:30:40 2005 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] sentinel biopsies Message-ID: We place the specimen containers in a lead cabinet for a day, then process as usual. From 1dpeterson <@t> meriter.com Thu Sep 1 11:54:58 2005 From: 1dpeterson <@t> meriter.com (Peterson, Dan) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Hurricane Relief Message-ID: <328CBAE62F31C642B422970E879DFADC01A7FFCA@pcwex01> With everybody's thoughts and prayers being sent to Katrina's victims, wouldn't it be nice to send a little financial aid also? Just a thought, why not have a collection point at the upcoming NSH convention, maybe the Outpost? With the number of members that will be attending (and maybe collecting from their co-workers before hand) a fair amount could be collected and donated to (?) by the NSH on behalf of Histologists nationwide. Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories (608)-267-6557 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. From jnocito <@t> satx.rr.com Thu Sep 1 12:00:07 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR References: <3F3035CC.5090101@mclean.harvard.edu> Message-ID: <002e01c5af16$9b5cd1e0$52bf0b43@yourxhtr8hvc4p> right now, we have a pool going to see who falls first, one of the billing people or one of our sales reps (I'm betting on the sales rep) only kidding, only kidding. We have signs up and everyone knows to be careful when they come in the lab. The CEO has told them to be careful too. But, that is only a temporary problem. In our new lab, we are having the blocks and slides files outside of the lab. We also are having a new floor put in that has skid-proof material embedded in the concrete. Slipping shouldn't be a problem. That's if we ever move into the new building. Joe " The Toe" Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Tim Wheelock" To: Sent: Tuesday, August 05, 2003 5:55 PM Subject: [Histonet] SLIPPING ON LAB FLOOR > Hi Everyone: > > A problem. > My boss, who wears dress shoes with half-size high heels (the heels have a > broad base to them, not the "stiletto" type), has been having problems > slipping on my laboratory's floor. I am really afraid that she is going to > actually fall and injure herself. > > I myself have no problem in the lab, since the soles of my shoes are > rubber or plastic polymer. Sneakers work fine as well. > I manage a neuropathology laboratory which means I use paraffin embedding. > Although, I keep the floor clean, I think that the residual wax near the > embedding and sectioning stations may get spread around the rest of the > lab by my shoes. > > So far, I have put a "CAUTION" sign up on my laboratory door advising > people to excercise caution when entering the lab, especially when wearing > dress shoes, in order to at least increase awareness. > > Perhaps, I should put the laboratory floor on a regular "preventative > maintenance" schedule of cleaning and waxing to minimize the amount of wax > on the floor. > Then again, maybe I should ask the maintenance people not to put any wax > on the floor after they clean it. Perhaps it is this wax that is part of > the problem. > > Has anyone ever had this problem? How did you solve it? > > I would appreciate any advice anyone may have. > > Tim Wheelock > Harvard Brain Tissue Resource Center > McLean Hospital > 617-855-3592 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.344 / Virus Database: 267.10.18/86 - Release Date: 8/31/2005 > From dmarsha3 <@t> utmem.edu Thu Sep 1 12:01:27 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Animal Information for the Hurricane Area References: <6.1.1.1.2.20050901113554.019bd210@mail.vet.upenn.edu> Message-ID: <000601c5af16$cb50f930$f623c084@DanaM> thanks pamela, if everyone cares some, then there caring enough for all living things.......... people and critters dm ----- Original Message ----- From: "Pamela Marcum" To: Sent: Thursday, September 01, 2005 10:43 AM Subject: [Histonet] Animal Information for the Hurricane Area > Good Morning To All, > > I am forwarding the text of a message we received this morning at the Vet > School. We are all being made aware of where to send money (preferably) > to assist the human misery in the area Katrina had destroyed, especially > in New Orleans. However, many animals were left behind and are in trouble > also. This was the first message I had seen informing those who wish to > help the animals or know what is being done for them. I hope this does not > offend anyone and the I realize the human suffering is so great that this > can seem trivial to some. > > >>Dear Faculty & Staff: >> >>As you know, Hurricane Katrina, which struck the Gulf Coast and the city >>of >>New Orleans, LA, this week, has tragically claimed lives and left >>thousands >>more homeless. As rescue workers continue to aid those in need, many of >>you >>have expressed concern regarding the various animals that were stranded as >>a result of this disaster. Please be assured that there are several >>organizations working to rescue and treat as many animals as possible. >> >>The AVMA Veterinary Medical Assistance Team and the Houston SPCA are >>working together to set up a displaced animal shelter on the Baton Rouge >>campus of Louisiana State University and at another site a few miles from >>Baton Rouge. If you would like to contribute to this effort, please send a >>check to The Walter J. Ernst Veterinary Memorial Foundation and write >>Disaster Relief Fund on the memo line. Donations should be sent to the >>LVMA, 8550 United Plaza Blvd, Suite 1001, Baton Rouge LA 70809. In >>addition, the PVMA Foundation is likely to set up a fund for this effort. >> >>Other organizations accepting donations for animal rescue and welfare are: >> * Noah's Wish (www.noahswish.org) >> * ASPCA (www.aspca.org) >> * AVMA (www.avma.org) >> * Best Friends Animal Society >>(www.bestfriends.org) >> * The Humane Society of the United States >>(www.hsus.org) >> >>Thank you for your support of these important efforts. >> >> > > Best Regards, > > Pamela A Marcum > Manager, Histology Special Procedures > University of Pennsylvania > School of Veterinary Medicine > R.S. Reynolds Jr. CORL > New Bolton Center > 382 West Street Road > Kennett Square, PA 19348 > > Phone - 610-925-6278 > Fax - 610-925-8120 > E-mail - pmarcum@vet.upenn.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From NORMANB <@t> ccf.org Thu Sep 1 12:15:07 2005 From: NORMANB <@t> ccf.org (Norman, Barbara) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Hurricane Relief Message-ID: <9C6B798812BF1041B73D84670240C8060B3FCE@cchsncexmb51.nc.ad.cchs.net> Sounds great, NSH is working on that as we speak, I'm sure more details will follow in the next few days. Bless the American Spirit! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Peterson, Dan Sent: Thursday, September 01, 2005 12:55 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hurricane Relief With everybody's thoughts and prayers being sent to Katrina's victims, wouldn't it be nice to send a little financial aid also? Just a thought, why not have a collection point at the upcoming NSH convention, maybe the Outpost? With the number of members that will be attending (and maybe collecting from their co-workers before hand) a fair amount could be collected and donated to (?) by the NSH on behalf of Histologists nationwide. Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories (608)-267-6557 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sfarley <@t> seattlecca.org Thu Sep 1 12:53:48 2005 From: sfarley <@t> seattlecca.org (Farley, Sunni R) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Warthin-Starry Technique Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B94832340E@wala01.seattlecca.org> I was just wondering if anybody regularly uses the Warthin-Starry technique for spirochetes and wouldn't mind forwarding me a copy of their procedure. Thank you in advance for your help! Sunni R. Farley Histotechnician Clinical Pathology Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From Janet.Bonner <@t> FLHOSP.ORG Thu Sep 1 13:08:47 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Re: SLIPPING ON LAB FLOOR Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB43BD@fh2k093.fhmis.net> Actually, you should put some kind of warning to this effect in writing and deliver it to all in the laboratory as well as put a warning to that effect on the door. This will cover you incase of a resulting law suit. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Tim Wheelock; Histonet@lists.utsouthwestern.edu Sent: 9/1/2005 12:06 PM Subject: [Histonet] Re: SLIPPING ON LAB FLOOR She is putting herself at risk for the sake of being a glamourous professional. Been there, did this in 60's - but with stilettos and very pointy toe styles, and suffered through the "slip sliding away" thing in a histology laboratory. It took a near disaster to teach an eyeopening safety awareness lesson and buying sensible safe shoes. One of the most common accidents in the work place is falling. She should use practical ( and some are not that ugly!) work related shoes, slip on style, for daily wear in office/lab, and change in a flash when she needs to be the "dressed to the nines" professional for meetings, conferences, etc. You can minimize paraffin debris by putting sticky mats (3M, sold by vendors) under your paraffin embedding, microtomy and processing areas, and in front of doorways. We put industrial rug/mats in those areas but clinical labs may have a hard time with cleaning. Our chairs roll over the mats easily, and they can be vacuumed without difficulty. At 04:55 PM 8/5/2003, you wrote: >Hi Everyone: > >A problem. >My boss, who wears dress shoes with half-size high heels (the heels have a >broad base to them, not the "stiletto" type), has been having problems >slipping on my laboratory's floor. I am really afraid that she is going to >actually fall and injure herself. > >I myself have no problem in the lab, since the soles of my shoes are >rubber or plastic polymer. Sneakers work fine as well. >I manage a neuropathology laboratory which means I use paraffin embedding. >Although, I keep the floor clean, I think that the residual wax near the >embedding and sectioning stations may get spread around the rest of the >lab by my shoes. > >So far, I have put a "CAUTION" sign up on my laboratory door advising >people to excercise caution when entering the lab, especially when wearing >dress shoes, in order to at least increase awareness. > >Perhaps, I should put the laboratory floor on a regular "preventative >maintenance" schedule of cleaning and waxing to minimize the amount of wax >on the floor. >Then again, maybe I should ask the maintenance people not to put any wax >on the floor after they clean it. Perhaps it is this wax that is part of >the problem. > >Has anyone ever had this problem? How did you solve it? > >I would appreciate any advice anyone may have. > >Tim Wheelock >Harvard Brain Tissue Resource Center >McLean Hospital >617-855-3592 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sbreeden <@t> nmda.nmsu.edu Thu Sep 1 13:11:21 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: I put down the sticky mats and refer to them as PATHOLOGIST TRAPS!! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From twheelock <@t> mclean.harvard.edu Thu Sep 1 13:17:22 2005 From: twheelock <@t> mclean.harvard.edu (Tim Wheelock) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON FLOORS..a PLAN and ONE MORE QUESTION Message-ID: <3F306829.9010200@mclean.harvard.edu> Thanks so much for everyone's input. I really appreciate it. I will start by doing the following: 1) Contact Building Services and ask them strip the wax off, then strip/clean the floor, say once a month, and then to scrub-buff the most problematic areas each week. 2) Keep the warning sign up on the door. 3) Try sticky mats in an attempt to contain the paraffin around the embedding, cutting, and processing areas. 4) Contact the head of safety at McLean about the issue, as well as the head of the safety sub-committee that I sit on for suggestions. 5) Bring up the issue at our monthly staff meeting. 6) Put all this this in writing so as to make this official and "cover" myself. One more question: I have looked at the VRW on line catalogue. They advertise 2 kinds of sticky mats. "Critical Step multi-layered floor mats" and "Clean Step Multi-layered floor mats with frames" Is one better than the other for histology labs? Can you use rolling chairs on the sticky mats? Thank you, Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital Belmont, MA 617-855-3592 From petepath <@t> yahoo.com Thu Sep 1 13:45:41 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] sentinel biopsies Message-ID: <20050901184541.60268.qmail@web30406.mail.mud.yahoo.com> This is a difficult question. Our advisors in radiation here assure us the dose is quite minimal and does not require precautions. They say the exposure is similar to walking down the street ( I am guessing the street is next to a nuclear facility). We handle these as routine specimens, cut frozens and take no special percautions. Below is a consensus article from the Association of Directors of Anatomic and Surgical Pathology that covers this topic. This article recommends a variety of precautionary steps. Fitzgibbons, P, et al. Recommendations for Handling Radioactive Specimens Obtained by Sentinal Node Biopsy The American Journal of Surgical Pathology 24(11):1549 -155, 2000. I do not have a good answer to this question. I would hate to see people taking a lot of unnecessary precautions, on the other hand, I hope my other hand doesn't fall off! Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From kalschev <@t> svm.vetmed.wisc.edu Thu Sep 1 14:15:25 2005 From: kalschev <@t> svm.vetmed.wisc.edu (Vicki Kalscheur) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Hard Tissue Committee Meeting @ NSH 2005' Message-ID: <003301c5af29$81afa8e0$65d56880@kalscheur426> Dear Members and Interested Attendees: Please join us on Saturday, September 10th, 2005 from 5:30 - 6:30 p.m. in Fort Lauderdale, Florida as we gather at NSH's 31st Annual Symposium located in the Broward County Convention Center. The location/room number will be posted at the Hard Tissue Display Table. I look forward to seeing many of you! Best regards, Vicki Kalscheur, Chairperson From HornHV <@t> archildrens.org Thu Sep 1 14:36:28 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] sentinel biopsies Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE15@EMAIL.archildrens.org> In getting ready for my CAP inspection, CAP states there must be a policy for these types of specimens. My question to all of you is, who wrote your policy? Did you write it or did your Radiation Safety Officer write it? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Thursday, September 01, 2005 1:46 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] sentinel biopsies This is a difficult question. Our advisors in radiation here assure us the dose is quite minimal and does not require precautions. They say the exposure is similar to walking down the street ( I am guessing the street is next to a nuclear facility). We handle these as routine specimens, cut frozens and take no special percautions. Below is a consensus article from the Association of Directors of Anatomic and Surgical Pathology that covers this topic. This article recommends a variety of precautionary steps. Fitzgibbons, P, et al. Recommendations for Handling Radioactive Specimens Obtained by Sentinal Node Biopsy The American Journal of Surgical Pathology 24(11):1549 -155, 2000. I do not have a good answer to this question. I would hate to see people taking a lot of unnecessary precautions, on the other hand, I hope my other hand doesn't fall off! Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From dellav <@t> musc.edu Thu Sep 1 14:59:02 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Hurricane Relief Message-ID: >>> "Norman, Barbara" 09/01/05 01:15PM >>> Sounds great, NSH is working on that as we speak, I'm sure more details will follow in the next few days. Bless the American Spirit! >>> Allow me to clarify Barbara's comment and also put my two cents in. The NSH will have a computer available at the Symposium in Fort Lauderdale that will be dedicated for individuals wishing to make a donation to the Red Cross for Hurricane relief. Please keep in mind that you can do this now by going to www.redcross.org I have already done so. This would be more expedient than waiting for the opportunity to do so at the conference if you do not wish to donate to the Red Cross, you can go to http://www.fema.gov/news/newsrelease.fema?id=18473 to find a list of charities that are accepting cash donations. The NSH will NOT be accepting cash donations to be forwarded on for you , simply because you are entitled to a receipt from the charitable organization for your donation that is tax deductible. If you make a donation to NSH you will not receive the tax benefit. We (NSH) are happy to assist with information such as that contained in this message. If you are inclined to make a donation, it would probably be of greater benefit to do so sooner rather than later. for this reason and the others stated above. we have no plan to organize fund raising at the conference. I think we can have a quicker impact by acting now individually and not waiting. I don't wish to discourage the sentiments that have been expressed on histonet for those affected by the Hurricane but please keep in mind that your remarks are not likely to be seen anytime soon by those most affected. Power is expected to be out for several weeks. I know that you share my concern for our colleagues in the affected regions. Unfortunately as you know, communication is seriously hampered so it is difficult to learn if our friends and colleagues are safe. I hope that those who may have evacuated and have computer access will check in and let us know that they are safe. We have heard from Dot Kuebler, NSH Region VI Director, and Natasha Haynes, Managing Editor for the Journal of Histotechnology. I'm happy to report that they are both well and uninjured. I'll be happy to pass along news of other colleagues as it comes in. sincerely Vinnie Della Speranza President, National Society for Histotechnology From Kemlo.Rogerson <@t> elht.nhs.uk Fri Sep 2 04:37:15 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD525@elht-exch1.xelht.nhs.uk> One of our BMS's slipped on the corridor floor that sits between the 'cutting up' room and the main Lab. We have roughened strips that are supposed to prevent this but all they do is act as a reservoir. There are available special 'mats' for trapping wax but they must be cleaned regularly. Why can't people walk around in bare feet? Or wear wellies? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine Sent: 01 September 2005 17:05 To: Marshall Terry Dr,Consultant Histopathologist; Tim Wheelock; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR It may not necessarily the dress shoes. None of the shoes I wear are such but one pair tend to slip on the paraffin-coated floors more than the others. Also, we often have non-lab personnel enter the lab for a variety of reasons, for example our secretary. The new non-slip mats would probably be a big help. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, September 01, 2005 10:49 AM To: Tim Wheelock; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR Get the silly woman to wear sensible shoes:-) People who skate on thin ice cannot complain if they fall in. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Tim Wheelock [mailto:twheelock@mclean.harvard.edu] Sent: 05 August 2003 23:55 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] SLIPPING ON LAB FLOOR Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Sep 2 04:38:07 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Re: SLIPPING ON LAB FLOOR Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698F4D@elht-exch1.xelht.nhs.uk> What about dyslexics? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: 01 September 2005 19:09 To: 'Gayle Callis '; 'histonet-bounces@lists.utsouthwestern.edu '; 'Tim Wheelock '; 'Histonet@lists.utsouthwestern.edu ' Subject: RE: [Histonet] Re: SLIPPING ON LAB FLOOR Actually, you should put some kind of warning to this effect in writing and deliver it to all in the laboratory as well as put a warning to that effect on the door. This will cover you incase of a resulting law suit. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Tim Wheelock; Histonet@lists.utsouthwestern.edu Sent: 9/1/2005 12:06 PM Subject: [Histonet] Re: SLIPPING ON LAB FLOOR She is putting herself at risk for the sake of being a glamourous professional. Been there, did this in 60's - but with stilettos and very pointy toe styles, and suffered through the "slip sliding away" thing in a histology laboratory. It took a near disaster to teach an eyeopening safety awareness lesson and buying sensible safe shoes. One of the most common accidents in the work place is falling. She should use practical ( and some are not that ugly!) work related shoes, slip on style, for daily wear in office/lab, and change in a flash when she needs to be the "dressed to the nines" professional for meetings, conferences, etc. You can minimize paraffin debris by putting sticky mats (3M, sold by vendors) under your paraffin embedding, microtomy and processing areas, and in front of doorways. We put industrial rug/mats in those areas but clinical labs may have a hard time with cleaning. Our chairs roll over the mats easily, and they can be vacuumed without difficulty. At 04:55 PM 8/5/2003, you wrote: >Hi Everyone: > >A problem. >My boss, who wears dress shoes with half-size high heels (the heels have a >broad base to them, not the "stiletto" type), has been having problems >slipping on my laboratory's floor. I am really afraid that she is going to >actually fall and injure herself. > >I myself have no problem in the lab, since the soles of my shoes are >rubber or plastic polymer. Sneakers work fine as well. >I manage a neuropathology laboratory which means I use paraffin embedding. >Although, I keep the floor clean, I think that the residual wax near the >embedding and sectioning stations may get spread around the rest of the >lab by my shoes. > >So far, I have put a "CAUTION" sign up on my laboratory door advising >people to excercise caution when entering the lab, especially when wearing >dress shoes, in order to at least increase awareness. > >Perhaps, I should put the laboratory floor on a regular "preventative >maintenance" schedule of cleaning and waxing to minimize the amount of wax >on the floor. >Then again, maybe I should ask the maintenance people not to put any wax >on the floor after they clean it. Perhaps it is this wax that is part of >the problem. > >Has anyone ever had this problem? How did you solve it? > >I would appreciate any advice anyone may have. > >Tim Wheelock >Harvard Brain Tissue Resource Center >McLean Hospital >617-855-3592 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Fri Sep 2 05:06:02 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: We know why bare feet would not be a good idea but I have to know: what are wellies? -----Original Message----- From: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Friday, September 02, 2005 5:37 AM To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR One of our BMS's slipped on the corridor floor that sits between the 'cutting up' room and the main Lab. We have roughened strips that are supposed to prevent this but all they do is act as a reservoir. There are available special 'mats' for trapping wax but they must be cleaned regularly. Why can't people walk around in bare feet? Or wear wellies? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine Sent: 01 September 2005 17:05 To: Marshall Terry Dr,Consultant Histopathologist; Tim Wheelock; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR It may not necessarily the dress shoes. None of the shoes I wear are such but one pair tend to slip on the paraffin-coated floors more than the others. Also, we often have non-lab personnel enter the lab for a variety of reasons, for example our secretary. The new non-slip mats would probably be a big help. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, September 01, 2005 10:49 AM To: Tim Wheelock; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR Get the silly woman to wear sensible shoes:-) People who skate on thin ice cannot complain if they fall in. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Tim Wheelock [mailto:twheelock@mclean.harvard.edu] Sent: 05 August 2003 23:55 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] SLIPPING ON LAB FLOOR Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Sep 2 05:14:30 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD527@elht-exch1.xelht.nhs.uk> Wellington boots, rubber boots that reach to your knees; I think Wellington wore them at Trafalgar as he was worried about slipping on the deck of his ship. If meant we beat the French and they are our unofficial national emblem. It was a problem on those sailing vessels, blood and stuff made it slippy so they invented rubber boots that stopped you slipping and getting your feet wet. We all wear them in the UK, I have two pairs; one green the other black. The black ones are for gardening and things, but the green ones are for 'stepping out' in. People with horses wear green wellies as they are 'upper class'. Wellies called 'Hunters' are the 'bees knees' of wellies; you can buy them off the Net from the UK. -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: 02 September 2005 11:06 To: Rogerson Kemlo (ELHT) Pathology; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR We know why bare feet would not be a good idea but I have to know: what are wellies? -----Original Message----- From: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Friday, September 02, 2005 5:37 AM To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR One of our BMS's slipped on the corridor floor that sits between the 'cutting up' room and the main Lab. We have roughened strips that are supposed to prevent this but all they do is act as a reservoir. There are available special 'mats' for trapping wax but they must be cleaned regularly. Why can't people walk around in bare feet? Or wear wellies? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine Sent: 01 September 2005 17:05 To: Marshall Terry Dr,Consultant Histopathologist; Tim Wheelock; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR It may not necessarily the dress shoes. None of the shoes I wear are such but one pair tend to slip on the paraffin-coated floors more than the others. Also, we often have non-lab personnel enter the lab for a variety of reasons, for example our secretary. The new non-slip mats would probably be a big help. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, September 01, 2005 10:49 AM To: Tim Wheelock; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR Get the silly woman to wear sensible shoes:-) People who skate on thin ice cannot complain if they fall in. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Tim Wheelock [mailto:twheelock@mclean.harvard.edu] Sent: 05 August 2003 23:55 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] SLIPPING ON LAB FLOOR Hi Everyone: A problem. My boss, who wears dress shoes with half-size high heels (the heels have a broad base to them, not the "stiletto" type), has been having problems slipping on my laboratory's floor. I am really afraid that she is going to actually fall and injure herself. I myself have no problem in the lab, since the soles of my shoes are rubber or plastic polymer. Sneakers work fine as well. I manage a neuropathology laboratory which means I use paraffin embedding. Although, I keep the floor clean, I think that the residual wax near the embedding and sectioning stations may get spread around the rest of the lab by my shoes. So far, I have put a "CAUTION" sign up on my laboratory door advising people to excercise caution when entering the lab, especially when wearing dress shoes, in order to at least increase awareness. Perhaps, I should put the laboratory floor on a regular "preventative maintenance" schedule of cleaning and waxing to minimize the amount of wax on the floor. Then again, maybe I should ask the maintenance people not to put any wax on the floor after they clean it. Perhaps it is this wax that is part of the problem. Has anyone ever had this problem? How did you solve it? I would appreciate any advice anyone may have. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital 617-855-3592 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anicoll <@t> cellpath.co.uk Fri Sep 2 06:07:34 2005 From: anicoll <@t> cellpath.co.uk (Alastair Nicoll) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: <0F42D9ABA325D811BBE300902712436D84F3D3@cellmail.cellpath.co.uk> Classic reply A small boy we were babysitting last weekend loves his yellow wellies. He wore nothing else on his feet all weekend. Ali Nicoll -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: 02 September 2005 11:15 To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR Wellington boots, rubber boots that reach to your knees; I think Wellington wore them at Trafalgar as he was worried about slipping on the deck of his ship. If meant we beat the French and they are our unofficial national emblem. It was a problem on those sailing vessels, blood and stuff made it slippy so they invented rubber boots that stopped you slipping and getting your feet wet. We all wear them in the UK, I have two pairs; one green the other black. The black ones are for gardening and things, but the green ones are for 'stepping out' in. People with horses wear green wellies as they are 'upper class'. Wellies called 'Hunters' are the 'bees knees' of wellies; you can buy them off the Net from the UK. -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: 02 September 2005 11:06 To: Rogerson Kemlo (ELHT) Pathology; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR We know why bare feet would not be a good idea but I have to know: what are wellies? From Tbarnhart <@t> primecare.org Fri Sep 2 06:56:46 2005 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] sentinel biopsies Message-ID: <1779904B5E82D511914C00D0B793339205BFD9FA@exchangent> It was a team effort. The CAP guidelines require the Radiation Safety Officer to sign off on your policy. -----Original Message----- From: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: Thursday, September 01, 2005 2:36 PM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] sentinel biopsies In getting ready for my CAP inspection, CAP states there must be a policy for these types of specimens. My question to all of you is, who wrote your policy? Did you write it or did your Radiation Safety Officer write it? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Thursday, September 01, 2005 1:46 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] sentinel biopsies This is a difficult question. Our advisors in radiation here assure us the dose is quite minimal and does not require precautions. They say the exposure is similar to walking down the street ( I am guessing the street is next to a nuclear facility). We handle these as routine specimens, cut frozens and take no special percautions. Below is a consensus article from the Association of Directors of Anatomic and Surgical Pathology that covers this topic. This article recommends a variety of precautionary steps. Fitzgibbons, P, et al. Recommendations for Handling Radioactive Specimens Obtained by Sentinal Node Biopsy The American Journal of Surgical Pathology 24(11):1549 -155, 2000. I do not have a good answer to this question. I would hate to see people taking a lot of unnecessary precautions, on the other hand, I hope my other hand doesn't fall off! Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From Kemlo.Rogerson <@t> elht.nhs.uk Fri Sep 2 08:28:14 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698F53@elht-exch1.xelht.nhs.uk> Even in bed? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Alastair Nicoll Sent: 02 September 2005 12:08 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR Classic reply A small boy we were babysitting last weekend loves his yellow wellies. He wore nothing else on his feet all weekend. Ali Nicoll -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: 02 September 2005 11:15 To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR Wellington boots, rubber boots that reach to your knees; I think Wellington wore them at Trafalgar as he was worried about slipping on the deck of his ship. If meant we beat the French and they are our unofficial national emblem. It was a problem on those sailing vessels, blood and stuff made it slippy so they invented rubber boots that stopped you slipping and getting your feet wet. We all wear them in the UK, I have two pairs; one green the other black. The black ones are for gardening and things, but the green ones are for 'stepping out' in. People with horses wear green wellies as they are 'upper class'. Wellies called 'Hunters' are the 'bees knees' of wellies; you can buy them off the Net from the UK. -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: 02 September 2005 11:06 To: Rogerson Kemlo (ELHT) Pathology; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] SLIPPING ON LAB FLOOR We know why bare feet would not be a good idea but I have to know: what are wellies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hymclab <@t> hyhc.com Fri Sep 2 09:20:16 2005 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] sentinel biopsies Message-ID: I wrote my own policy but referenced that I received the radiation information from the Hospital Radiation Safety officer. We do the same as Dr. Peters here. The Radiation Safety officer (which is a Radiologist here) said we were in no danger. The only thing he suggested and I have it in my procedure is to have any pregnant histotechs avoid handling the fresh sentinel nodes just as a precautionary measure. Hope this helps, Dawn -----Original Message----- From: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: Thursday, September 01, 2005 2:36 PM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] sentinel biopsies In getting ready for my CAP inspection, CAP states there must be a policy for these types of specimens. My question to all of you is, who wrote your policy? Did you write it or did your Radiation Safety Officer write it? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Thursday, September 01, 2005 1:46 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] sentinel biopsies This is a difficult question. Our advisors in radiation here assure us the dose is quite minimal and does not require precautions. They say the exposure is similar to walking down the street ( I am guessing the street is next to a nuclear facility). We handle these as routine specimens, cut frozens and take no special percautions. Below is a consensus article from the Association of Directors of Anatomic and Surgical Pathology that covers this topic. This article recommends a variety of precautionary steps. Fitzgibbons, P, et al. Recommendations for Handling Radioactive Specimens Obtained by Sentinal Node Biopsy The American Journal of Surgical Pathology 24(11):1549 -155, 2000. I do not have a good answer to this question. I would hate to see people taking a lot of unnecessary precautions, on the other hand, I hope my other hand doesn't fall off! Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Fri Sep 2 10:26:23 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Re: SLIPPING ON LAB FLOOR Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB43C5@fh2k093.fhmis.net> Post it on the window and they'll see it on the other side!! -----Original Message----- From: Rogerson Kemlo (ELHT) Pathology To: Bonner, Janet; Gayle Callis ; histonet-bounces@lists.utsouthwestern.edu; Tim Wheelock ; Histonet@lists.utsouthwestern.edu Sent: 9/2/2005 5:38 AM Subject: RE: [Histonet] Re: SLIPPING ON LAB FLOOR What about dyslexics? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: 01 September 2005 19:09 To: 'Gayle Callis '; 'histonet-bounces@lists.utsouthwestern.edu '; 'Tim Wheelock '; 'Histonet@lists.utsouthwestern.edu ' Subject: RE: [Histonet] Re: SLIPPING ON LAB FLOOR Actually, you should put some kind of warning to this effect in writing and deliver it to all in the laboratory as well as put a warning to that effect on the door. This will cover you incase of a resulting law suit. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Tim Wheelock; Histonet@lists.utsouthwestern.edu Sent: 9/1/2005 12:06 PM Subject: [Histonet] Re: SLIPPING ON LAB FLOOR She is putting herself at risk for the sake of being a glamourous professional. Been there, did this in 60's - but with stilettos and very pointy toe styles, and suffered through the "slip sliding away" thing in a histology laboratory. It took a near disaster to teach an eyeopening safety awareness lesson and buying sensible safe shoes. One of the most common accidents in the work place is falling. She should use practical ( and some are not that ugly!) work related shoes, slip on style, for daily wear in office/lab, and change in a flash when she needs to be the "dressed to the nines" professional for meetings, conferences, etc. You can minimize paraffin debris by putting sticky mats (3M, sold by vendors) under your paraffin embedding, microtomy and processing areas, and in front of doorways. We put industrial rug/mats in those areas but clinical labs may have a hard time with cleaning. Our chairs roll over the mats easily, and they can be vacuumed without difficulty. At 04:55 PM 8/5/2003, you wrote: >Hi Everyone: > >A problem. >My boss, who wears dress shoes with half-size high heels (the heels have a >broad base to them, not the "stiletto" type), has been having problems >slipping on my laboratory's floor. I am really afraid that she is going to >actually fall and injure herself. > >I myself have no problem in the lab, since the soles of my shoes are >rubber or plastic polymer. Sneakers work fine as well. >I manage a neuropathology laboratory which means I use paraffin embedding. >Although, I keep the floor clean, I think that the residual wax near the >embedding and sectioning stations may get spread around the rest of the >lab by my shoes. > >So far, I have put a "CAUTION" sign up on my laboratory door advising >people to excercise caution when entering the lab, especially when wearing >dress shoes, in order to at least increase awareness. > >Perhaps, I should put the laboratory floor on a regular "preventative >maintenance" schedule of cleaning and waxing to minimize the amount of wax >on the floor. >Then again, maybe I should ask the maintenance people not to put any wax >on the floor after they clean it. Perhaps it is this wax that is part of >the problem. > >Has anyone ever had this problem? How did you solve it? > >I would appreciate any advice anyone may have. > >Tim Wheelock >Harvard Brain Tissue Resource Center >McLean Hospital >617-855-3592 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From linhines <@t> yahoo.com Fri Sep 2 11:18:36 2005 From: linhines <@t> yahoo.com (Linda Hines) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] RE: Histo-Techs ON CALL Message-ID: <20050902161836.50691.qmail@web33011.mail.mud.yahoo.com> Hello, Histonetters I would like to inform everyone about a new company called Histo-Techs on call. We do private histology contracting work for pathology laboratories that fine themselves TOO SHORT STAFFED!!! or in a bind. We are ASCP REGISTERED TECHNICIANS with routine histology, special stains, immuno's, and MOHS experience.If you need a histologist to fill-in for unexpected illnesses, vacations, etc. Call Us @ (480) 252-0050 or (602) 441-3268. We are located Phoenix, AZ. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From John.Auld <@t> whnt.nhs.uk Fri Sep 2 11:41:51 2005 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Re: Slipping on floors Message-ID: My understanding is that in the UK you should not use wheeled chairs on non carpeted floors. We recently had new flooring installed in the histo lab and went for non slip flooring similar to that used in swimming pool areas. It's sooo good much much better than the old wax impregnated lino that had been down for years.. The cleaners regularly stipped and buffed the old stuff but even early in the day before work got underway it was still slippy. Get your employer to bite the bullet and get real non slip stuff down before you get sued you know it makes sense Have a good weekend and our thoughts are with those suffering from the effects of Hurricane Katrina John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 From: Tim Wheelock Subject: [Histonet] SLIPPING ON FLOORS..a PLAN and ONE MORE QUESTION To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3F306829.9010200@mclean.harvard.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Thanks so much for everyone's input. I really appreciate it. I will start by doing the following: 1) Contact Building Services and ask them strip the wax off, then strip/clean the floor, say once a month, and then to scrub-buff the most problematic areas each week. 2) Keep the warning sign up on the door. 3) Try sticky mats in an attempt to contain the paraffin around the embedding, cutting, and processing areas. 4) Contact the head of safety at McLean about the issue, as well as the head of the safety sub-committee that I sit on for suggestions. 5) Bring up the issue at our monthly staff meeting. 6) Put all this this in writing so as to make this official and "cover" myself. One more question: I have looked at the VRW on line catalogue. They advertise 2 kinds of sticky mats. "Critical Step multi-layered floor mats" and "Clean Step Multi-layered floor mats with frames" Is one better than the other for histology labs? Can you use rolling chairs on the sticky mats? Thank you, From tahseen <@t> brain.net.pk Fri Sep 2 06:04:37 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Protocol of Immunofluorescence Techniques. Message-ID: <005b01c5afae$1dbb1f60$972bfea9@m7c0y4> Hi All, 1 Does anyone here can share me a practical protocol of Immunofluorescence Techniques on frozen sections? 2 Has anyone got a recipe for Zeus Scientific tissue Fixative? Muhammad Tahseen Histology Supervisor, SKMCH & RC Lahore Pakistan. From leswes <@t> shaw.ca Fri Sep 2 13:39:28 2005 From: leswes <@t> shaw.ca (Lesley Weston) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR In-Reply-To: <53A22BBB01D6184EBF7A4DC18A021E9E5AD527@elht-exch1.xelht.nhs.uk> Message-ID: Wasn't that Nelson? I thought the Duke of Wellington invented rubber riding boots, which made riders' feet so dry and comfortable that everybody started wearing them for everything. They would still slip on waxy floors, though. Lesley Weston. > From: "Rogerson Kemlo (ELHT) Pathology" > Date: Fri, 02 Sep 2005 11:14:30 +0100 > To: "Bartlett, Jeanine" , histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] SLIPPING ON LAB FLOOR > > Wellington boots, rubber boots that reach to your knees; I think > Wellington wore them at Trafalgar as he was worried about slipping on > the deck of his ship. If meant we beat the French and they are our > unofficial national emblem. > > It was a problem on those sailing vessels, blood and stuff made it > slippy so they invented rubber boots that stopped you slipping and > getting your feet wet. We all wear them in the UK, I have two pairs; one > green the other black. The black ones are for gardening and things, but > the green ones are for 'stepping out' in. People with horses wear green > wellies as they are 'upper class'. Wellies called 'Hunters' are the > 'bees knees' of wellies; you can buy them off the Net from the UK. > > -----Original Message----- > From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] > Sent: 02 September 2005 11:06 > To: Rogerson Kemlo (ELHT) Pathology; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] SLIPPING ON LAB FLOOR > > We know why bare feet would not be a good idea but I have to know: what > are wellies? > > -----Original Message----- > From: Rogerson Kemlo (ELHT) Pathology > [mailto:Kemlo.Rogerson@elht.nhs.uk] > Sent: Friday, September 02, 2005 5:37 AM > To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] SLIPPING ON LAB FLOOR > > One of our BMS's slipped on the corridor floor that sits between the > 'cutting up' room and the main Lab. We have roughened strips that are > supposed to prevent this but all they do is act as a reservoir. There > are available special 'mats' for trapping wax but they must be cleaned > regularly. > > Why can't people walk around in bare feet? Or wear wellies? From relia1 <@t> earthlink.net Fri Sep 2 14:30:55 2005 From: relia1 <@t> earthlink.net (Pam Barker) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] RELIA Histology Job Opportunity Update 9/2/05 - From Pam Barker Message-ID: Hello Histonetters, I just wanted to drop you a line and tell you about my current histology openings. All the positions I represent are permanent positions with great companies who offer excellent compensation, benefits and relocation/sign on bonuses. Wherever you want to go, wherever you want to stay if you are looking for a new opportunity I can help. I offer over 20 years of recruiting experience and a recruiting practice solely dedicated to permanent placement in the Histology profession. In addition to representing you to the positions you are interested in with my clients I offer to you FREE of Charge: ? Assistance with updating or creating your resume ? Tips on interviewing ? Encouragement and assistance during the course of your job search ? Complete Confidentiality ( I will only represent you to jobs you tell me you are interested in looking into) Here is a list of my newest openings 1. Senior Research Specialist Immunohistochemistry - California 2. Histo Tech (2 positions) Clinical Setting - Southeastern Georgia 3. Histo Tech (2 positions) Clinical Setting - Southwestern Florida 4. Technical Support Specialist - Research Setting - Northern California Here is a list of my current openings: HISTOLOGY MANAGEMENT: Anatomic Pathology Manager ? South Carolina Anatomic Pathology Manager ? Boston, MA Histology Supervisor ? Boston, MA Histology Supervisor ? Pennsylvania Histology Supervisor ? South Florida HISTOTECHNICIAN/TECHNOLOGIST: Histo Tech (clinical setting) ? Northern, CA Histo Tech (research setting) ? Northern, CA Histo Tech (clinical setting) ? Central Florida Histo Tech (clinical setting) - South Florida Histo Tech (clinical setting) ? Boston, MA MOHS/Histo Tech ? South Florida Histo Tech (clinical setting) - Minnesota Histo Tech (clinical setting) ? Alaska Histo Tech (Night shift) - Colorado Histo Tech (clinical setting) Indiana - (close to Chicago) If you or any of your friends would like more information on any of these positions. Or if you would like to discuss opportunities in other areas or future job searches please contact me. I would be more than happy to assist you. You can reach me at 866-607-3542 or relia1@earthlink.net I work with facilities nationwide and I will keep your resume confidential. I will only represent you to jobs you tell me you are interested in looking into. Remember It never hurts to keep an eye open even if you are happy in your present job. Have a Safe and Happy Labor Day Weekend! Thanks! Pam ? 866-607-3542 (866-60RELIA) Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net From froyer <@t> bitstream.net Fri Sep 2 14:59:34 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Re: Slipping on floors Message-ID: Ford M. Royer, MT(ASCP) Sales Manager - Histology Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Minneapolis, MN 55427-3601 phone: 888-790-9686 or 763-542-8725 fax: 763-546-4830 email: web: -----Original Message----- From: Ford Royer [mailto:froyer@bitstream.net] Sent: Friday, September 02, 2005 2:58 PM To: John Auld Subject: RE: [Histonet] Re: Slipping on floors WOT!! No wheeled chair races through the corridors of the Laboratory in the U.K.?! How sad... The last Lab I worked in, I was considered the Parnelli Jones of four-wheeled extreme chair racing. Only had one career mishap... took out the Chief of Pathology when he stepped out into the hallway from his office. It wasn't my fault. He could have avoided the whole thing if those 25, fully loaded, slide trays he was carrying had not blocked his view. Everyone knows you should not carry more than 2 trays at a time. ~ Ford ;-) Ford M. Royer, MT(ASCP) Sales Manager - Histology Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Minneapolis, MN 55427-3601 phone: 888-790-9686 or 763-542-8725 fax: 763-546-4830 email: web: -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of John Auld Sent: Friday, September 02, 2005 11:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Slipping on floors My understanding is that in the UK you should not use wheeled chairs on non carpeted floors. We recently had new flooring installed in the histo lab and went for non slip flooring similar to that used in swimming pool areas. It's sooo good much much better than the old wax impregnated lino that had been down for years.. The cleaners regularly stipped and buffed the old stuff but even early in the day before work got underway it was still slippy. Get your employer to bite the bullet and get real non slip stuff down before you get sued you know it makes sense Have a good weekend and our thoughts are with those suffering from the effects of Hurricane Katrina John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 From: Tim Wheelock Subject: [Histonet] SLIPPING ON FLOORS..a PLAN and ONE MORE QUESTION To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3F306829.9010200@mclean.harvard.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Thanks so much for everyone's input. I really appreciate it. I will start by doing the following: 1) Contact Building Services and ask them strip the wax off, then strip/clean the floor, say once a month, and then to scrub-buff the most problematic areas each week. 2) Keep the warning sign up on the door. 3) Try sticky mats in an attempt to contain the paraffin around the embedding, cutting, and processing areas. 4) Contact the head of safety at McLean about the issue, as well as the head of the safety sub-committee that I sit on for suggestions. 5) Bring up the issue at our monthly staff meeting. 6) Put all this this in writing so as to make this official and "cover" myself. One more question: I have looked at the VRW on line catalogue. They advertise 2 kinds of sticky mats. "Critical Step multi-layered floor mats" and "Clean Step Multi-layered floor mats with frames" Is one better than the other for histology labs? Can you use rolling chairs on the sticky mats? Thank you, _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Fri Sep 2 17:45:38 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Confocal systems - who's the best? In-Reply-To: <005b01c5afae$1dbb1f60$972bfea9@m7c0y4> References: <005b01c5afae$1dbb1f60$972bfea9@m7c0y4> Message-ID: Hi All, We are looking to purchase a confocal microscope for inverted microscopy with attachments for live-cell imaging. Can everyone please give me a run down of the pluses and minuses of various systems and companies (Nikon, Olympus, Zeiss, Leica) and overall what we need to look for. Which is the best company???? THANK YOU! Andrea -- From gliuygao <@t> hotmail.com Sat Sep 3 10:47:02 2005 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Cut brain Message-ID: I have a problem cut flat section on 8 um cerebellum. Any tips? Yan From abright <@t> brightinstruments.com Sat Sep 3 17:08:33 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Cut brain Message-ID: Yan, Yes many, please can I have more detail of your problem. Is this frozen tissue or wax impregnated? Alan Bright Bright Instrument Company England -----Original Message----- From: yan gao [mailto:gliuygao@hotmail.com] Sent: Sat 03/09/2005 16:47 To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Cut brain I have a problem cut flat section on 8 um cerebellum. Any tips? Yan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dbpiontek <@t> hotmail.com Sat Sep 3 20:57:54 2005 From: dbpiontek <@t> hotmail.com (Denise Piontek) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Job Oppurtunity in MA Message-ID: Growing Contract Research Organization specializing in biocompatibility and toxicity studies for the medical device and pharmaceutical industries is in need of a full-time, experienced histology technician or histotechnologist. The person, as a team player, will work with other histology technicians, veterinarian, pathologist, Study Directors, and animal technicians to produce quality work. Responsibilities will include: * Trimming / grossing of tissues and organs harvested at necropsy * Embedding and processing of tissues * Microtomy * Routine H&E staining, special stains as required and special techniques as attained * Maintain and/or create GLP documentation as necessary, including relative SOPs and histology records * ISO regulated facility * Assist in necropsy procedures as necessary * Maintain wet tissue archive * Data evaluations of various project aspects * Great oppurtunity to increase or use anatomy skills Experience and a high school diploma (or GED) are required. A college degree Associates or Bachelors), research experience, certification by the ASCP, and/or experience with animal tissues are strongly preferred. Willingness to learn, attention to detail, proficiency with computers, good organizational skills, and well-developed time management skills are also desired. Salary is commiserate with experience. Competitive benefits. Position is M-F 7AM to 4 PM or 8AM to 5 PM. We are located just outside of Boston, MA. Please submit resumes to: Human Resources Toxikon Corporation 15 Wiggins Ave. Bedford, MA 01730 [1]hr@toxikon.com Also, visit us on the web (apply online) at [2]www.toxikon.com _________________________________________________________________ [3]Find e-mail and documents on your PC instantly with the new MSN Search ToolbarFREE! References 1. mailto:hr@toxikon.com?subject=Histology%20Technician%2FHistotechnologist%20Position 2. http://www.toxikon.com/ 3. http://g.msn.com/8HMAENUS/2734??PS=47575 From Kemlo.Rogerson <@t> elht.nhs.uk Mon Sep 5 03:07:30 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] SLIPPING ON LAB FLOOR Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698F56@elht-exch1.xelht.nhs.uk> Dang, you spoiled a good story! Could figure out why Wellington would wear boots that all never mind. My Weliies have never slipped on waxy floors, wearing them now. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lesley Weston Sent: 02 September 2005 19:39 To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] SLIPPING ON LAB FLOOR Wasn't that Nelson? I thought the Duke of Wellington invented rubber riding boots, which made riders' feet so dry and comfortable that everybody started wearing them for everything. They would still slip on waxy floors, though. Lesley Weston. > From: "Rogerson Kemlo (ELHT) Pathology" > Date: Fri, 02 Sep 2005 11:14:30 +0100 > To: "Bartlett, Jeanine" , histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] SLIPPING ON LAB FLOOR > > Wellington boots, rubber boots that reach to your knees; I think > Wellington wore them at Trafalgar as he was worried about slipping on > the deck of his ship. If meant we beat the French and they are our > unofficial national emblem. > > It was a problem on those sailing vessels, blood and stuff made it > slippy so they invented rubber boots that stopped you slipping and > getting your feet wet. We all wear them in the UK, I have two pairs; one > green the other black. The black ones are for gardening and things, but > the green ones are for 'stepping out' in. People with horses wear green > wellies as they are 'upper class'. Wellies called 'Hunters' are the > 'bees knees' of wellies; you can buy them off the Net from the UK. > > -----Original Message----- > From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] > Sent: 02 September 2005 11:06 > To: Rogerson Kemlo (ELHT) Pathology; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] SLIPPING ON LAB FLOOR > > We know why bare feet would not be a good idea but I have to know: what > are wellies? > > -----Original Message----- > From: Rogerson Kemlo (ELHT) Pathology > [mailto:Kemlo.Rogerson@elht.nhs.uk] > Sent: Friday, September 02, 2005 5:37 AM > To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] SLIPPING ON LAB FLOOR > > One of our BMS's slipped on the corridor floor that sits between the > 'cutting up' room and the main Lab. We have roughened strips that are > supposed to prevent this but all they do is act as a reservoir. There > are available special 'mats' for trapping wax but they must be cleaned > regularly. > > Why can't people walk around in bare feet? Or wear wellies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ree3 <@t> leicester.ac.uk Mon Sep 5 11:26:34 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] beta-2- microglobulin Message-ID: Looking for an antibody to the above that works, as ever, on paraffin processed mouse tissues... Many thanks Richard Edwards MRC TOX UNIT LEICESTER...U.K.... From Debbiejsiena <@t> aol.com Mon Sep 5 12:39:13 2005 From: Debbiejsiena <@t> aol.com (Debbiejsiena@aol.com) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Help wanted (Lab Aid) Dallas, Texas Message-ID: <81.2f4fbfe7.304ddcc1@aol.com> Good morning everyone We at Tissue Techniques in Dallas, Texas are asking for your help. Our lab is looking for someone who would be a good lab aid. We are willing to train that someone who is the correct person. If anyone knows of that person please let us hear from you. Thanks again for all of your help. Sincerely; Debbie Siena HT (ASCP) QIHC From jstaruk <@t> masshistology.com Mon Sep 5 17:42:10 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Job opening (with free room and board and transportation) In-Reply-To: <81.2f4fbfe7.304ddcc1@aol.com> Message-ID: <49k0vg$65vs26@mxip29a.cluster1.charter.net> I'm not sure if this message will reach those in need, but Mass Histology Service is offering a position within our company (histologist or lab aid) to a qualified person who has been left homeless and jobless by the recent hurricane. I have a comfortable spare room at my house to keep them warm and a stocked kitchen to feed them. They can also ride in with me as well as ride back home with me (although I usually work 12 hour days). They will be welcomed to stay in my house until they get their feet back on the ground. Unfortunately, I cannot offer this opportunity to an entire family, only a single, qualified person. If anyone on this listserver knows of someone in need, please have them contact me. Mass Histology Service has also made a considerable donation to the American Red Cross and we hope others on this server will follow to help those less fortunate than us. Jim ____________________ Jim Staruk www.masshistology.com From ree3 <@t> leicester.ac.uk Tue Sep 6 03:33:17 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] RE: beta-2- microglobulin Message-ID: Looking for an antibody to the above that works, as ever, on paraffin processed mouse tissues... Many thanks Richard Edwards MRC TOX UNIT LEICESTER...U.K.... From christelle.gerard <@t> novartis.com Tue Sep 6 08:07:20 2005 From: christelle.gerard <@t> novartis.com (christelle.gerard@novartis.com) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] anti VEGF on mouse tissue Message-ID: Hello, I already tested differents antibodies from Santa Cruz to detect VEGF on mouse tissues fixed in formalin and embedded in paraffin. There was no result. Have you any experience in this IHC? Thanks in advance for any informations, references or protocoles. Christelle From christelle.gerard <@t> novartis.com Tue Sep 6 08:09:55 2005 From: christelle.gerard <@t> novartis.com (christelle.gerard@novartis.com) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] anti Cy5 Message-ID: Hello, I m looking for antibody against Cy5.5. Did someone already use? Have you any informations? Thanks in davance. Christelle From billingconsultants <@t> yahoo.com Tue Sep 6 08:21:34 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Histotech Position: Athens Georgia Message-ID: <20050906132134.55688.qmail@web54204.mail.yahoo.com> Hi Everyone, There is a position open in Athens, Georgia for a histotech to fill in from November through January. If you're interested, please call me at 706-546-0200. Thank you, Louri Roberts __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From YinongChen <@t> texashealth.org Tue Sep 6 08:32:55 2005 From: YinongChen <@t> texashealth.org (Chen, Yinong) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] subscribe Message-ID: Please subscribe The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From John.MacDougall <@t> serono.com Tue Sep 6 09:57:06 2005 From: John.MacDougall <@t> serono.com (John.MacDougall@serono.com) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Frequency of alcohol/xylene/paraffin replacement in tissue processor Message-ID: Dear histonetters I'm new to the Shandon Citadel Tissue Processor and was wondering about the frequency at which I should replace the alcohols, xylene and paraffin... Is this based on volume of use, length of time or (likely) both.... Thanks John John MacDougall, Ph.D. Senior Principal Investigator - Biotherapeutic Discovery Research and Pharmaceutical Development Serono, Inc. One Technology Place Rockland, MA 02370 U.S.A. Voice - 1-781-681-2872 Fax - 1-781-681-2910 From JWEEMS <@t> sjha.org Tue Sep 6 10:19:42 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] sentinel biopsies Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01304BE0@sjhaexc02.sjha.org> We went through this several years ago and by inspection two years ago were no longer treating them any differently. I removed the procedure from the manual but was sited for not having one. For those of you not treating them differently, what kind of procedure do you have? I need to be correct for inspection coming up in October! Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of hymclab Sent: Friday, September 02, 2005 10:20 AM To: 'Horn, Hazel V'; Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] sentinel biopsies I wrote my own policy but referenced that I received the radiation information from the Hospital Radiation Safety officer. We do the same as Dr. Peters here. The Radiation Safety officer (which is a Radiologist here) said we were in no danger. The only thing he suggested and I have it in my procedure is to have any pregnant histotechs avoid handling the fresh sentinel nodes just as a precautionary measure. Hope this helps, Dawn -----Original Message----- From: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: Thursday, September 01, 2005 2:36 PM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] sentinel biopsies In getting ready for my CAP inspection, CAP states there must be a policy for these types of specimens. My question to all of you is, who wrote your policy? Did you write it or did your Radiation Safety Officer write it? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Thursday, September 01, 2005 1:46 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] sentinel biopsies This is a difficult question. Our advisors in radiation here assure us the dose is quite minimal and does not require precautions. They say the exposure is similar to walking down the street ( I am guessing the street is next to a nuclear facility). We handle these as routine specimens, cut frozens and take no special percautions. Below is a consensus article from the Association of Directors of Anatomic and Surgical Pathology that covers this topic. This article recommends a variety of precautionary steps. Fitzgibbons, P, et al. Recommendations for Handling Radioactive Specimens Obtained by Sentinal Node Biopsy The American Journal of Surgical Pathology 24(11):1549 -155, 2000. I do not have a good answer to this question. I would hate to see people taking a lot of unnecessary precautions, on the other hand, I hope my other hand doesn't fall off! Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From pmarcum <@t> vet.upenn.edu Tue Sep 6 10:44:28 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Further information of animal rescue and assistance Message-ID: <6.1.1.1.2.20050906113419.0199a138@mail.vet.upenn.edu> Good Morning All, The following attachment is from the Dean of LSU Veterinary School in Baton Rogue. I know we have so many evacuees to worry about and the animals can be forgotten in the wake of the disaster. I have spoken to a person in Houston (she can let you know what she did if she wishes) and I asked her about the animals that were coming in with the evacuees. She says they are housed in a kennel type area near the complex and cared for by local veterinarians and volunteers. They will also need the same types of materials listed below. I am not sure what has been set up in Texas but I sure if you are local and have experience in the Vet field or can help please try to contact some one and help. I again, hope no one is offended by this posting however we domesticate these animals and now they are suffering right along with the owners. A MESSAGE FROM THE DEAN We at the School of Veterinary Medicine, like everyone else in the country, are overwhelmed by what we have seen on the news in the wake of Hurricane Katrina. In an effort to help our colleagues and fellow citizens, the School is working with the Louisiana Veterinary Medical Association, the Louisiana Animal Control Association, and the Louisiana SPCA to provide shelter and care for those pets that have traveled with their owners from the flooded areas and animals that have been rescued from those areas. The School's faculty, staff and students are volunteering their time at the Parker Coliseum on the LSU campus and the Lamar-Dixon Expo Center in Gonzales, La., where animals are being sheltered. This is not something that will end in a few days. The School will require on-going support from the community. We desperately need volunteers, especially veterinarians and veterinary technicians, to help us in this effort. Non-veterinary volunteers are also welcome, though only those people that have been vaccinated for rabies will be able to work directly with the animals. The animal shelters are in dire need of large crates and cages. The volunteers also need ice, beverages and food. Other animal supplies, such as food, cat litter, pooper scoopers, vaccines, antibiotics, bandages, and catheters are also welcome and needed. Monetary donations can be made to the Louisiana Veterinary Medical Association by calling 1-800-524-2996 or 225-928-5862. You can also download a donation form at the LVMA website at www.lvma.org, or send a check or money order made payable to the Dr. Walter J. Ernst, Jr. Veterinary Memorial Foundation, 8550 United Plaza Blvd., Suite 1001, Baton Rouge, LA 70809. It is during times of adversity and tragedy that people must come together. The School of Veterinary Medicine, along with state and local animal organizations, wants to do our part by caring for as many animals as possible. Please help us in our efforts. Veterinarians and veterinary technicians who want to volunteer assistance can contact Dr. David Senior at the School of Veterinary Medicine at 225-578-9551 or dsenior@vetmed.lsu.edu. Non-veterinary volunteers can contact the School of Veterinary Medicine at 225-578-9900. Thank you for your prayers and support. Sincerely, Michael G. Groves, DVM Dean School of Veterinary Medicine Louisiana State University Baton Rouge, LA 70803 Phone: (225) 578-9900/Fax: (225) 578-9916/Web: www.vetmed.lsu.edu --- Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From pmarcum <@t> vet.upenn.edu Tue Sep 6 11:41:41 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Apology about earlier post Message-ID: <6.1.1.1.2.20050906123457.01a10d30@mail.vet.upenn.edu> Please note the states of Alabama and Mississippi also have animals in need and I am sure they have areas that can use the same materials and supplies for pets and some farm animals that have been rescued. I did not mean to ignore them I have family in Louisiana and I get information from there and Houston often so that area tends to be on my mind more. Please visit sites for Vet Schools or SPCA in both states as I am sure they have ways to assist directly for their states. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From pruegg <@t> ihctech.net Tue Sep 6 12:12:08 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] viability Message-ID: <200509061712.j86HC0fj007153@chip.viawest.net> Has anyone use Tetrazolium Salt salt to measure viability in live tissue, not touch preps or frozen sections, fresh tissue pieces as they are removed at necropsy? We are wondering if you put the salt on the live tissue if it would turn blue? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From pruegg <@t> ihctech.net Tue Sep 6 12:16:35 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] anti VEGF on mouse tissue In-Reply-To: Message-ID: <200509061716.j86HGRfj008471@chip.viawest.net> I use monoclonal vegf from Santa Cruz, there is only one, at 1:200 with HIER (I use steam for 20 min. with citrate buffer ph6), over night incubation of the primary at 4dc and labelled polymer hrp detection for 60 min. at rt, DAB 10 min. I have also found it necessary to use h202 with methanol to quench endogenous peroxidase. Oops, this is not going to work on you mouse tissue, guess you will have to try one of the polyclonal vegf's or a ms on ms detection. I use monoclonal vegf on human, rabbit, sheep and rat tissue, not mouse. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of christelle.gerard@novartis.com Sent: Tuesday, September 06, 2005 6:07 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] anti VEGF on mouse tissue Hello, I already tested differents antibodies from Santa Cruz to detect VEGF on mouse tissues fixed in formalin and embedded in paraffin. There was no result. Have you any experience in this IHC? Thanks in advance for any informations, references or protocoles. Christelle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Tue Sep 6 15:54:03 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] CD26 on FFPE Message-ID: <355C35514FEAC9458F75947F5270974D0795F0@usctmx1103.merck.com> Anybody have success with CD26 IHC on FFPE sections? Care to share your antibody source? Thanks, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From Eric <@t> ategra.com Tue Sep 6 15:41:57 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Immediate Job Opportunities For HistoTechs (perm and contract) Message-ID: Histonetters - I have temporary travel assignments and permanent HistoTech openings available for you right now throughout the Southeast and the rest of the country! If you are currently available, please contact me ASAP at (800) 466-9919, ext. 223. Our clients are now hiring and need to fill these positions ASAP. Thanks for your time/attention... Eric Dye (800) 466-9919, ext. 223 PS - If you know anyone else who is currently looking, I would appreciate it if you could email me there phone number/email address. (Your friend would also appreciate it too!) PSS - If YOU are interested in any of these opportunities, please call me ASAP @ (800) 466 9919 x223. To speed things up, please send me a copy of your resume, (if you haven't already done so) - There is absolutely N O C O S T to you whatsoever! Our services are entirely paid for by the hiring companies. Eric Dye (800) 466-9919, ext. 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792 (800)466-9919 ext 231 FAX: (407) 671-6075 eric@ategra.com To Learn More About Ategra: http://www.ategra.com ------------------------------------------------------------------------------- ------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again ------------------------------------------------------------------------------- ------- From plaurie <@t> benaroyaresearch.org Tue Sep 6 16:31:27 2005 From: plaurie <@t> benaroyaresearch.org (Patrick Laurie) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] (no subject) Message-ID: I am trying to stain for motor neurons on frozen 8um sections for LCM. I have been working with the .5% Cresyl Violet Acetate stain (Carson's version of Vacca's stain) and all of it's derivatives. I am trying to counter the differentiation of the alcohols used to completely dehydrate the sections and leaving very little stain left. I am aiming for RNA preservation, so length of time in the stain is important. I would appreciate any tips. Thanks, Patrick Laurie HT (ASCP) Neurogenomics Laboratory Benaroya Research Institute 1201 9th Ave Seattle, Wa 98101 (206) 341-0681 From cfavara <@t> niaid.nih.gov Tue Sep 6 16:55:46 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] recycling Message-ID: All, I have been asked to explore recycling my dehydrating and clearing agents. I currently use Propar[Anatech] and ETOH. I would be interested in any information and help. This is a research facility and we work mainly with prions and murine retroviruses. I am just beginning to think of the issues that my investigator may bring to the forefront so any insight would be helpful. Vendors welcome via e-mail only please! Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From Linresearch <@t> aol.com Tue Sep 6 18:50:32 2005 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] (no subject) Message-ID: <81.2f67d8b0.304f8548@aol.com> Hi, I am in need of a good Tunnel Kit that will work on FFPE rodent tissues. I was using the Roche kit with good results but the last 2 kits that I ordered have failed to stain anything. I would appreciate information on a kit that is working well and a protocol if possible. Thanks, Lin From josephnerk <@t> hotmail.com Tue Sep 6 19:53:58 2005 From: josephnerk <@t> hotmail.com (Joseph Nerk) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Re- histotech Message-ID: Hello i am a histotech with over 20 years experience, though i am trained in the Caribbean I have done a wide range of special stains, lots of frozen sections. Taught and trained lots of Histotechs. Did some training in Cancer Registry. trained on the Immunostainer as well as the Cerner lab program in Histopathology. My Histology final exam was two 3 hour essay type papers, an hour and a half paper in multiple choice of over 100 questions. Two 3 hour practicals one in which there was tissue and stains to be identified, serial sections, doing select special stains. Also did frozen sections, as well as fat stains. There was also an oral, where there was a team of pathologists asking quetion pertaining to the exam and related matters. _________________________________________________________________ Express yourself instantly with MSN Messenger! [1]MSN Messenger Download today it's FREE! References 1. http://g.msn.com/8HMAEN/2740??PS=47575 From Linresearch <@t> aol.com Wed Sep 7 05:17:46 2005 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Tunnel Kit Message-ID: <83.2f5ce427.3050184a@aol.com> Hi, I am in need of a good Tunnel Kit that will work on FFPE rodent tissues. I was using the Roche kit with good results but the last 2 kits that I ordered have failed to stain anything. I would appreciate information on a kit that is working Thanks, Lin From jstaruk <@t> masshistology.com Wed Sep 7 07:58:44 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Tunnel Kit In-Reply-To: <83.2f5ce427.3050184a@aol.com> Message-ID: <0IMG00NWH6PC2W2B@vms044.mailsrvcs.net> We use the Promega "DeadEnd Colorimetric TUNEL System" (#G7361) which works great on FFPE rodent tissue. Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linresearch@aol.com Sent: Wednesday, September 07, 2005 6:18 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Tunnel Kit Hi, I am in need of a good Tunnel Kit that will work on FFPE rodent tissues. I was using the Roche kit with good results but the last 2 kits that I ordered have failed to stain anything. I would appreciate information on a kit that is working Thanks, Lin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From venkino_vet <@t> rediffmail.com Wed Sep 7 08:17:55 2005 From: venkino_vet <@t> rediffmail.com (kamala venkatesh) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Diff quick Message-ID: <20050907131755.10133.qmail@webmail18.rediffmail.com> ? Dear Friends, This is Dr.Kamalavenkatesh from Wockhardt research center, Mumbai, India. We are in need of Diff- Quick Stain set. We are in need of the information regarding, the manufacturer and th esuppliers of the same from India. Thanks and regards Dr.Kamalavenkatesh From Angel.Enniss <@t> pfizer.com Wed Sep 7 08:34:58 2005 From: Angel.Enniss <@t> pfizer.com (Enniss, Angel) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] H&E Message-ID: I have been working on getting a new staining protocol or new Hematoxyilin. My supervisor does not like the blue background and she thinks we can do better than Harris. I am doing a demo on a linear stainer. I want to try lowering our times down to 30 seconds and using Harris with acid. 6 stations-xylene 2 stations-100% 1 station-95% 2 stations-running tap water 2 stations-Hematoxylin 3 stations-tap water 1 station-80% 1 station-Eosin 2 Station-95% 3 Station-100% 2 Stations-Xylene Does anyone else have any better ideas for an automated stainer or linear stainer. If you could send protocols that would be useful. Thanks so much. Angel C. Enniss, HT (ASCP) World Wide Safety Sciences AA e-mail: angel.enniss@pfizer.com Tel 734 622 3154 Fax 734 622 3866 LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From Kemlo.Rogerson <@t> elht.nhs.uk Wed Sep 7 08:44:36 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] H&E Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698F87@elht-exch1.xelht.nhs.uk> You don't have an acid/ alcohol step? Either use a fresh progressive haematoxylin step or introduce an acid/ alcohol step to remove the 'overstaining', then 'blue'. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Enniss, Angel Sent: 07 September 2005 14:35 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] H&E I have been working on getting a new staining protocol or new Hematoxyilin. My supervisor does not like the blue background and she thinks we can do better than Harris. I am doing a demo on a linear stainer. I want to try lowering our times down to 30 seconds and using Harris with acid. 6 stations-xylene 2 stations-100% 1 station-95% 2 stations-running tap water 2 stations-Hematoxylin 3 stations-tap water 1 station-80% 1 station-Eosin 2 Station-95% 3 Station-100% 2 Stations-Xylene Does anyone else have any better ideas for an automated stainer or linear stainer. If you could send protocols that would be useful. Thanks so much. Angel C. Enniss, HT (ASCP) World Wide Safety Sciences AA e-mail: angel.enniss@pfizer.com Tel 734 622 3154 Fax 734 622 3866 LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vanann702 <@t> skmc.gov.ae Wed Sep 7 08:44:35 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Diff quick Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CBBB@SKMCEMAIL.skmc.gov.ae> Do a 'google' search on the name of any chemical/stain/dye you need, follow the links and then look for local suppliers vendors etc. im in abu dhabi and we use the thermo Shandon stuff - works pretty good for us Good luck Annie in arabia -----Original Message----- From: kamala venkatesh [mailto:venkino_vet@rediffmail.com] Sent: Wednesday, September 07, 2005 5:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Diff quick ? Dear Friends, This is Dr.Kamalavenkatesh from Wockhardt research center, Mumbai, India. We are in need of Diff- Quick Stain set. We are in need of the information regarding, the manufacturer and th esuppliers of the same from India. Thanks and regards Dr.Kamalavenkatesh _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Evelyn.Flynn <@t> childrens.harvard.edu Wed Sep 7 08:41:37 2005 From: Evelyn.Flynn <@t> childrens.harvard.edu (Flynn, Evelyn) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] TUNEL Kit References: <83.2f5ce427.3050184a@aol.com> Message-ID: Dear Lin, For many years I have used the TUNEL kits which are distributed presently by Chemicon. Specimens have included FFPE rodent tissues. Good luck, Evelyn Flynn, M.A. Children's Hospital, Boston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Linresearch@aol.com Sent: Wed 9/7/2005 6:17 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Tunnel Kit Hi, I am in need of a good Tunnel Kit that will work on FFPE rodent tissues. I was using the Roche kit with good results but the last 2 kits that I ordered have failed to stain anything. I would appreciate information on a kit that is working Thanks, Lin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Debbiejsiena <@t> aol.com Wed Sep 7 09:14:37 2005 From: Debbiejsiena <@t> aol.com (Debbiejsiena@aol.com) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Help Wanted Lead Tech (Dallas, Texas) Message-ID: <12d.643b2f0d.30504fcd@aol.com> Good Morning All Tissue Techniques Path Labs in Dallas Texas is looking for a lead tech. We have a current opening that needs to be filled quickly, we are taking in more new business, this new business will leave us short handed until we can staff up. If you are interested please respond to any of the following: Sincerely; Debbie Siena HT ASCP (QIHC) Office: 972-241-6277, Fax: 972-241-6277, Cell: 817-228-8023, Cell: 520-360-3013 E-mail: Tissuetech@juno.com From ROrr <@t> enh.org Wed Sep 7 09:21:30 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH meet and Greet Message-ID: Hello everyone, Our Florida friends have recommended the RIVERWATCH LOUNGE at the MARINA MARRIOTT MONDAY,SEPTEMBER 12, from 6:30 to 8:00pm, for our casual meeting. This is the same hotel as the Thermo Party, and as I understand it, there is shuttle service between the hotels. You'll see a sign or perhaps a person with a computer on their head directing you to the area where we'll be congregating. Based on the high number of responses, there should be around 50 of us there! Thanks, Becky Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 From NSEARCY <@t> swmail.sw.org Wed Sep 7 09:32:18 2005 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] "Grossing" Tech Message-ID: Can anyone supply me with salary suggestions regarding what one pays for a "grossing tech?" This person is performing high complexity testing according to CLIA (associates degree, etc. + competency / training by PA or pathologist). I have a justification to present ASAP. Thanks From Angel.Enniss <@t> pfizer.com Wed Sep 7 09:39:49 2005 From: Angel.Enniss <@t> pfizer.com (Enniss, Angel) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] H&E Message-ID: I am trying to get procedures for linear stainers together. If anyone can give me a copy , fax or mail, would be great full!! Thanks much Angel C. Enniss, HT (ASCP) World Wide Safety Sciences AA e-mail: angel.enniss@pfizer.com Tel 734 622 3154 Fax 734 622 3866 LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From anne.lewin <@t> bms.com Wed Sep 7 10:47:27 2005 From: anne.lewin <@t> bms.com (Anne C Lewin) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: <431F0B8F.6030100@bms.com> Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... From tpmorken <@t> labvision.com Wed Sep 7 11:05:12 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D0D5@usca0082k08.labvision.apogent.com> Weather.com shows scattered T-storms, no flight delays (on Friday at least...). Ophelia is headed north, away from south florida. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C Lewin Sent: Wednesday, September 07, 2005 8:47 AM To: Histonet Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... From jqb7 <@t> cdc.gov Wed Sep 7 11:07:12 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: I thought it was way north...around Cape Canaveral and moving to the northwest........ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C Lewin Sent: Wednesday, September 07, 2005 11:47 AM To: Histonet Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... From jkiernan <@t> uwo.ca Wed Sep 7 11:09:34 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:33 2005 Subject: Cresyl violet. Was [Histonet] (no subject) References: Message-ID: <431F10BE.B82B6023@uwo.ca> You identify your dye as cresyl violet acetate. Does the bottle of dye powder carry a little label indicating that the batch was certified by the Biological Stain Commission (BSC)? If your dye is not from a certified batch it may be no good. The cresyl violet name has been attached to a variety of mixtures of cationic oxazine dyes. See Conn's Biological Stains for more information. Certified cresyl violet will always work as a Nissl stain if it's used in the technique with which it was tested in the BSC's lab. This has been published on various occasions, most recently by Penney et al (2002) in Biotechnic & Histochemistry 77:237-275. The second of the two cresyl violet methods in Freida Carson's textbook (which you cite in your email) is closely similar to the BSC's testing technique. Both procedures are explained in detail there (pp 159-161), with colour photos of the desired result. You are using cryostat sections rather than paraffin. The staining may become more controllable if you take the slides to 100% alcohol and back to water before staining. John Kiernan Anatomy, UWO London, Canada ____________________ Patrick Laurie wrote: > > I am trying to stain for motor neurons on frozen 8um sections for LCM. I > have been working with the .5% Cresyl Violet Acetate stain (Carson's version > of Vacca's stain) and all of it's derivatives. I am trying to counter the > differentiation of the alcohols used to completely dehydrate the sections > and leaving very little stain left. I am aiming for RNA preservation, so > length of time in the stain is important. I would appreciate any tips. > Thanks, > > Patrick Laurie HT (ASCP) > Neurogenomics Laboratory > Benaroya Research Institute > 1201 9th Ave > Seattle, Wa 98101 > (206) 341-0681 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Wed Sep 7 11:13:04 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: <20050907161304.8578.qmail@web30404.mail.mud.yahoo.com> Here is a cool link to the National Hurricane Center to watch for any problems with weather in Florida this weekend. I have been questioning the logic of this location for a September convention. http://www.nhc.noaa.gov/ Stephen From Diane.Gladney <@t> se.amedd.army.mil Wed Sep 7 11:15:41 2005 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: <4D55B2E997EFAE4DA6081DDE100B83027C1149@amedmlsermc133.amed.ds.army.mil> I am worried about my drive down from Columbia, SC on Friday. It looks like I may driving into a lot of rain and wind. Hopefully, this storm stays just a tropical storm and not a hurricane. I have a feeling that my 10-11 hour drive will turn into a very long trip.....15 hours or more. I am suppose to be in the Liaison Meeting at 6:00 pm on Friday, but I may miss it because of driving delays. Please keep me in your thoughts and prayers on Friday as I drive. I plan on leaving around 4 am. Thanks for indulging me and my concerns. Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Street P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C Lewin Sent: Wednesday, September 07, 2005 11:47 AM To: Histonet Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... From Janet.Bonner <@t> FLHOSP.ORG Wed Sep 7 11:10:21 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB43CF@fh2k093.fhmis.net> Well, it's this way..... the storm dumping rain on us now should have moved north ...unless it goes out to sea, loops around and comes back. Hurricane Nate has slowed but doesn't look like a problem. We usually get lots of notice (2-3 days) so if someone murmers evacuate, do so QUICKLY in a northerly direction..and then you'll be OK. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet Sent: 9/7/2005 11:47 AM Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... <> From Jackie.O'Connor <@t> abbott.com Wed Sep 7 11:18:11 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: Don't forget about the other two brewing in the Atlantic - Nate and Maria. "Bartlett, Jeanine" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/07/2005 11:07 AM To: "Anne C Lewin" , "Histonet" cc: Subject: RE: [Histonet] NSH and Hurricane Ophelia I thought it was way north...around Cape Canaveral and moving to the northwest........ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C Lewin Sent: Wednesday, September 07, 2005 11:47 AM To: Histonet Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jfish <@t> gladstone.ucsf.edu Thu Sep 8 11:24:46 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia In-Reply-To: <431F0B8F.6030100@bms.com> References: <431F0B8F.6030100@bms.com> Message-ID: Hi Anne and NSH symposiumers, I heard this morning that it is just a tropical depression bringing rain but not much else. Check out http://www.nhc.noaa.gov/ , the site has maps and lots of neat stuff about the current storms. It looks like the brunt of this storm will actually be north of Fort Lauderdale. Now, I am from California, we don't have "weather". I picture myself packing twice as many clothes as I'll need, just in case! Gotta remember an umbrella! See you all there, Jo Dee At 11:47 AM -0400 9/7/05, Anne C Lewin wrote: >Anyone heard any news about the weather in Ft Lauderdale coming up? >Flying in on Friday, hoping that there still will be a >conference...... > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-5766 Fax: (415) 355-0824 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From BHart <@t> asterand.com Wed Sep 7 11:46:07 2005 From: BHart <@t> asterand.com (Brian Hart) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Old Paraffin Blocks Message-ID: <1B0B60D56CA6A440A240E866BA3458D8055D6C@ATL1VEXC017.usdom004.tco.tc> If you are destroying your old paraffin blocks or paying to have it done for you - STOP! We would like to work with you. Asterand, Inc. has a program whereby we help salvage tissue blocks and associated clinical data no longer required to be maintained by CAP and JCAHO. If you would like to add revenue to your hospital rather than lose it, we would be interested in speaking with you. Please contact either Brian Hart - bhart@asterand.com or Neil Mucci - nmucci@asterand.com if you would like more information. Thank you, Brian Hart Asterand, Inc. This e-mail message, together with any attachments, contains information belonging to Asterand, Inc. that may be confidential, proprietary, copyrighted and/or legally protected or privileged, and is intended for the sole use of the individual or entity named on this message. If you are not the intended recipient, and have received this e-mail message in error, please immediately return this message to its original sender and permanently delete it from your electronic and paper records. Thank You. From jqb7 <@t> cdc.gov Wed Sep 7 12:14:30 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: Track them all here: http://www.hurricanetrack.com/ _____ From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: Wednesday, September 07, 2005 12:18 PM To: Bartlett, Jeanine Cc: Anne C Lewin; Histonet; histonet-bounces@lists.utsouthwestern.edu Subject: RE: [Histonet] NSH and Hurricane Ophelia Don't forget about the other two brewing in the Atlantic - Nate and Maria. "Bartlett, Jeanine" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/07/2005 11:07 AM To: "Anne C Lewin" , "Histonet" cc: Subject: RE: [Histonet] NSH and Hurricane Ophelia I thought it was way north...around Cape Canaveral and moving to the northwest........ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C Lewin Sent: Wednesday, September 07, 2005 11:47 AM To: Histonet Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Wed Sep 7 12:21:35 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: I wonder why all these destructive gushes of wind are named after wome...........ouch. Secretary just came in:-) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: 07 September 2005 18:15 To: Jackie M O'Connor Cc: Anne C Lewin; histonet-bounces@lists.utsouthwestern.edu; Histonet Subject: RE: [Histonet] NSH and Hurricane Ophelia Track them all here: http://www.hurricanetrack.com/ _____ From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: Wednesday, September 07, 2005 12:18 PM To: Bartlett, Jeanine Cc: Anne C Lewin; Histonet; histonet-bounces@lists.utsouthwestern.edu Subject: RE: [Histonet] NSH and Hurricane Ophelia Don't forget about the other two brewing in the Atlantic - Nate and Maria. "Bartlett, Jeanine" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/07/2005 11:07 AM To: "Anne C Lewin" , "Histonet" cc: Subject: RE: [Histonet] NSH and Hurricane Ophelia I thought it was way north...around Cape Canaveral and moving to the northwest........ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C Lewin Sent: Wednesday, September 07, 2005 11:47 AM To: Histonet Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Wed Sep 7 12:32:01 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia In-Reply-To: References: Message-ID: <6.1.1.1.2.20050907133035.019a9610@mail.vet.upenn.edu> Actually several years ago this was mentioned and they are now rotated one is female and the next is a male name through the alphabet. We should all share the blame for those outbursts. Pam At 01:21 PM 9/7/2005, Marshall Terry Dr, Consultant Histopathologist wrote: >I wonder why all these destructive gushes of wind are named after >wome...........ouch. Secretary just came in:-) > >Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path > Consultant Pathologist > Rotherham General Hospital > South Yorkshire > England > terry.marshall@rothgen.nhs.uk > >-----Original Message----- >From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] >Sent: 07 September 2005 18:15 >To: Jackie M O'Connor >Cc: Anne C Lewin; histonet-bounces@lists.utsouthwestern.edu; Histonet >Subject: RE: [Histonet] NSH and Hurricane Ophelia > > >Track them all here: > >http://www.hurricanetrack.com/ > > _____ > >From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] >Sent: Wednesday, September 07, 2005 12:18 PM >To: Bartlett, Jeanine >Cc: Anne C Lewin; Histonet; histonet-bounces@lists.utsouthwestern.edu >Subject: RE: [Histonet] NSH and Hurricane Ophelia > > > >Don't forget about the other two brewing in the Atlantic - Nate and >Maria. > > > > > > "Bartlett, Jeanine" >Sent by: histonet-bounces@lists.utsouthwestern.edu > >09/07/2005 11:07 AM > > > > To: "Anne C Lewin" , "Histonet" > > cc: > Subject: RE: [Histonet] NSH and Hurricane Ophelia > > > >I thought it was way north...around Cape Canaveral and moving to the >northwest........ > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C >Lewin >Sent: Wednesday, September 07, 2005 11:47 AM >To: Histonet >Subject: [Histonet] NSH and Hurricane Ophelia > >Anyone heard any news about the weather in Ft Lauderdale coming up? >Flying in on Friday, hoping that there still will be a conference...... > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From sluhisto <@t> yahoo.com Wed Sep 7 12:59:28 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Class A pipettes/cylinders Message-ID: <20050907175928.31036.qmail@web51015.mail.yahoo.com> Hello All: I know many of you are gearing up for NSH and I hope to meet some of you there, but I am in need of vendor information for class A pipettes/cylinders. If anyone has some info to share, I would be grateful. Thanks bunches. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From MICHAEL.OWEN <@t> fda.gov Wed Sep 7 13:04:48 2005 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: Recommended Hurricane News Web Sites National Hurricane Center / Tropical Prediction Center Hurricane and Storm Tracking for Atlantic and Pacific Oceans The Weather Channel CNN.com Fox News Network MSNBC News CDC: Emergency Preparedness and Response CDC: Emergency Preparedness and Response / Hurricanes Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From mucram11 <@t> comcast.net Wed Sep 7 13:10:57 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: <090720051810.4340.431F2D30000D9667000010F42207021553CECE030E9D0C9A03@comcast.net> Thanks, Michael. If we can't find at one of these sites we are in trouble. The NOAA site is great. Pam Marcum -------------- Original message -------------- > Recommended Hurricane News Web Sites > > National Hurricane Center / Tropical Prediction Center > > > Hurricane and Storm Tracking for Atlantic and Pacific Oceans > > > The Weather Channel > > > CNN.com > > > Fox News Network > > > MSNBC News > > > CDC: Emergency Preparedness and Response > > > CDC: Emergency Preparedness and Response / Hurricanes > > > > > > Michael P. Owen, Regulatory Microbiologist > U.S. FDA Pacific Regional Lab Northwest > 22201 23rd Drive SE Bothell, WA 98021-4421 > Phone: 425-483-4865 E-Mail: michael.owen@fda.gov > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ajohnson <@t> aipathology.com Wed Sep 7 13:10:23 2005 From: ajohnson <@t> aipathology.com (Amy Johnson) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Ventana Benchmark Message-ID: <016A64931E1ED511B12C0002B30260805233F4@SERV001> We were wondering what should be the pH of the EZ prep solution used on the Benchmark machine. We have been pH'ing it at 6 but have been told it should really be at 8. Please let us know hat others are using Thanks Amy Johnson From la.sebree <@t> hosp.wisc.edu Wed Sep 7 13:52:43 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Ventana Benchmark Message-ID: We were told not to pH the EZ Prep because its a detergent so we never have. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Johnson Sent: Wednesday, September 07, 2005 1:10 PM To: Histonet (E-mail) Subject: [Histonet] Ventana Benchmark We were wondering what should be the pH of the EZ prep solution used on the Benchmark machine. We have been pH'ing it at 6 but have been told it should really be at 8. Please let us know hat others are using Thanks Amy Johnson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Wed Sep 7 14:07:28 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB43D0@fh2k093.fhmis.net> Actually, not all of them are- we've had Andrew, Charley, Dennis, Hugo, Ivan....alot of the horrible one's have been guy names! This year I'm sure we'll get to the "T"s for you! @:) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Bartlett, Jeanine; Jackie M O'Connor Cc: Anne C Lewin; histonet-bounces@lists.utsouthwestern.edu; Histonet Sent: 9/7/2005 1:21 PM Subject: RE: [Histonet] NSH and Hurricane Ophelia I wonder why all these destructive gushes of wind are named after wome...........ouch. Secretary just came in:-) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: 07 September 2005 18:15 To: Jackie M O'Connor Cc: Anne C Lewin; histonet-bounces@lists.utsouthwestern.edu; Histonet Subject: RE: [Histonet] NSH and Hurricane Ophelia Track them all here: http://www.hurricanetrack.com/ _____ From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: Wednesday, September 07, 2005 12:18 PM To: Bartlett, Jeanine Cc: Anne C Lewin; Histonet; histonet-bounces@lists.utsouthwestern.edu Subject: RE: [Histonet] NSH and Hurricane Ophelia Don't forget about the other two brewing in the Atlantic - Nate and Maria. "Bartlett, Jeanine" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/07/2005 11:07 AM To: "Anne C Lewin" , "Histonet" cc: Subject: RE: [Histonet] NSH and Hurricane Ophelia I thought it was way north...around Cape Canaveral and moving to the northwest........ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C Lewin Sent: Wednesday, September 07, 2005 11:47 AM To: Histonet Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jennifer.l.hofecker <@t> Vanderbilt.Edu Wed Sep 7 17:01:36 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Ophelia and the s/c (slightly OT) Message-ID: <898D946569A27444B65667A49C074052075494@mailbe06.mc.vanderbilt.edu> This might be a good occasion to pack those wellies that were discussed last week! Maybe we could add a new category to the Tshirt contest for the best ones :) See y'all in FLA, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From liz <@t> premierlab.com Wed Sep 7 17:19:50 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] peptide getting into tumor cells Message-ID: <000001c5b3fa$43642680$a7d48a80@AMY> Hello everyone I have a unique question. I'm working with a client who is trying to duplicate some previously published work. They have a peptide that is supposed to target tumor cells (internalized by tumor cells). What they are looking at is targeted drug delivery. In publication they took unfixed frozen sections of human tumors and incubated them with a FITC labeled peptide for 12 hours at room temperature in a humidity chamber and then rinsed in PBS with glycine for 48 hours with multiple changes. The materials and methods in the publication were very minimal. The client followed their staining protocol but used both fixed and non fixed frozen sections. The staining pattern was diffuse. I made some initial suggestions of looking at fixed unstained slides and then a FITC labeled peptide that had been scrambled (not the same sequence) using the same method. My question is about the technique that is being used. I have done a lot of immunohistochemistry and immunofluoresence but the concept here is a bit different in that we are trying to determine if this peptide actually is internalized by tumor cells. To me the incubation time seems long (these are 4 micron sections) and they did not use a blocking reagent to avoid charge related binding. If anyone has done anything like this before and has any suggestions that would be great. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From chengkaz <@t> uci.edu Wed Sep 7 17:27:58 2005 From: chengkaz <@t> uci.edu (Chengkang ZHANG) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Re: GnRH Staining Message-ID: <431F696E.6000603@uci.edu> Dear All, I am doing IHC against GnRH on mouse tissue. Here are some pics of my staining, one pic shows staining at ME (median eminence, [1]http://www.villagephotos.com/members/viewimage.asp?id_=13978262) and one at LH (Lateral Hypothalamus, [2]http://img.villagephotos.com/p/2005-9/1074408/20050901-P7WTLA_resiz e.jpg). I am new to the field of GnRH and are not sure if the staining are real. In most literature, GnRH staining on ME have much less cell body staining that what I got here, and at LH their staining are more close to the 3V. I hope experts on this list can look at the images and tell me if the staining is real. Your help is greatly appreciated. CK -- ================================== Chengkang Zhang Ph.D. Room 357, MedSurge II Department of Pharmacology University of California, Irvine Irvine, CA 92697-4625 Email: [3]chengkaz@uci.edu Tel: (949)-824-1902 (lab) References 1. http://img.villagephotos.com/p/2005-9/1074408/20050901-P7WTME20x_resize.jpg 2. http://img.villagephotos.com/p/2005-9/1074408/20050901-P7WTLA_resize.jpg 3. mailto:chengkaz@uci.edu From gangli.phd <@t> gmail.com Wed Sep 7 18:01:14 2005 From: gangli.phd <@t> gmail.com (Gang Li) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Absorption Spectra of Stained Specimen Message-ID: <120b7bf05090716013c08632c@mail.gmail.com> Hello, Could anyone give me some pointers to the absorption spectra of tissue specimens stained with common dyes? Thanks a lot. Gang Li From ja.mitchell <@t> hosp.wisc.edu Wed Sep 7 20:31:04 2005 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Red Cross Pledge Message-ID: <583D3E9A1E843445BD54E461D1A2F6F31381AEF0@uwhis-xchng2.hosp.wisc.edu> On behalf of the Board of Directors for the Wisconsin Histology Society I would like to announce that WHS has pledged a $1000 donation to the Hurricane Katrina Disaster Relief Fund C/O the American Red Cross. If you have not done so already; we would like to encourage & invite other state or regional histology societies to make similar donations to assist the relief efforts in New Orleans, southern Mississippi and surrounding areas. Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Neuromuscular Laboratory Madison, WI 53792 From MGomez <@t> ameripath.com Wed Sep 7 22:47:21 2005 From: MGomez <@t> ameripath.com (Gomez, Milton) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Automated microtomes Message-ID: <98E6E88F806B194A809BA1D62B9FA2CD2E774E@TXCLMAIL01.ameripath.local> Dear Histonetters: I am interested in getting automated microtomes fo my lab. What is your experience and what do you recommend? I need to convince my manager and give him reasons for going automated. Thanks in advance, MG From Terry.Marshall <@t> rothgen.nhs.uk Thu Sep 8 06:35:21 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia Message-ID: There's nothing like displaying your ignorance in public:-) Thanks to all those that put me right. I am dismayed that I missed such gems as hurricane Claude and hurricane Jeremy. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] Sent: 07 September 2005 20:07 To: Marshall Terry Dr, Consultant Histopathologist; 'histonet-bounces@lists.utsouthwestern.edu '; 'Bartlett, Jeanine '; 'Jackie M O'Connor ' Cc: 'Anne C Lewin '; 'Histonet ' Subject: RE: [Histonet] NSH and Hurricane Ophelia Actually, not all of them are- we've had Andrew, Charley, Dennis, Hugo, Ivan....alot of the horrible one's have been guy names! This year I'm sure we'll get to the "T"s for you! @:) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Bartlett, Jeanine; Jackie M O'Connor Cc: Anne C Lewin; histonet-bounces@lists.utsouthwestern.edu; Histonet Sent: 9/7/2005 1:21 PM Subject: RE: [Histonet] NSH and Hurricane Ophelia I wonder why all these destructive gushes of wind are named after wome...........ouch. Secretary just came in:-) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: 07 September 2005 18:15 To: Jackie M O'Connor Cc: Anne C Lewin; histonet-bounces@lists.utsouthwestern.edu; Histonet Subject: RE: [Histonet] NSH and Hurricane Ophelia Track them all here: http://www.hurricanetrack.com/ _____ From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: Wednesday, September 07, 2005 12:18 PM To: Bartlett, Jeanine Cc: Anne C Lewin; Histonet; histonet-bounces@lists.utsouthwestern.edu Subject: RE: [Histonet] NSH and Hurricane Ophelia Don't forget about the other two brewing in the Atlantic - Nate and Maria. "Bartlett, Jeanine" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/07/2005 11:07 AM To: "Anne C Lewin" , "Histonet" cc: Subject: RE: [Histonet] NSH and Hurricane Ophelia I thought it was way north...around Cape Canaveral and moving to the northwest........ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C Lewin Sent: Wednesday, September 07, 2005 11:47 AM To: Histonet Subject: [Histonet] NSH and Hurricane Ophelia Anyone heard any news about the weather in Ft Lauderdale coming up? Flying in on Friday, hoping that there still will be a conference...... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JEllin <@t> yumaregional.org Thu Sep 8 07:15:45 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Grossing tech Message-ID: This brings up a good point, how are grossing tech being paid, I know in my facility they only give you an extra dollar an hour than the other techs within my department. it turns out to be about 5% more a year, but how do you justify a wage of 23-28 dollars an hour to managment, when there is no information to support your claim. If any one out there can provide me information on this I would really appreciate the help. Usually the wage of the histotech that I have seen between 16 to 23 dollar range with 18-19 being mid level, but then supervisors make about 22-29 dollars an hour. Any help would be greatly apperciated. Jesus Ellin From cfavara <@t> niaid.nih.gov Thu Sep 8 07:46:48 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Automated microtomes Message-ID: I have 2 Leicas. They have saved my arm. I cannot recommend them highly enough! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Gomez, Milton [mailto:MGomez@ameripath.com] Sent: Wednesday, September 07, 2005 8:47 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated microtomes Dear Histonetters: I am interested in getting automated microtomes fo my lab. What is your experience and what do you recommend? I need to convince my manager and give him reasons for going automated. Thanks in advance, MG _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Thu Sep 8 08:34:13 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Automated microtomes In-Reply-To: References: Message-ID: <6.1.1.1.2.20050908093225.01977760@mail.vet.upenn.edu> I have used the Shandon Finesse and now have a Leica 2255 and both are great. Pam Marcum U PA Vet School At 08:46 AM 9/8/2005, Favara, Cynthia (NIH/NIAID) wrote: >I have 2 Leicas. They have saved my arm. I cannot recommend them highly >enough! > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is confidential >and may contain sensitive information. It should not be used by anyone who >is not the original intended recipient. If you have received this e-mail in >error please inform the sender and delete it from your mailbox or any other >storage devices. National Institute of Allergy and Infectious Diseases shall >not accept liability for any statements made that are sender's own and not >expressly made on behalf of the NIAID by one of its representatives > >-----Original Message----- >From: Gomez, Milton [mailto:MGomez@ameripath.com] >Sent: Wednesday, September 07, 2005 8:47 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Automated microtomes > >Dear Histonetters: >I am interested in getting automated microtomes fo my lab. What is your >experience and what do you recommend? I need to convince my manager and >give him reasons for going automated. > >Thanks in advance, >MG >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Thu Sep 8 08:38:45 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] NSH and Hurricane Ophelia In-Reply-To: References: Message-ID: <6.1.1.1.2.20050908093700.01988b88@mail.vet.upenn.edu> Here in the colonies we just like to spread the blame equally for the devastation. No matter the name these are horrible storms with consequences those of us who are safe can not even imagine. Pam Marcum At 07:35 AM 9/8/2005, Marshall Terry Dr, Consultant Histopathologist wrote: >There's nothing like displaying your ignorance in public:-) >Thanks to all those that put me right. >I am dismayed that I missed such gems as hurricane Claude and hurricane >Jeremy. > >Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path > Consultant Pathologist > Rotherham General Hospital > South Yorkshire > England > terry.marshall@rothgen.nhs.uk > >-----Original Message----- >From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] >Sent: 07 September 2005 20:07 >To: Marshall Terry Dr, Consultant Histopathologist; >'histonet-bounces@lists.utsouthwestern.edu '; 'Bartlett, Jeanine '; >'Jackie M O'Connor ' >Cc: 'Anne C Lewin '; 'Histonet ' >Subject: RE: [Histonet] NSH and Hurricane Ophelia > > > Actually, not all of them are- we've had Andrew, Charley, Dennis, Hugo, >Ivan....alot of the horrible one's have been guy names! This year I'm sure >we'll get to the "T"s for you! @:) > > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >To: Bartlett, Jeanine; Jackie M O'Connor >Cc: Anne C Lewin; histonet-bounces@lists.utsouthwestern.edu; Histonet >Sent: 9/7/2005 1:21 PM >Subject: RE: [Histonet] NSH and Hurricane Ophelia > >I wonder why all these destructive gushes of wind are named after >wome...........ouch. Secretary just came in:-) > >Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path > Consultant Pathologist > Rotherham General Hospital > South Yorkshire > England > terry.marshall@rothgen.nhs.uk > >-----Original Message----- >From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] >Sent: 07 September 2005 18:15 >To: Jackie M O'Connor >Cc: Anne C Lewin; histonet-bounces@lists.utsouthwestern.edu; Histonet >Subject: RE: [Histonet] NSH and Hurricane Ophelia > > >Track them all here: > >http://www.hurricanetrack.com/ > > _____ > >From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] >Sent: Wednesday, September 07, 2005 12:18 PM >To: Bartlett, Jeanine >Cc: Anne C Lewin; Histonet; histonet-bounces@lists.utsouthwestern.edu >Subject: RE: [Histonet] NSH and Hurricane Ophelia > > > >Don't forget about the other two brewing in the Atlantic - Nate and >Maria. > > > > > > "Bartlett, Jeanine" >Sent by: histonet-bounces@lists.utsouthwestern.edu > >09/07/2005 11:07 AM > > > > To: "Anne C Lewin" , "Histonet" > > cc: > Subject: RE: [Histonet] NSH and Hurricane Ophelia > > > >I thought it was way north...around Cape Canaveral and moving to the >northwest........ > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne C >Lewin >Sent: Wednesday, September 07, 2005 11:47 AM >To: Histonet >Subject: [Histonet] NSH and Hurricane Ophelia > >Anyone heard any news about the weather in Ft Lauderdale coming up? >Flying in on Friday, hoping that there still will be a conference...... > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vd38 <@t> georgetown.edu Thu Sep 8 09:24:07 2005 From: vd38 <@t> georgetown.edu (Vernon Dailey) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Priamry Antibodies to CD117 and SCA-1 Message-ID: <43204987.8040300@georgetown.edu> Hi All, I want to run an IHC on FFPE mouse tissue and am looking for some feedback on primary antibodies. Does anyone have experience with anti SCA-1 or CD117 antibodies for IHC. If so, please recommend a vendor and clone. Thanks a bunch. -Vernon From asmith <@t> mail.barry.edu Thu Sep 8 10:26:36 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Hurricane Ophelia Message-ID: <5D2189E74151CC42BEC02906BA8996322B90F0@exchsrv01.barrynet.barry.edu> Fortunately, Hurricane Ophelia is already so far north that the expected continued northward movement will take it into the path of the Prevailing Westerlies (Counter-Trade Winds) which will take it out to sea. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From gu.lang <@t> gmx.at Thu Sep 8 11:40:35 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] histotech journal Message-ID: Hi, Can anyone reccommend a histotech-journal? Easily available in Europe (specially Austria), that deals with basic histotechnology up to modern, new techniques. Not too scientific and research-oriented, more practical advises. Thank you Gudrun Lang From BMolinari <@t> heart.thi.tmc.edu Thu Sep 8 12:29:21 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Automated microtomes Message-ID: Leica...I love mine too. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pamela Marcum Sent: Thursday, September 08, 2005 8:34 AM To: Favara, Cynthia (NIH/NIAID); 'Gomez, Milton'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Automated microtomes I have used the Shandon Finesse and now have a Leica 2255 and both are great. Pam Marcum U PA Vet School At 08:46 AM 9/8/2005, Favara, Cynthia (NIH/NIAID) wrote: >I have 2 Leicas. They have saved my arm. I cannot recommend them highly >enough! > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is confidential >and may contain sensitive information. It should not be used by anyone who >is not the original intended recipient. If you have received this e-mail in >error please inform the sender and delete it from your mailbox or any other >storage devices. National Institute of Allergy and Infectious Diseases shall >not accept liability for any statements made that are sender's own and not >expressly made on behalf of the NIAID by one of its representatives > >-----Original Message----- >From: Gomez, Milton [mailto:MGomez@ameripath.com] >Sent: Wednesday, September 07, 2005 8:47 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Automated microtomes > >Dear Histonetters: >I am interested in getting automated microtomes fo my lab. What is your >experience and what do you recommend? I need to convince my manager and >give him reasons for going automated. > >Thanks in advance, >MG >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NSEARCY <@t> swmail.sw.org Thu Sep 8 12:35:06 2005 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Artisan Message-ID: Can I hear from users, especially in Texas. Having some problems and need to hear from users. Thanks Nita From dellav <@t> musc.edu Thu Sep 8 13:03:24 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] histotech journal Message-ID: Journal of Histotechnology www.nsh.org >>> "Gudrun Lang" 09/08/05 12:40PM >>> Hi, Can anyone reccommend a histotech-journal? Easily available in Europe (specially Austria), that deals with basic histotechnology up to modern, new techniques. Not too scientific and research-oriented, more practical advises. Thank you Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Thu Sep 8 19:07:20 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] histotech journal Message-ID: I strongly recommend the Journal of Histotechnology and Histologic (when you can get it) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Friday, 9 September 2005 2:41 AM To: Histonetliste (Histonetliste) Subject: [Histonet] histotech journal Hi, Can anyone reccommend a histotech-journal? Easily available in Europe (specially Austria), that deals with basic histotechnology up to modern, new techniques. Not too scientific and research-oriented, more practical advises. Thank you Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From dbpiontek <@t> hotmail.com Thu Sep 8 20:37:58 2005 From: dbpiontek <@t> hotmail.com (Denise Piontek) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Recommend an Iron Stain Message-ID: I am looking for opinions on the best iron stain for detecting iron artifact in tissue? We need to see if an iron based implant leeched into the tissue. We have conflicting opinions on which stain would be best. Thanks for your help, Denise Bland-Piontek, HTL(ASCP)CTBS(AATB) TOXiKON _________________________________________________________________ [1]Ready for kickoff? Sign up for Fox Fantasy Football powered by MSN. FREE to play! References 1. http://g.msn.com/8HMAENUS/2746??PS=47575 From kmb1904 <@t> aol.com Thu Sep 8 20:47:44 2005 From: kmb1904 <@t> aol.com (kmb1904@aol.com) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] fine needle aspirates assisting Message-ID: <8C782F79B63FBB8-145C-1D0AF@MBLK-M31.sysops.aol.com> Maybe someone can give me some advice....at our hospital..when there is a fine needle procedure the histotechs are asked to come down with a tray and make the slides....spray them ...etc. Is this normal procedure?? Any help will be appreciated. Thank you!! From moogoolt <@t> sbcglobal.net Fri Sep 9 00:20:43 2005 From: moogoolt <@t> sbcglobal.net (Lance Thomas) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] looking for entry level hitology position in orange county CA Message-ID: <000a01c5b4fe$3a6cf840$33b2ef45@lance1mr2t65yd> Can anyone recommend a lab where I can get some histology experience to become certified? I have been working as a MOHS histotech for 7 years and would like to expand my knowledge and skills in general histology. I am excellent at making frozen sections and have experience with rapid immunostaining, using the Ventana NexES IHC system. I would appreciate any help and or advice. Lance Thomas From jorge.tornero <@t> gmail.com Fri Sep 9 01:35:50 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Using denaturalized alcohols working with Technovit 7100 Message-ID: <8c964a79050908233524bbb899@mail.gmail.com> Hello, do you have experience using denaturalized alcohol in the dehydration proccess and infiltration with this resin? I am thinking about using this alcohol due to its lesser cost but I don't know if its denaturalizing agentes (0,3% diethyl phtalate and 2 ppm Bitrex aka Denatonium Benzoate) may have influence over the proccess Thank you very much for all Jorge Tornero IEO-CADIZ Spain From IKirbis <@t> onko-i.si Fri Sep 9 01:40:33 2005 From: IKirbis <@t> onko-i.si (=?iso-8859-2?Q?Kirbi=B9_Srebotnik_Irena?=) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] fine needle aspirates assisting Message-ID: our cytopathologists performs FNAB 2 per day, normal team have cytopathologist, nurse (taking care for the patients, cleaning the site for procedure, giving cytopathologist syringe with the needle and so on)and technicians taking care for proper labelling of smears and the requisition form, writing down sample special properties and suspending leftovers in cell medium for ancillary dg. method. We cannot imagine that technicians wouldn't participate and help in FNAB office Irena Srebotnik Kirbi?, MSc Institute of Oncology Dept. of Cytopathology Zalo?ka 2 1000 Ljubljana Slovenia phone +386 1 522 3826 fax +38615879400 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Friday, September 09, 2005 3:48 AM To: histonet@pathology.swmed.edu Subject: [Histonet] fine needle aspirates assisting Maybe someone can give me some advice....at our hospital..when there is a fine needle procedure the histotechs are asked to come down with a tray and make the slides....spray them ...etc. Is this normal procedure?? Any help will be appreciated. Thank you!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Sep 9 02:21:54 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] fine needle aspirates assisting Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD53E@elht-exch1.xelht.nhs.uk> Depending on the type of FNAC we either leave it to them, if they are experienced, or assist. Our Ultrasound guided Endoscopic FNAC of such lesions as pancreas, gut, thorax, etc., requires the presence of a BMS (me). I take the sample from the tip of the endoscope, with the help of a colleague and prepare air-dried slides. My colleague stains a few from each passage and 'washes' the tip in saline (it is re-used if needed on the same Patient), I look at a Giemsa stained prep and tell the Endoscopist if he has diagnostic material; if not he tries again, if he has we stop. This reduces the risk of 'failure' but sometimes can be a bit intimidating. I used to assist at CT directed FNAC lesions of the lung and it was a bit 'hairy' asking the Radiologist to perform another due to 'inadequacy' when you are watching a pneumo developing! Plus the microscope was heavy! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kmb1904@aol.com Sent: 09 September 2005 02:48 To: histonet@pathology.swmed.edu Subject: [Histonet] fine needle aspirates assisting Maybe someone can give me some advice....at our hospital..when there is a fine needle procedure the histotechs are asked to come down with a tray and make the slides....spray them ...etc. Is this normal procedure?? Any help will be appreciated. Thank you!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Sep 9 05:16:07 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] fine needle aspirates assisting Message-ID: It is to us. We have a tool cart set up specifically for this. It is one of those red mechanic's cart that you buy at Sears, and we have it stocked with all the supplies needed for FNA's. While there are histotechs in the department, we go to radiology and set up the cart and get everything ready for the pathologist. The cart makes it easy to take the scope with you. The pathologist does all the interacting with the radiologist, but we are there to do any staining of smears and handling of paperwork. On the cart we have; H&E, Diff-Quick, and PAP mini set-ups to please the palates of any of our 11 pathologists that may be called upon to do the needle. Now if we could get radiology to stop calling us five minutes AFTER they need us, it could be a great system. After hours, when we are not here, the pathologist on-call takes the cart and does the FNA by him/herself(we are not a 24hr operation). Hope this helps. Feel free to give me a call. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kmb1904@aol.com Sent: Thursday, September 08, 2005 8:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] fine needle aspirates assisting Maybe someone can give me some advice....at our hospital..when there is a fine needle procedure the histotechs are asked to come down with a tray and make the slides....spray them ...etc. Is this normal procedure?? Any help will be appreciated. Thank you!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From japoteete <@t> saintfrancis.com Fri Sep 9 11:37:23 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] C4d Message-ID: Does anyone have information they would be willing to share about C4d on FFPE tissue? I would especially be interested your methods of antigen retrieval with times, dilution used, and antibody incubation time. Any other comments would be welcome. Thanks in advance. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa OK. japoteete@saintfrancis.com ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From sa.drew <@t> hosp.wisc.edu Fri Sep 9 11:48:21 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] C4d Message-ID: We are using the Biogenesis C4d at 1:600 after 3 min HIER w/with pH 8.0 EDTA buffer in BioCare's DeCloaking Chamber. On our Ventana we use 32 min antibody titration with an amplification step. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Poteete, Jacquie A. Sent: Friday, September 09, 2005 11:37 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] C4d Does anyone have information they would be willing to share about C4d on FFPE tissue? I would especially be interested your methods of antigen retrieval with times, dilution used, and antibody incubation time. Any other comments would be welcome. Thanks in advance. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa OK. japoteete@saintfrancis.com ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From krat18 <@t> aol.com Fri Sep 9 12:13:36 2005 From: krat18 <@t> aol.com (krat18@aol.com) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] fine needle aspirates assisting In-Reply-To: References: Message-ID: <8C78378F32FBA42-640-8129@mblk-r30.sysops.aol.com> This is the way we do it here too. The pathologist charges according to how many slides he/she needs to read to determine adequacy---that's in addition to the passes, etc., that the radiologist makes. Karen Raterman krat18@aol.com SSMHC, St. Louis, MO -----Original Message----- From: GUTIERREZ, JUAN To: kmb1904@aol.com; histonet@pathology.swmed.edu Sent: Fri, 9 Sep 2005 05:16:07 -0500 Subject: RE: [Histonet] fine needle aspirates assisting It is to us. We have a tool cart set up specifically for this. It is one of those red mechanic's cart that you buy at Sears, and we have it stocked with all the supplies needed for FNA's. While there are histotechs in the department, we go to radiology and set up the cart and get everything ready for the pathologist. The cart makes it easy to take the scope with you. The pathologist does all the interacting with the radiologist, but we are there to do any staining of smears and handling of paperwork. On the cart we have; H&E, Diff-Quick, and PAP mini set-ups to please the palates of any of our 11 pathologists that may be called upon to do the needle. Now if we could get radiology to stop calling us five minutes AFTER they need us, it could be a great system. After hours, when we are not here, the pathologist on-call takes the cart and does the FNA by him/herself(we are not a 24hr operation). Hope this helps. Feel free to give me a call. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kmb1904@aol.com Sent: Thursday, September 08, 2005 8:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] fine needle aspirates assisting Maybe someone can give me some advice....at our hospital..when there is a fine needle procedure the histotechs are asked to come down with a tray and make the slides....spray them ...etc. Is this normal procedure?? Any help will be appreciated. Thank you!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Fri Sep 9 13:25:56 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] frozen section trichrome Message-ID: <20050909182556.18218.qmail@web90204.mail.scd.yahoo.com> I'm looking for a consistant protocol for a trichrome stain for FS. I tried 2 this morning on liver. Both were sectioned at 7u , air dried for 30 minutes, fixed in bouins for 30 minutes and stained. Extreamly red with a granular appearence. Only green was in the folds. My paraffin section control was fine. Steve --------------------------------- Click here to donate to the Hurricane Katrina relief effort. From histology <@t> gradymem.org Fri Sep 9 13:48:48 2005 From: histology <@t> gradymem.org (histology@gradymem.org) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] fine needle aspirates assisting Message-ID: We used to go to the radiology department to receive specimens. However, we are a small hospital pathology department with only 2 techs (day shift) and someone was not always available to assist. So we wrote up complete, simple instructions on how to handle the specimens, order them, and deliver them to the lab. They do a fine job without us. Angie Barnett, HTL(ASCP) Grady Memorial Hospital Pathology Department 2300 Iowa Street Chickasha, OK 73018 405/224-2258 ----- Original Message ----- From: kmb1904@aol.com Date: Thursday, September 8, 2005 8:47 pm Subject: [Histonet] fine needle aspirates assisting > Maybe someone can give me some advice....at our hospital..when > there is a fine needle procedure the histotechs are asked to come > down with a tray and make the slides....spray them ...etc. Is > this normal procedure?? Any help will be appreciated. Thank you!! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From RBARNHART <@t> summithealth.org Fri Sep 9 14:17:27 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] fine needle aspirates assisting Message-ID: We are also a small hospital (1 pathologist, 1.5 histo tech and 1 cyto tech). We are just the opposite, we did not use to go to but know we do. The pathologist goes and either myself or the cyto tech goes along to help. The pathologist will look at the slides and tell the radiologist if the sample was good enough for a diagnosis. This eliminates the patient having to be stuck several times if the first sample is diagnostic. Becky Barnhart, HT (ASCP) >>> 9/9/2005 2:48:48 PM >>> We used to go to the radiology department to receive specimens. However, we are a small hospital pathology department with only 2 techs (day shift) and someone was not always available to assist. So we wrote up complete, simple instructions on how to handle the specimens, order them, and deliver them to the lab. They do a fine job without us. Angie Barnett, HTL(ASCP) Grady Memorial Hospital Pathology Department 2300 Iowa Street Chickasha, OK 73018 405/224-2258 ----- Original Message ----- From: kmb1904@aol.com Date: Thursday, September 8, 2005 8:47 pm Subject: [Histonet] fine needle aspirates assisting > Maybe someone can give me some advice....at our hospital..when > there is a fine needle procedure the histotechs are asked to come > down with a tray and make the slides....spray them ...etc. Is > this normal procedure?? Any help will be appreciated. Thank you!! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From greg_optimum <@t> yahoo.com Fri Sep 9 15:00:45 2005 From: greg_optimum <@t> yahoo.com (Greg Lyness) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] distinguishing mouse vs. human cells Message-ID: <20050909200045.89785.qmail@web33609.mail.mud.yahoo.com> Dear histonetters, We are working with xenograft models of human pancreatic tumors implanted into the pancreas of nude mice. In order to prove that the tissue which is growing is actual human tumor (and not mouse cells) we want to stain for a specific antigen/marker to distinguish between human and mouse cells. Has anyone ever immunostained for any such antigen/marker, and if so, what did you use and from where did you obtain it? Thanks in advance, Greg Lyness Research Scientist Optimum Therapeutics Columbus, OH (614) 688-5885 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From weneng2004 <@t> yahoo.com Fri Sep 9 16:01:56 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] tumor histology Message-ID: <20050909210156.68661.qmail@web53413.mail.yahoo.com> Dear histonetter, I am trying to learn basic cell type of pancreatic tumor. I would really appreciate if anyone can provide a webside that has picture and text of it. I am new in this. Do all kinds of tumor have same type of structure? I will look up some books. But I need to have basic idea quickly now. Thanks, Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From PMonfils <@t> Lifespan.org Fri Sep 9 16:24:10 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] tumor histology Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175CF@lsexch.lsmaster.lifespan.org> There are several different types of pancreatic malignancies because the structure of the pancreas includes several distinct types of cells, any of which could give rise to a tumor. The following site has a lot of information on pancreatic cancers, including some views of histological sections: http://www.path.jhu.edu/pancreas/ From jseaton <@t> wlgore.com Fri Sep 9 20:02:38 2005 From: jseaton <@t> wlgore.com (Janella Seaton) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Janella Seaton/WLGORE is out of the office. Message-ID: I will be out of the office starting 09/09/2005 and will not return until 09/16/2005. I will respond to your message when I return. From tahseen <@t> brain.net.pk Fri Sep 9 13:10:24 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Nuclear staining Her2 Neu. Message-ID: <004701c5b569$c09c7600$972bfea9@m7c0y4> Dear All We have a Her2 Neu with irregular nuclear staining.It seems to be happening on breast needle cores. Could it be over or under fixation? If anyone has any information I would greatly appreciate it. Thanks in advance Regards Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. From Thomas.Crowell <@t> biogenidec.com Sat Sep 10 15:02:36 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Thomas Crowell/Cambridge/Biogen is out of the office. Message-ID: I will be out of the office starting 09/09/2005 and will not return until 09/15/2005. I will respond to your message when I return. From dbpiontek <@t> hotmail.com Sun Sep 11 11:38:08 2005 From: dbpiontek <@t> hotmail.com (Denise Piontek) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Processing Animal Tissues Message-ID: Does any one process multiple species on the same processing program? Interested to hear what species are combined? Thanks for your help, Denise Bland-Piontek, HTL(ASCP)CTBNS(AATB) Histology Supervisor TOXiKON _________________________________________________________________ [1]Find e-mail and documents on your PC instantly with the new MSN Search ToolbarFREE! References 1. http://g.msn.com/8HMAENUS/2737??PS=47575 From dbpiontek <@t> hotmail.com Sun Sep 11 11:40:18 2005 From: dbpiontek <@t> hotmail.com (Denise Piontek) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Processing Eyes Message-ID: I know I have seen an eye expert on the histonet a few times. I am looking for information on eye fixation and processing for multiple species. Opinions seem to be quite varied. Thanks for your help, Denise Bland-Piontek, HTL(ASCP)CTBNS(AATB) Histology Supervisor TOXiKON _________________________________________________________________ [1]Make FREE PC-to-PC calls with MSN Messenger. Get it now! References 1. http://g.msn.com/8HMAENUS/2749??PS=47575 From Jason.PALMER <@t> svhm.org.au Sun Sep 11 20:54:52 2005 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] RE: distinguishing between mouse and human cells Message-ID: We have used an anti human mitochondrial Ab to distinguish between human and mouse cells which worked quite nicely. Ours came from Calbiochem, although i am fairly sure they are no longer manufacturing it. There may be other companies producing a similar product though. Jason Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 fax +61 3 9416 0926 email: palmerj@svhm.org.au Message: 6 Date: Fri, 9 Sep 2005 13:00:45 -0700 (PDT) From: Greg Lyness Subject: [Histonet] distinguishing mouse vs. human cells To: histonet@lists.utsouthwestern.edu Message-ID: <20050909200045.89785.qmail@web33609.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Dear histonetters, We are working with xenograft models of human pancreatic tumors implanted into the pancreas of nude mice. In order to prove that the tissue which is growing is actual human tumor (and not mouse cells) we want to stain for a specific antigen/marker to distinguish between human and mouse cells. Has anyone ever immunostained for any such antigen/marker, and if so, what did you use and from where did you obtain it? Thanks in advance, Greg Lyness Research Scientist Optimum Therapeutics Columbus, OH (614) 688-5885 __ Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From chitnis.c <@t> neu.edu Sun Sep 11 22:09:56 2005 From: chitnis.c <@t> neu.edu (Chinmay Chitnis) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] anti-bovine procollagen antibody Message-ID: <8369345.1126494596507.JavaMail.chitnis.c@neu.edu> hello, I am a researcher in Northeastern university at Boston. Can anybody suggest a source for anti-bovine procollagen N-terminus antibody. Chemicon International has the C-terminus antibody for bovine procollagen but it does not have N-terminus antibody for the same. Can any one help me out? Thanks Chinmay From eugenio.pittioni <@t> uniud.it Mon Sep 12 01:51:50 2005 From: eugenio.pittioni <@t> uniud.it (Eugenio PITTIONI) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Processing Animal Tissues In-Reply-To: References: Message-ID: <200509120851.51358.eugenio.pittioni@uniud.it> Alle 18:38, domenica 11 settembre 2005, Denise Piontek ha scritto: > Does any one process multiple species on the same processing program? > Interested to hear what species are combined? Thanks for your help, > Here at the Animal Science Department we usually process together sample from cats, dogs (60%), fish (of many species) (35%) and other (bovine, other pets) 5%) without problem. We use a particular processing program only for fish's larvae (very rapid: only 3 hours). Hope this can help Eugenio PITTIONI DVM Animal Science Department University o UDINE Friuli (ITALY) > Denise Bland-Piontek, HTL(ASCP)CTBNS(AATB) > Histology Supervisor > TOXiKON > _________________________________________________________________ > > [1]Find e-mail and documents on your PC instantly with the new MSN > Search ToolbarFREE! > > References > > 1. http://g.msn.com/8HMAENUS/2737??PS=47575 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From christelle.gerard <@t> novartis.com Mon Sep 12 03:31:02 2005 From: christelle.gerard <@t> novartis.com (christelle.gerard@novartis.com) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] anti integrin alpha v beta 3 on mouse tissue Message-ID: Hi all, I want to run IHC against integrin alpha v beta 3 on FFPE or FS mouse tissue to detect endothelial cells in tumor. Does anyone have experience witth this antigen? If yes, so please recommend a clone, a vendor and a protocol. Thanks in advance. Christelle From rjr6 <@t> psu.edu Mon Sep 12 08:16:43 2005 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Processing Animal Tissues Message-ID: I process a variety of species every day. Mainly bovine but also do many other farm species and lab animals (mice, rats, rabbits). I also process many avian species in the same run mostly chicken but have done everything from a sparrow to ostrich. I've not had any trouble with this and I've been here 18 years. Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Denise Piontek Sent: Sunday, September 11, 2005 12:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Processing Animal Tissues Does any one process multiple species on the same processing program? Interested to hear what species are combined? Thanks for your help, Denise Bland-Piontek, HTL(ASCP)CTBNS(AATB) Histology Supervisor TOXiKON _________________________________________________________________ [1]Find e-mail and documents on your PC instantly with the new MSN Search ToolbarFREE! References 1. http://g.msn.com/8HMAENUS/2737??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From portera203 <@t> yahoo.com Mon Sep 12 08:19:26 2005 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Processing Animal Tissues In-Reply-To: Message-ID: <20050912131926.42387.qmail@web34110.mail.mud.yahoo.com> Our laboratory processes most everything together without any difficulties. The only items we process separately are very large specimens which need extended processing times (half of a canine kidney, thats how large). Our routine is mouse, rat, feline, canine, porcine, bovine, equine....etc. Hope this helps out. Denise Piontek wrote: Does any one process multiple species on the same processing program? Interested to hear what species are combined? Thanks for your help, Denise Bland-Piontek, HTL(ASCP)CTBNS(AATB) Histology Supervisor TOXiKON _________________________________________________________________ [1]Find e-mail and documents on your PC instantly with the new MSN Search ToolbarFREE! References 1. http://g.msn.com/8HMAENUS/2737??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Amy S.Porter, HT(ASCP), QIHC Laboratory Supervisor Michigan State University - Department of Physiology Division of Human Pathology Investigative Histopathology Laboratory 2201 Biomedical Physical Sciences Room 2133 East Lansing, MI 48824-3320 Ph: 517-355-6475 ext 1480/1481 Fax: 517-432-1368 portera@msu.edu / www.humanpathology.msu.edu --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From MICHAEL.OWEN <@t> fda.gov Mon Sep 12 10:26:47 2005 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Off-Topic: Seattle Times Preparedness Article Message-ID: Could you survive a disaster here? By Jolayne Houtz and Emily Heffter Seattle Times consumer-affairs Reporters Sunday, September 11, 2005 - Page updated at 01:43 AM Printable Disaster Tip Sheet Seattle Times Sunday, September 11, 2005 Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From JCLARK <@t> kumc.edu Mon Sep 12 10:48:28 2005 From: JCLARK <@t> kumc.edu (Julie Clark) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Tim Plummer are you out there? Message-ID: Can you contact me please if you are? Thx, Julie Clark Supervisor, Molecular Pathology Laboratory Department of Pathology Room 2027 WHW Mailstop 3045 Univ. of Kansas Med. Center Kansas City, KS 66160 Phone: 913-588-7254 Page: 913-917-6052 FAX: 913-588-7073 From wragga <@t> nhlbi.nih.gov Mon Sep 12 11:04:19 2005 From: wragga <@t> nhlbi.nih.gov (Wragg, Andrew (NIH/NHLBI)) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] B-Galactosidase Immuno Fluorescence Message-ID: <729DD7AACF248D41AF691EA868BB734D0765E1@NIHCESMLBX6.nih.gov> I am trying to stain mouse femoral artery OCT frozen sections with anti B Galactosiadase antibodies for immuno fuorescent imaging. I am using the ROSA26 mouse which expresses B Gal as my positive control. I have not managed to find an antibody or set of conditions that work. I have tried: ab6161, ABCAM, Rabbit polyclonal MAB3468, Chemicon, Mouse monoclonal I have tried paraformaldehyde 4%, ice cold methanol and methanol/acetone mix. I have no problem with X Gal staining the tissues. Many thnaks for any help in advance Andrew Wragg Cardiovascular Branch NHLBI-NIH 10 Center Drive, MSC 8016 Building 10-CRC, 5-3232 5th floor east wing Bethesda, MD 20892 Tel: (+1) 301-451-4506 Fax: (+1) 301-451-7090 Email: wragga@nhlbi.nih.gov From cbass <@t> bidmc.harvard.edu Mon Sep 12 12:41:59 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] injection marker for the spinal cord Message-ID: <84DF217E-23B4-11DA-93E3-0003930771EE@bidmc.harvard.edu> Hello, I am working on an experiment where I will inject the spinal cord of rats. Is there a way to mark the injection site with a dye? Basically I want to do my injection, wait several weeks and collect the tissue. So the dye would have to last a while. Any suggestions would be appreciated. I am particularly interested in the best way to process the tissues for the recommended dye. Thanks, Caroline Bass From NSEARCY <@t> swmail.sw.org Mon Sep 12 14:13:16 2005 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Flow Cytometry Tech Message-ID: Anyone know of a Flow Tech looking for a position? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Division Manager, Anatomic Pathology 2401 S. 31st. Street 254-724-2438 Temple, Texas,76502 254-724-2438 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Anatomic Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From weneng2004 <@t> yahoo.com Mon Sep 12 18:21:39 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] calcium staining Message-ID: <20050912232139.37534.qmail@web53405.mail.yahoo.com> Dear histonetters, I was asked to stain calcium on FFPE arterial sections. Does anyone know what staining is that? Thanks in advance! Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From MGomez <@t> ameripath.com Mon Sep 12 20:18:26 2005 From: MGomez <@t> ameripath.com (Gomez, Milton) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Automated microtomes Message-ID: <98E6E88F806B194A809BA1D62B9FA2CD2E7757@TXCLMAIL01.ameripath.local> Dear Histonetters, What are your thoughts on injuries related to the use of automated microtomes? I have heard that the use of automated microtomes increases the risk of injury and in most cases these are more severe. May you share your experience? I do know of one case where the tech lost part of his finger (luckily he was able to have it re-attached and the finger was saved). How many labs out there have used automated microtomes and have reverted back to manual microtomes for safety reasons? Does safety take priority over ergonomic reasons? Thanks in advance for your reply, Milton From BMolinari <@t> heart.thi.tmc.edu Tue Sep 13 05:34:22 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Automated microtomes Message-ID: Milton, I also know of a finger amputation but the tech was in a hurry and as a result was careless. It was an operators error not a mechanical one. I also know of a tech that developed a problem from repeatedly pushing on the advance button. I have no experience with this type of automated microtome so I cannot comment on that problem; I use the foot pedal model. I use have both the automated and manual and prefer the automated, but it is personal decision. I think the benefits out weigh the risk. Hope this helps. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gomez, Milton Sent: Monday, September 12, 2005 8:18 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated microtomes Dear Histonetters, What are your thoughts on injuries related to the use of automated microtomes? I have heard that the use of automated microtomes increases the risk of injury and in most cases these are more severe. May you share your experience? I do know of one case where the tech lost part of his finger (luckily he was able to have it re-attached and the finger was saved). How many labs out there have used automated microtomes and have reverted back to manual microtomes for safety reasons? Does safety take priority over ergonomic reasons? Thanks in advance for your reply, Milton _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Tue Sep 13 06:52:29 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] immunofluorescence Message-ID: Hi, Does anyone have experience with immunofluorescence protocols in dog kidney? Thanks, Betsy Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) From dsnider <@t> shrinenet.org Tue Sep 13 06:58:20 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Arundo (Bamboo cane reeds) Message-ID: <84BE46B37B314D409C5A17B7BAB022D61D4CAC@IDC-EX-VS01.shriners.cc> Hello netters! This is an odd one.... But I need whatever input you may have. I have a co worker, who is a bassoonist,( also has a biology degree) He and a group of musicians have been trying to figure out a way to increase the life and use of the reeds used on the Bassoon. They are handmade and quite pricey, and last only about a week. They have been trying to embed these reeds in LR White resin + etoh. He says they have had little success. He also asked about something called DMSO. Does anyone know what he is talking about? Are there any histologists here who deal with plants? Wouldn't he need to infiltrate this material? The reeds are approx .05- 1.3 mm. Whatever is used cannot add thickness to the reed and must be safe for them to put in their mouth when they play. ( I am lost!) Any suggestions will be greatly appreciated......I told him I would post this and see what people may know... Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From vanann702 <@t> skmc.gov.ae Tue Sep 13 07:16:42 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Arundo (Bamboo cane reeds) Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D621@SKMCEMAIL.skmc.gov.ae> Personally, I would not put anything with DMSO plus resin in MY mouth - that's for sure Will not use the stuff in my lab - all wax has to be DMSO free!! Do a web search - it's a 'carrier' and helps the wax to penetrate tissue and can also assist the wax/polymers to be absorbed into the skin. Unless of course you wear gloves - but not all of us wear them 100%of the time when handling wax. - so the way I understand, it will 'assist the penetration' of what ever needs to penetrate - which is okay as long as that substance is desirable, like an emollient or a moisturizer!!!! ;-) Annieinarabia -----Original Message----- From: Snider, Deanna [mailto:dsnider@shrinenet.org] Sent: Tuesday, September 13, 2005 3:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Arundo (Bamboo cane reeds) Hello netters! This is an odd one.... But I need whatever input you may have. I have a co worker, who is a bassoonist,( also has a biology degree) He and a group of musicians have been trying to figure out a way to increase the life and use of the reeds used on the Bassoon. They are handmade and quite pricey, and last only about a week. They have been trying to embed these reeds in LR White resin + etoh. He says they have had little success. He also asked about something called DMSO. Does anyone know what he is talking about? Are there any histologists here who deal with plants? Wouldn't he need to infiltrate this material? The reeds are approx .05- 1.3 mm. Whatever is used cannot add thickness to the reed and must be safe for them to put in their mouth when they play. ( I am lost!) Any suggestions will be greatly appreciated......I told him I would post this and see what people may know... Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Tue Sep 13 08:37:46 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] endogenous peroxidases in paraffin In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE017175CF@lsexch.lsmaster.lifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE017175CF@lsexch.lsmaster.lifespan.org> Message-ID: Does anyone have any thoughts on how much endogenous peroxidase remains in tissue after paraffin processing, I have experienced certainly less than frozen sections for sure, but still I want some stories from personal experience. I am trying to decide whether I should try to exclude some glucose oxidase block on paraffin sections as it seems to be unnecessary in one of my protocols. Thanks, Andrea -- From deb.vaneyck <@t> phci.org Tue Sep 13 08:50:15 2005 From: deb.vaneyck <@t> phci.org (Van Eyck, Deb) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Fascin Message-ID: Hi all---one immuno question----is anyone using an IVD Fascin stain on their Hodgkins cases. My docs like this---but most of my sources are RUO. Any input is appreciated. Thanks- Deb VanEyck This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. From jfish <@t> gladstone.ucsf.edu Tue Sep 13 09:18:00 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Arundo (Bamboo cane reeds) In-Reply-To: <84BE46B37B314D409C5A17B7BAB022D61D4CAC@IDC-EX-VS01.shriners.cc> References: <84BE46B37B314D409C5A17B7BAB022D61D4CAC@IDC-EX-VS01.shriners.cc> Message-ID: Dear Deanna, Wow, this is interesting! I am a clarinet player and I guess I know a little about reeds, although clarinet reeds are a bit different in shape from bassoon reeds, but they are still made of bamboo. One of the reasons that bamboo is used is that they are flexible after soaking (the player walks around for about 15 minutes with the reed in their mouth to get the right amount of flexibility) but still stiff enough to vibrate when air is passed by them. Every player soaks to their own desired moisture depending on their ombusure (spelling probably wrong!), and usually during long rest periods during a concert they will resoak them or lick them often to keep them moist. All of this being said, I can't imagine anyone wanting to embed their reeds in a known carcinogenic (resin) or an alcohol (drying properties) or DMSO (will allow whatever was on the reed last to enter the bloodstream of the player when placed in the mouth). Besides, embedding them in a resin will not allow the soaking needed to make the reed flexible. The reeds are going to break down no matter how well they are treated. After all, they are plant matter and plant cells break down. He and his musician friends might want to look into making their own reeds (I had seen websites in the past that explained how to "do-it-yourself") and possibly researching new materials to make the reeds from. Maybe they will discover something that will benefit all musicians without giving them cancer of the tongue or cheek! Tell your friend good luck, Jo Dee On Tue, 13 Sep 2005 07:58:20 -0400 "Snider, Deanna" wrote: > Hello netters! This is an odd one.... But I need >whatever input you may > have. > I have a co worker, who is a bassoonist,( also has a >biology degree) He and > a group of musicians have been trying to figure out a >way to increase the > life and use of the reeds used on the Bassoon. They are >handmade and quite > pricey, and last only about a week. > > They have been trying to embed these reeds in LR White >resin + etoh. He says > they have had little success. He also asked about >something called DMSO. > Does anyone know what he is talking about? Are there >any histologists here > who deal with plants? Wouldn't he need to infiltrate >this material? The > reeds are approx .05- 1.3 mm. Whatever is used cannot >add thickness to the > reed and must be safe for them to put in their mouth >when they play. ( I am > lost!) > > Any suggestions will be greatly appreciated......I told >him I would post > this and see what people may know... > Shriners Hospital for Children > Research Dept. > Cincinnatti, Oh > 513-872-6388 > > CONFIDENTIALITY NOTICE: This e-mail communication and >any attachments may > contain confidential and privileged information for the >use of the > designated recipients. If you are not the intended >recipient, (or > authorized to receive for the recipient) you are hereby >notified that you > have received this communication in error and that any >review, disclosure, > dissemination, distribution or copying of it or its >contents is prohibited. > If you have received this communication in error, please >destroy all copies > of this communication and any attachments and contact >the sender by reply > e-mail or telephone (813) 281-0300. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ljb <@t> medicine.wisc.edu Tue Sep 13 09:12:07 2005 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:25:33 2005 Subject: Fwd: Re: [Histonet] endogenous peroxidases in paraffin Message-ID: >>> LaCinda Burchell 09/13/05 9:08 AM >>> Hello, and Good Morning; I've never seen differences in frozen versus paraffin tissues. Endogenous peroxidase is an important issue in bloody tissues (liver, spleen). If you are working with extremely fragile specimens low in red blood cells, and you've done a side-by-side demonstration of blocked and un-blocked specimens you should be ok. good luck! Cindy >>> "Andrea T. Hooper" 09/13/05 8:37 AM >>> Does anyone have any thoughts on how much endogenous peroxidase remains in tissue after paraffin processing, I have experienced certainly less than frozen sections for sure, but still I want some stories from personal experience. I am trying to decide whether I should try to exclude some glucose oxidase block on paraffin sections as it seems to be unnecessary in one of my protocols. Thanks, Andrea -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 From RSRICHMOND <@t> aol.com Tue Sep 13 11:11:20 2005 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Re: frozen section trichrome Message-ID: <75.4d4abc1b.30585428@aol.com> Steven Coakley (where?) is >>looking for a consistent protocol for a trichrome stain for [frozen sections].<< I'd recommend the one-step trichrome that's used for skeletal muscle frozen sections. Like practically every other stain, it's associated with the hallowed name of George Gomori. The muscle stain was developed by W. King Engel (neurologist) and Guy Cunningham (histotechnologist) at the NIH about forty years ago, and it's sometimes referred to as the Engel-Cunningham variant of the Gomori trichrome stain. I think it's commercially available. - The one-step trichrome modifications for stool parasitology probably aren't suitable, but you could try. Bob Richmond Gaston Memorial Hospital, Gastonia NC From JGordon <@t> cellmarque.com Tue Sep 13 11:12:05 2005 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Fascin Message-ID: Deb, there are many IHC companies that offer Fascin, as it is one of the more popular Hodgkin's Lymphoma markers. I am not sure which companies have RUO or IVD status on their Fascin. Cell Marque's is compatible with any automation, and it is an IVD antibody. http://www.cellmarque.com/products/productdetail.php?id=82&alpha= If there is anything we can do to assist you, please let us know. Jeff Gordon Cell Marque Corp. 1-800-665-7284 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Van Eyck, Deb Sent: Tue 9/13/2005 8:50 AM To: 'histonet@lists.utsouthwestern.edu' Cc: Subject: [Histonet] Fascin Hi all---one immuno question----is anyone using an IVD Fascin stain on their Hodgkins cases. My docs like this---but most of my sources are RUO. Any input is appreciated. Thanks- Deb VanEyck This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Tue Sep 13 11:23:23 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Arundo (Bamboo Cane Reeds) Message-ID: <20050913162323.96573.qmail@web50307.mail.yahoo.com> I am a histotech as well as an oboist. Like bassoons, they are a double reed instrument. Two pieces of bamboo are cut, shaved thin on one end, and tied together. Your bassoonists friends must be "pulling some all nighters" if their reeds are only lasting a week. As the clarinetist said, they need to be flexible, because the vibration of the reeds is what makes the sound. Many sites show how to construct reeds as she said also. I think some companies are making reeds from plastic that is safe to use, but the quality of the sound produced is not as good as natural bamboo. Some people use the plastic ones for practice and save the bamboo for performances. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From weneng2004 <@t> yahoo.com Tue Sep 13 11:56:29 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Thanks! Message-ID: <20050913165630.56989.qmail@web53414.mail.yahoo.com> Many thanks to all who adviced me on calcium stain! So far I know there are three stains to demonstrate calcium, Von Kossa, Alizarin red and Dahl's. I really appreciate the help I got from you! Wen --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From rmeyers <@t> ctcsurge.com Tue Sep 13 12:11:12 2005 From: rmeyers <@t> ctcsurge.com (Roger Meyers) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] automated response Message-ID: <10509131311.AA04497@mail.ctcsurge.com> I will be out of the office from Tuesday, September 13, 2005 through Wednesday, September 14, 2005. If the matter is urgent, please call Computer Trust Corporation at 617-557-9264 for immediate assistance. From sharon.osborn <@t> dnax.org Tue Sep 13 14:25:30 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] RE: slippy floors due to paraffin Message-ID: <29B25753F6B1D51196110002A589D444023981A7@PALMSG30.us.schp.com> Histonetters, One way that we discovered to cut down on the amount of paraffin on our floors was to change our method of trimming excess paraffin off the blocks before microtomy. In addition to changing our method, we also eliminated a major cause/stressor of repetitive motion syndrome. We had been using a knife to scrape off the paraffin from the sides and ends of the blocks after embedding and before microtomy so they would more easily fit the chuck. (and to keep the chuck from becoming loose so it would not well hold a block over time due to irregular sizing of blocks because of the paraffin on edges). We purchased the Thermo (Shandon) Electron Para Trimmer. Our paraffin particles on the floor has greatly reduced; our hands are not so tired from the manual scraping, etc. One caveat...you must remember to turn off the paratrimmer for it is a heated instrument and can burn. It is a life saver in reducing floor slippage and repetitive motion syndrome...outweighs our forgetting to turn it off..:-) sharon osborn DNAX, SP BioPharma Palo Alto, Ca ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From gagnone <@t> KGH.KARI.NET Tue Sep 13 14:26:04 2005 From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Giemsa bits... Message-ID: Hello Histonetters, We have an intermittent problem with our Giemsa stain for helicobacter. ( We are using EMD Giemsa stain catalogue # R03055/74.) The intermittent problem is a diamond-shaped deposit over the slide which is visible microscopically upon removal of slides from the warm solution. Not sure if it is reagent-related or not. We have tried to standardize all other factors i.e. pH, mechanical/technique variations among techs, staining time, filtering etc. Just wondering if anyone else out there has had the same problem. So far, pathologists are not complaining, as they can see around it, but it would be nice to eradicate it!! Thanks in advance, Eric From jcresor <@t> lcpath.com Tue Sep 13 14:32:03 2005 From: jcresor <@t> lcpath.com (Jennifer N. Cresor) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] blood marrow smears Message-ID: <200509131932.j8DJW8224988@plus34.host4u.net> Hello all, Has anyone out there heard of or had any experiences using acid alcohol to decolorize bone marrow smears for restaining? If so have you seen a loss of staining? Is this something that should be avoided? Any input would be appreciated. Jennifer jcresor@icpath.com From jkiernan <@t> uwo.ca Tue Sep 13 15:05:05 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] frozen section trichrome References: <20050909182556.18218.qmail@web90204.mail.scd.yahoo.com> Message-ID: <432730F1.CE232773@uwo.ca> Expect funny results when trichrome stains (devised for paraffin sections) are done on frozen sections. There have been a number of recent Histonet postings on this subject. See www.histosearch.com and search for "trichrome frozen" (without the quotation marks). In 1963 WK Engel & GG Cunningham (Neurology 13: 919-923) published a modified trichrome for cryostat sections of unfixed muscle (human biopsies and a wide variety of animals). It differs in important ways from its parent method (Gomori's one-step trichrome). My comments on the stages of the technique are indented. (I hope it looks OK after it's been emailed. If it's a real mess, please let me know.) There is no pre-staining treatment with picric acid or Bouin's fluid, as is usual before doing a regular trichrome on paraffin sections of formaldehyde-fixed material. 1. Unfixed sections stained 5 min in Harris's haemalum. In a sense, the tissue is also being fixed at this stage, because the stain contains a large excess of aluminium ions over haematein molecules, and it is quite acidic (pH about 2?). A solution of an aluminium salt doesn't work as a fixative for pieces of tissue (see Histochem. J. 17:1131-1146, 1986) but aluminium salts do make thin layers of gelatin insoluble in warm water, and are used as hardening agents for photographic films and printing papers. 2. Wash in distilled water X3. Note there was no acid-alcohol differentiation of the haemalum, and no blueing step with the washing. (There would be no point because the next step has pH less than 5, which can be expected to change the haemalum colour from blue to a dull, weary red.) 3. Stain for 10 min in Gomori's one-step trichrome solution that has been adjusted to pH 3.4 by adding 1N (4%) NaOH. For paraffin sections, there's published work to support lowering the pH of Gomori's mixture, to enhance the contrast between cytoplasm and thin collagen fibres. The anions of phosphotungstic acid (PTA) change into other anions (some larger and some smaller than PTA) in solutions with pH above 2. In most trichrome methods the pH of the solution containing PTA (or PMA) is less than 2. Engel & Cunningham's 1963 paper contains no explanation of the experiments that led them to pre-stain in Harris's haemalum, and there is no description of staining by Gomori-type trichrome solutions at pH other than 3.4. 4. A few dips in 0.2% acetic acid. The authors wrote "differentiate by a few dips in 0.2% acetic acid." Acidified water PREVENTS removal of anionic dyes, and is the opposite of differentiation in this technique. Engel & Cunningham (1963) did not advise checking wet slides under a microscope at any stage. 5. "Dehydrate and mount in Permount." The authors didn't give details. For trichrome methods I find it best to go straight from the acidified water into the first of three changes of 100% alcohol. (Alcohol-water mixtures can extract some of the bound dyes.) Unfortunately Engel & Cunningham (1963) did not give any account of the experiments that led to the development of their procedure. Neither did they discuss the possible mechanism of action, other than showing that the red staining of intermyofibrillar cytoplasm was prevented by strong lipid solvents, and therefore probably depended on intact mitochondria and sarcoplasmic reticulum. At pH 3.4 and preceded by 5 min in Harris's haemalum, the staining mechanisms must differ from those of regular trichromes, which are still controversial. See Prento 2001 Biotech. Histochem. 76:137-161 for some fairly recent discussion of the suject. John Kiernan Anatomy, UWO London, Canada. ______________________________________________________ Steven Coakley wrote: > > I'm looking for a consistant protocol for a trichrome stain for FS. I tried 2 this morning on liver. Both were sectioned at 7u , air dried for 30 minutes, fixed in bouins for 30 minutes and stained. > Extreamly red with a granular appearence. Only green was in the folds. My paraffin section control was fine. > > Steve > > > --------------------------------- > Click here to donate to the Hurricane Katrina relief effort. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.reilly <@t> jcu.edu.au Tue Sep 13 18:19:04 2005 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:25:33 2005 Subject: [Histonet] Image Analysis In-Reply-To: <20050913165630.56989.qmail@web53414.mail.yahoo.com> Message-ID: <5.1.0.14.0.20050914091337.00c3a628@mail.jcu.edu.au> >Hello Histonetters, Can anyone recommend an image analysis system which can measure the intensity of DAB staining in an immunohistochemical staining procedure? Thanks and regards, Laurie. Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From Xudong_Cao <@t> brown.edu Tue Sep 13 18:29:27 2005 From: Xudong_Cao <@t> brown.edu (Cao, Xudong) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] human origin cell marker Message-ID: <1DA88DDC48CCC245A777599FDAEED6D002C6036D@MAIL1.AD.Brown.Edu> Dear all, I am looking for a human origin cell marker to identify the harvested human skin (keratinocytes) grafted on nude mice. Would HLA-ABC be a good choice? thanks. Xudong From tahseen <@t> brain.net.pk Tue Sep 13 12:55:04 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Autolysis Specimen Message-ID: <003101c5b88c$46748220$972bfea9@m7c0y4> Does anyone have a good protocol for autolysis specimen. Muhammad Tahseen SKMCH&RC From M.Prideaux <@t> sheffield.ac.uk Wed Sep 14 03:44:25 2005 From: M.Prideaux <@t> sheffield.ac.uk (Matt Prideaux) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Automated microtomes In-Reply-To: <98E6E88F806B194A809BA1D62B9FA2CD2E7757@TXCLMAIL01.ameripath.local> References: <98E6E88F806B194A809BA1D62B9FA2CD2E7757@TXCLMAIL01.ameripath.local> Message-ID: <1126687465.4327e2e93c093@webmail.shef.ac.uk> We use a Leica RM2265 in our lab and have had no problems safety wise so far. So long as the microtome is being used carefully and the tech is fully trained there should be no problem. Personally, I find it great for when blocks require a lot of trimming and reduces the risk of RSI associated with the prolonged use of rotary microtomes. Hope that helps Matt -- Matt Prideaux Academic Unit of Bone Biology Division of Clinical Sciences (South) D Floor (DU20) Medical School Henry Wellcome Laboratories for Medical Research University of Sheffield Beech Hill Road Sheffield S10 2RX Tel: 0114 271 3783 Fax: 0114 271 1711 Quoting "Gomez, Milton" : > Dear Histonetters, > > What are your thoughts on injuries related to the use of automated > microtomes? I have heard that the use of automated microtomes increases the > risk of injury and in most cases these are more severe. May you share your > experience? I do know of one case where the tech lost part of his finger > (luckily he was able to have it re-attached and the finger was saved). How > many labs out there have used automated microtomes and have reverted back to > manual microtomes for safety reasons? Does safety take priority over > ergonomic reasons? > > Thanks in advance for your reply, > Milton > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Kemlo.Rogerson <@t> elht.nhs.uk Wed Sep 14 04:11:07 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Automated microtomes Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698FCE@elht-exch1.xelht.nhs.uk> Can you get an automated microscope? My neck hurts? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Matt Prideaux Sent: 14 September 2005 09:44 To: Gomez, Milton; Histonet Subject: Re: [Histonet] Automated microtomes We use a Leica RM2265 in our lab and have had no problems safety wise so far. So long as the microtome is being used carefully and the tech is fully trained there should be no problem. Personally, I find it great for when blocks require a lot of trimming and reduces the risk of RSI associated with the prolonged use of rotary microtomes. Hope that helps Matt -- Matt Prideaux Academic Unit of Bone Biology Division of Clinical Sciences (South) D Floor (DU20) Medical School Henry Wellcome Laboratories for Medical Research University of Sheffield Beech Hill Road Sheffield S10 2RX Tel: 0114 271 3783 Fax: 0114 271 1711 Quoting "Gomez, Milton" : > Dear Histonetters, > > What are your thoughts on injuries related to the use of automated > microtomes? I have heard that the use of automated microtomes increases the > risk of injury and in most cases these are more severe. May you share your > experience? I do know of one case where the tech lost part of his finger > (luckily he was able to have it re-attached and the finger was saved). How > many labs out there have used automated microtomes and have reverted back to > manual microtomes for safety reasons? Does safety take priority over > ergonomic reasons? > > Thanks in advance for your reply, > Milton > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Sep 14 06:16:00 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Re: Calcium staining Message-ID: Von Kossa is one we have used in the past- Reagents: 0.1M citrate buffer pH 4.5: 0.2M disodium hydrogen phosphate - 9.09ml 0.1M citric acid - 10.91ml 5% silver nitrate 5% sodium thiosulphate Method: Take 2 sections to water Immerse 1 in citrate buffer - 20 mins for neg control Wash both slides and a coplin jar well in dist water Put slides in coplin jar and fill with 5% silver nitrate Expose to bright sunlight-10 to 20 mins or (if you are in England) a 60 watt bulb at range of 4-5 inches - 30-60 mins Wash both slides well in dist water Treat with 5% sodium thiosulphate - 2-3 mins Counterstain Dehydrate, clear and mount Alizarin red is also a good method if you have the dye. Jacqui Malam Royal Infirmary Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From failm <@t> musc.edu Wed Sep 14 07:32:56 2005 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Giemsa bits... Message-ID: Have you triede filterin the solution after microwaving? >>> "Gagnon, Eric" 09/13/05 15:33 PM >>> Hello Histonetters, We have an intermittent problem with our Giemsa stain for helicobacter. ( We are using EMD Giemsa stain catalogue # R03055/74.) The intermittent problem is a diamond-shaped deposit over the slide which is visible microscopically upon removal of slides from the warm solution. Not sure if it is reagent-related or not. We have tried to standardize all other factors i.e. pH, mechanical/technique variations among techs, staining time, filtering etc. Just wondering if anyone else out there has had the same problem. So far, pathologists are not complaining, as they can see around it, but it would be nice to eradicate it!! Thanks in advance, Eric _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From failm <@t> musc.edu Wed Sep 14 07:36:01 2005 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Giemsa bits... Message-ID: Sorry, A few misspellings in the first e-mail. Try filtering the giemsa after microwaving. Rena Fail >>> "Gagnon, Eric" 09/13/05 15:33 PM >>> Hello Histonetters, We have an intermittent problem with our Giemsa stain for helicobacter. ( We are using EMD Giemsa stain catalogue # R03055/74.) The intermittent problem is a diamond-shaped deposit over the slide which is visible microscopically upon removal of slides from the warm solution. Not sure if it is reagent-related or not. We have tried to standardize all other factors i.e. pH, mechanical/technique variations among techs, staining time, filtering etc. Just wondering if anyone else out there has had the same problem. So far, pathologists are not complaining, as they can see around it, but it would be nice to eradicate it!! Thanks in advance, Eric _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NHeath <@t> Lifespan.org Wed Sep 14 08:17:23 2005 From: NHeath <@t> Lifespan.org (Heath, Nancy L.) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] frozen section trichrome Message-ID: <09C945920A6B654199F7A58A1D7D1FDE65ECF8@lsexch.lsmaster.lifespan.org> This is for Steven Coakley... I could email you my Trichrome procedure I use here on frozen muscle. Nancy Heath, HT(ASCP) Neuropath Dept. Rhode Island Hospital -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of John Kiernan Sent: Tuesday, September 13, 2005 4:05 PM To: Steven Coakley; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] frozen section trichrome Expect funny results when trichrome stains (devised for paraffin sections) are done on frozen sections. There have been a number of recent Histonet postings on this subject. See www.histosearch.com and search for "trichrome frozen" (without the quotation marks). In 1963 WK Engel & GG Cunningham (Neurology 13: 919-923) published a modified trichrome for cryostat sections of unfixed muscle (human biopsies and a wide variety of animals). It differs in important ways from its parent method (Gomori's one-step trichrome). My comments on the stages of the technique are indented. (I hope it looks OK after it's been emailed. If it's a real mess, please let me know.) There is no pre-staining treatment with picric acid or Bouin's fluid, as is usual before doing a regular trichrome on paraffin sections of formaldehyde-fixed material. 1. Unfixed sections stained 5 min in Harris's haemalum. In a sense, the tissue is also being fixed at this stage, because the stain contains a large excess of aluminium ions over haematein molecules, and it is quite acidic (pH about 2?). A solution of an aluminium salt doesn't work as a fixative for pieces of tissue (see Histochem. J. 17:1131-1146, 1986) but aluminium salts do make thin layers of gelatin insoluble in warm water, and are used as hardening agents for photographic films and printing papers. 2. Wash in distilled water X3. Note there was no acid-alcohol differentiation of the haemalum, and no blueing step with the washing. (There would be no point because the next step has pH less than 5, which can be expected to change the haemalum colour from blue to a dull, weary red.) 3. Stain for 10 min in Gomori's one-step trichrome solution that has been adjusted to pH 3.4 by adding 1N (4%) NaOH. For paraffin sections, there's published work to support lowering the pH of Gomori's mixture, to enhance the contrast between cytoplasm and thin collagen fibres. The anions of phosphotungstic acid (PTA) change into other anions (some larger and some smaller than PTA) in solutions with pH above 2. In most trichrome methods the pH of the solution containing PTA (or PMA) is less than 2. Engel & Cunningham's 1963 paper contains no explanation of the experiments that led them to pre-stain in Harris's haemalum, and there is no description of staining by Gomori-type trichrome solutions at pH other than 3.4. 4. A few dips in 0.2% acetic acid. The authors wrote "differentiate by a few dips in 0.2% acetic acid." Acidified water PREVENTS removal of anionic dyes, and is the opposite of differentiation in this technique. Engel & Cunningham (1963) did not advise checking wet slides under a microscope at any stage. 5. "Dehydrate and mount in Permount." The authors didn't give details. For trichrome methods I find it best to go straight from the acidified water into the first of three changes of 100% alcohol. (Alcohol-water mixtures can extract some of the bound dyes.) Unfortunately Engel & Cunningham (1963) did not give any account of the experiments that led to the development of their procedure. Neither did they discuss the possible mechanism of action, other than showing that the red staining of intermyofibrillar cytoplasm was prevented by strong lipid solvents, and therefore probably depended on intact mitochondria and sarcoplasmic reticulum. At pH 3.4 and preceded by 5 min in Harris's haemalum, the staining mechanisms must differ from those of regular trichromes, which are still controversial. See Prento 2001 Biotech. Histochem. 76:137-161 for some fairly recent discussion of the suject. John Kiernan Anatomy, UWO London, Canada. ______________________________________________________ Steven Coakley wrote: > > I'm looking for a consistant protocol for a trichrome stain for FS. I tried 2 this morning on liver. Both were sectioned at 7u , air dried for 30 minutes, fixed in bouins for 30 minutes and stained. > Extreamly red with a granular appearence. Only green was in the folds. My paraffin section control was fine. > > Steve > > > --------------------------------- > Click here to donate to the Hurricane Katrina relief effort. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Sep 14 10:28:57 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Image Analysis References: <5.1.0.14.0.20050914091337.00c3a628@mail.jcu.edu.au> Message-ID: <432841B9.DA11273@uwo.ca> The intensity of DAB staining does not usually indicate the quantity or concentration of an antigen in an immunoperoxidase section. There are even circumstances in which the amount of oxidized DAB can vary inversely with the local concentration of the antigen. In general: enzyme amplification can detect and localize very small amounts of some antigens, but the intensity of staining has no quantitative significance. There is a large body of peer-reviewed literature in the field of quantitative immunohistochemistry. Optical measurements correlate with antigen concentration only with the simplest methods. John Kiernan Anatomy Dept, UWO London, Canada. _________________________ Laurie Reilly wrote: > > >Hello Histonetters, > > Can anyone recommend an image analysis system which can measure the > intensity of DAB staining in an > immunohistochemical staining procedure? > > Thanks and regards, Laurie. > > Mr.Laurie Reilly Ph 07 4781 4468 > School of Veterinary & Biomedical Science Fax 07 4779 1526 > James Cook University > Townsville Qld. > 4811 laurie.reilly@jcu.edu.au > > Australia. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From drgrant <@t> cmh.edu Wed Sep 14 12:51:11 2005 From: drgrant <@t> cmh.edu (Grant, Debra, R) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] IHC Forms Message-ID: <6F434E27D5DE944B9E898984CADDD14904392C@exchmail.CMH.Internal> Hi all, We are currently working up a new Immunohistochemistry (IHC) work up sheet for new antibodies for the pathologists to sign off on. Anyone willing to share examples of their forms please e-mail them directly to me. Debby R. Grant HT(ASCP) Histology Department The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 drgrant@cmh.edu 816-234-3827 From RSRICHMOND <@t> aol.com Wed Sep 14 14:28:59 2005 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Re: frozen section trichrome Message-ID: <84.4da2692e.3059d3fb@aol.com> I'm glad John Kiernan had the formula for the Engel-Cunningham frozen section trichrome stain on hand. I tried to set up that stain for human skeletal muscle frozen sections in 1969, when I was a fellow in the pathology department at Johns Hopkins. I was unable to get the stain to work, and I finally visited Guy Cunningham, the histotechnologist in W. King Engel's muscle pathology laboratory at the National Institutes of Health. They let me look at their procedure in detail. I noted that they were using a true Harris hematoxylin - oxidized with mercuric oxide - while we were using an iodate-oxidized hematoxylin, home-made (John R. Baker's formula) in our research lab. I quickly purloined some Harris hematoxylin from the cytopathology service, and after that the stain worked fine. I added mercuric chloride to our standard hematoxylin, and that worked too. That was 35 years ago. You couldn't buy hematoxylin with mercury in it nowadays, and you wouldn't want to make it either. There are still commercial hematoxylins labeled Harris hematoxylin, which I think now means "any alum hematoxylin our marketing department wants to call Harris hematoxylin". I'm curious to know what's become of this problem. Bob Richmond former Samurai Pathologist now at Gaston Memorial Hospital, Gastonia NC From Robert.Lott <@t> bhsala.com Wed Sep 14 14:38:59 2005 From: Robert.Lott <@t> bhsala.com (Lott, Robert) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Giemsa bits.... Message-ID: The problem you are experiencing is a common one with commercial Giemsa solutions. Upon heating them a precipitate forms in the working solution(s). The solution to this problem was taught to me by one of the masters of our craft... Charles Churukian, Supervisor of the Special Stains Laboratory, University of Rochester Medical Center. His recommendation is to add about 0.5 - 1.0 ml of 10% Triton X-100 (a common laboratory detergent) to the working solution(s) "before" heating them. The addition of the detergent results in "more distinct staining" of the various cell types as well as acting as a stabilizer when heating the Giemsa solution(s). This apparent heating phenomenon does not occur if the solutions are used at room temperature. Try it and see how it works!!! Robert L. Lott, HTL(ASCP) Manager, Anatomic Pathology Baptist Health System 800 Montclair Road Birmingham, AL 35213 205-592-5388 phone 205-592-5646 fax robert.lott@bhsala.com ------------------------------------------------------------------------ ------------------------------------------------------------------------ ------------------------------------------------------------------------ ----- Date: Tue, 13 Sep 2005 15:26:04 -0400 From: "Gagnon, Eric" Subject: [Histonet] Giemsa bits... To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello Histonetters, We have an intermittent problem with our Giemsa stain for helicobacter. ( We are using EMD Giemsa stain catalogue # R03055/74.) The intermittent problem is a diamond-shaped deposit over the slide which is visible microscopically upon removal of slides from the warm solution. Not sure if it is reagent-related or not. We have tried to standardize all other factors i.e. pH, mechanical/technique variations among techs, staining time, filtering etc. Just wondering if anyone else out there has had the same problem. So far, pathologists are not complaining, as they can see around it, but it would be nice to eradicate it!! Thanks in advance, Eric Confidentiality Notice: The information contained in this email message is privileged and confidential information and intended only for the use of the individual or entity named in the address. If you are not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this information is strictly prohibited. If you received this information in error, please notify the sender and delete this information from your computer and retain no copies of any of this information. From deb.vaneyck <@t> phci.org Wed Sep 14 16:37:46 2005 From: deb.vaneyck <@t> phci.org (Van Eyck, Deb) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] CD 20 on Bouins fizxed tissue Message-ID: Does anyone use a cd 20 antibody which works well on bouins fixed tissue??? Since b-5 is gone we are looking at other fixatives for optimal morphology and staining in some of our CD markers-----so any input would be great! Thanks- Deb This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. From cfavara <@t> niaid.nih.gov Wed Sep 14 17:46:08 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] recycling Message-ID: I did not get much response for recycling dehydrating agents and solvents. Please if you recycle these let me hear form you! Thanks,. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From fredericoacazevedo <@t> gmail.com Wed Sep 14 10:05:44 2005 From: fredericoacazevedo <@t> gmail.com (frederico azevedo) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Need manual for crysostat Cryocut 1800 Message-ID: Hi everyone, I need the manual of the cryostat Cryocut 1800 (Leica). I already tried to search at Leica?s home page but did not find anything. Anyone knows how to help me? Thanks, Frederico A. C. Azevedo Dept. of Anatomy - Neuroplasticity UFRJ, Rio de Janeiro, Brazil. From jim.manavis <@t> imvs.sa.gov.au Thu Sep 15 02:25:41 2005 From: jim.manavis <@t> imvs.sa.gov.au (Jim Manavis) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] D1 and D2 antibodies Message-ID: <000001c5b9c6$ada015a0$636c140a@itp36533> Has anyone experience with the Dopamine D1 and D2 receptor antibodies in formalin fixed paraffin embedded brain tissue. Cheers Jim From hilsley <@t> capeheart.uct.ac.za Thu Sep 15 03:00:12 2005 From: hilsley <@t> capeheart.uct.ac.za (Helen Ilsley) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] sudan black/bleaching Message-ID: -- Hi I wonder if any one can help me. I am using sudan black to suppress my backgound autofluorescence but when I want to photograph my section I seem to be getting photobleaching especially at high mags. The image under the microscope blurs and when I move away from the area I am left with a bleached, blurry ring. Is this due to the sudan black? Has anyone ever had this sort of problem before. I would really appreciate any help. Thanks Helen Ilsley Chris Barnard Division of Cardiothoracic Surgery Cape Heart Centre University of Cape Town Medical School Cape Town, South Africa From Andrew.MacDuff <@t> ed.ac.uk Thu Sep 15 08:08:07 2005 From: Andrew.MacDuff <@t> ed.ac.uk (Andrew MacDuff) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Mouse neutrophil isolation Message-ID: <000a01c5b9f6$84625f60$93eed781@universi8lynpq> Hi I'm wanting to isolate mouse neutrophils from peripheral blood. There are a number of refs in the journals to a kit from a company called Cardinal Associates in Santa Fe, NM but I can't get in touch with them. Does anyone have conact details or know if they've merged/gone bust? If anyone has a protocol for mouse PMN isolation which they could let me have that would be great too. Thanks Andrew Andrew MacDuff Clinical Research Fellow Medical School Edinburgh University Teviot Place Edinburgh andrew.macduff@ed.ac.uk From gagnone <@t> KGH.KARI.NET Thu Sep 15 08:33:25 2005 From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Giemsa Solutions Message-ID: Rena and Robert, thanks for your input, we are trying your suggestions and hopefully will meet with success. Your help is appreciated. Thanks, Eric From vazquezr <@t> ohsu.edu Thu Sep 15 09:04:32 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Need manual for crysostat Cryocut 1800 Message-ID: I purchased mine from the manufacture. It's not that expensive. robyn ohsu >>> frederico azevedo 9/14/2005 8:05 AM >>> Hi everyone, I need the manual of the cryostat Cryocut 1800 (Leica). I already tried to search at Leica?s home page but did not find anything. Anyone knows how to help me? Thanks, Frederico A. C. Azevedo Dept. of Anatomy - Neuroplasticity UFRJ, Rio de Janeiro, Brazil. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jluis.palazon <@t> icman.csic.es Thu Sep 15 09:46:57 2005 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] antibodies Message-ID: <20050915144657.BACD38F8C06@perceval.uca.es> Dear List-members Could any of you recomend me what would be the best antibody for the identification of smooth muscle cells in fish? anti-alfa miosin? thanks in advance for the advice Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From ali.moussavi <@t> molbio.gu.se Thu Sep 15 10:13:55 2005 From: ali.moussavi <@t> molbio.gu.se (Ali Moussavi Nik) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] question about the embryonic brain fixation Message-ID: <94E1697CA3@mailer.lundberg.gu.se> Dear All I would like to section some mouse embryonic brain (18.5 days) with vibrotom. I will do some Immunocytochemistry on them .I noticed that perfusion of the mother is the best way for fixation of the embryo's brain. but which fixative makes the lowest background? Could some one give me a good protocol for that. Thanks in advance. Ali Ali Moussavi Nik D.V.M, PhD student Department of Cell and Molecular Biology,Lundburg lab, Gothenburg,Sweden From sjchtascp <@t> yahoo.com Thu Sep 15 13:37:02 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] sucrose/fixation Message-ID: <20050915183702.32134.qmail@web90202.mail.scd.yahoo.com> I work in research and am looking for as much information on sucrose or other tissue cryoprotection as possible. Our scientist is not pleased with the tissue morphology, usually prefixed at 4C. So information on times, temp of fixation, type of sutible fixatives. This is mainly for IF. Steve --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From akbitting <@t> geisinger.edu Thu Sep 15 14:36:31 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] The Histology Lab at Geisinger Medical Center, a large physician run hospital in Danville (Montour C Message-ID: The Histology Lab at Geisinger Medical Center, a large physician run hospital in Danville (Montour County), PA has an immediate FT opening for a Histotechnician/Histotechnologist, registered or registry eligible. No weekends or holidays. Industry competitive compensation & benefits package. Email me at akbitting@geisinger.edu or call me at 570-214-9634 for information. You can also apply at our website www.geisinger.edu Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Sep 16 02:38:45 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] RE: Antibody work up sheet Message-ID: I wouldn't mind seeing some examples if anyone has some to share. Thanks Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From PKamalavenkatesh <@t> wockhardtin.com Fri Sep 16 05:30:32 2005 From: PKamalavenkatesh <@t> wockhardtin.com (PKamalavenkatesh@wockhardtin.com) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Ornithine carbamyl transferase- histochemistry- Rat liver Message-ID: Dear all, This is Dr.Kamalavenkatesh from wockhardt reserch center, India. We want to do quantitative histochemistry of Ornithine carbamyl transferase (OCT) in rat liver. We have searched google and also pubmed for the protocols or research articles regarding the methodology. It seems that the information regarding this subject is scanty. Hence, i want to know from you that any body is doing this technique in their laboratory, if so the protocol involved and their experience with the method. Please let us know. Thanks and regards kamalavenkatesh --------------------- Disclaimer ------------------------------------------------------------------------------ Information transmitted by this E-MAIL is proprietary to Wockhardt Ltd. and/ or its Group Companies and/or,its Customers and is intended for use only by the individual or entity to which it is addressed, and may contain information that is privileged, confidential or exempt from disclosure under applicable law. If you are not the intended recipient or it appears that this mail has been forwarded to you without proper authority, you are notified that any use or dissemination of this information in any manner is strictly prohibited. In such cases, please delete this mail from your records. --------------------------------------------------------------------------------------------------------------- From Sue.Kapoor <@t> uhsi.org Fri Sep 16 08:46:16 2005 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] CK5/6 questions Message-ID: <61E9F2400F53D5119CFC00508B44E33B02E9E194@khmcexch.uhsi.org> Hello everyone, My doc just came back from a conference and has some questions pertaining to CK 5/6.... 1. Are folks finding better results using CK 5/6 on cellblock preparations over using TTF-1 for mesothelioma? He was told that TTF-1 is adversely affected by 95% alcohol fixation, does anyone know of this? 2. Is anyone finding better sensitivity for prostate basal cells over HMW-CK (34betaE12)? Much thanks in advance! Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From billingconsultants <@t> yahoo.com Fri Sep 16 08:49:19 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] MLH1, MSH2 controls Message-ID: <20050916134920.76202.qmail@web54206.mail.yahoo.com> Hi everyone, I was wondering if any of you might know of a vendor or have a suggestion for MLH1 and MSH2 controls. Thank you in advance for your assistance. Louri __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From petepath <@t> yahoo.com Fri Sep 16 09:14:09 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] fine needle aspirates assisting Message-ID: <20050916141410.12148.qmail@web30412.mail.mud.yahoo.com> Reviewing the posts I missed while at NSH I thought I could offer a bit on this one. At our hospital we have the cytotechs go up first on most FNA's and smear, stain and screen the slides. They call us when they get a positive or if the radiologist asks to bring a pathologist to check the slides rather than continuing to stick the patient. We then walk over and read the slide. This saves us a great deal of time. I will often go from the beginning if it is something I know will be difficult like pancreas or lymphoma cases. Our cart has H&E and diff quick stains and keeps RPMI and formalin for flow and cell blocks. Here are a few things that some of you may find useful. 1) When I make smears rather than put a drop on each slide and smear them, I blow it all on one slide and use an additional slide to pick up an aliquot and smear the material to additional slides. This can be done very quickly before things begin clot too much. It also gives you a mirror image of the material on the slides so if you are comparing diff quick to h&e it will have a similar population. When the material clots it will not smear thin and is considerably less readable. Work as fast as you can without sticking yourself. 2) If you get a bloody pass, quickly blow it all out on one slide, tilt it up and let the blood run onto a second slide. The specks of tissue will adhere to the first slide. Pick the tissue specks a few at a time up using a third slide and smear them onto additional slides. These will make very readable blood free slides. Let the puddle of blood that you initially ran off clot and use it for cell block. 3) If I read the final positive slides and I sence that I will need immunos to make a definitive diagnosis, I will tell the radiologist to give me another aggresive bloody pass. We blow it out and let it clot and use this for cell block. Same goes if I think I need flow, I will ask for a pass to put in RPMI. 4) I continue to ask for passes until I have a good sample or the radiologist gives up. If you appear to have less than diagnostic material when you leave radiology, it usually does not magically turn positive in your lab. Sometimes the cell block will have something that you did not see on the smear but don't count on it. And by good material I mean an nice cellular slide. If the radiologist puts a neede into tumor it should yield numerous cells and clusters. The exception is very sclerotic lesions such as an old schwannoma where you will be lucky to get a few small fragments back. Stephen From Rcartun <@t> harthosp.org Fri Sep 16 09:21:33 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] CK5/6 questions Message-ID: Hi Sue: Regarding: 1. In general, CK5/6 (squamous cell CA & mesothelioma) and TTF-1 (pulmonary adenocarcinoma) stain different tumors. Although I agree that alcohol fixation can be detrimental to certain proteins, I have had good luck with TTF-1 on cell blocks and alcohol-fixed direct cytology smears. I like calretinin (Zymed polyclonal) for mesothelial cells. Recently, a new marker "D2-40" has been used for mesothelioma with excellent results. You may want to try CK5 from Novocastra; I like it better than CK5/6. 2. We only use high molecular weight cytokeratin (clone 34BetaE12) for identifying basal cells in prostatic tissue. Occasionally, we may order P504S. I know many labs are routinely running multiple antibodies (even cocktails) for basal cells, but, in my opinion, this is unnecessary and only makes healthcare more expensive. However, many labs may be getting suboptimal results for HMW-CK and this has forced them to use other basal cell markers. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Kapoor, Sue" 09/16/05 09:46AM >>> Hello everyone, My doc just came back from a conference and has some questions pertaining to CK 5/6.... 1. Are folks finding better results using CK 5/6 on cellblock preparations over using TTF-1 for mesothelioma? He was told that TTF-1 is adversely affected by 95% alcohol fixation, does anyone know of this? 2. Is anyone finding better sensitivity for prostate basal cells over HMW-CK (34betaE12)? Much thanks in advance! Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From fredericoacazevedo <@t> gmail.com Fri Sep 16 09:59:24 2005 From: fredericoacazevedo <@t> gmail.com (frederico azevedo) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Thanks! Copy manual Cryocut 1800 obtained! Message-ID: Thank you everyone! Mr. Monfils will send me a copy of the manual by mail. I appreciate the efforts of you all. I?ve got lots of answers trying to solve my problem. I?m really glad! Frederico A. C. Azevedo Neuroanatomy dept., UFRJ Brasil. From RSRICHMOND <@t> aol.com Fri Sep 16 11:06:39 2005 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] heart biopsy Message-ID: <15b.5967d6b8.305c478f@aol.com> Our pathology department is about to receive its first heart (endomyocardial) biopsy - being done to look for amyloid, not a transplant - What protocol do you all have for these specimens? One service I spoke with does two H & E, trichrome, iron, and amyloid - on formalin fixed tissue. What additionally is needed for transplant heart biopsy specimens? Bob Richmond Gaston Memorial Hospital, Gastonia NC From lrichey <@t> u.washington.edu Fri Sep 16 12:05:29 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] heart biopsy In-Reply-To: <15b.5967d6b8.305c478f@aol.com> References: <15b.5967d6b8.305c478f@aol.com> Message-ID: <432AFB59.6040602@u.washington.edu> We do 5 levels with an H&E and 3 unst at each level. IHC is done on the unst if needed. Congo Red is cut at 8 microns. RSRICHMOND@aol.com wrote: >Our pathology department is about to receive its first heart (endomyocardial) >biopsy - being done to look for amyloid, not a transplant - > >What protocol do you all have for these specimens? One service I spoke with >does two H & E, trichrome, iron, and amyloid - on formalin fixed tissue. > >What additionally is needed for transplant heart biopsy specimens? > >Bob Richmond >Gaston Memorial Hospital, Gastonia NC >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From kerry.l.crabb <@t> gsk.com Fri Sep 16 12:38:34 2005 From: kerry.l.crabb <@t> gsk.com (kerry.l.crabb@gsk.com) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Kerry L Crabb/PharmRD/GSK is out of the office. Message-ID: I will be out of the office starting 07-Sep-2005 and will not return until 19-Sep-2005. During my absence contact Eve about necropsy issues and contact Teresa about histology issues. I will respond to messages when I return. From bcfinlay <@t> cox.net Fri Sep 16 13:15:46 2005 From: bcfinlay <@t> cox.net (bcfinlay@cox.net) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Northwest Florida Histology Full Time Position Message-ID: <20050916181538.JHTR29674.eastrmmtao03.cox.net@smtp.east.cox.net> Histology Technician or Technologist needed in Pensacola, Florida. Must be ASCP certified or eligible. The position is in a private outpatient lab, M-F with no weekends and no call. Please send resume and salary requirements to: Renae Taylor, PHR / Medical Center Clinic 8333 N. Davis Highway Pensacola, FL 32514 Fax - (850) 969-2963 email - careers@medicalcenterclinic.com From mucram11 <@t> comcast.net Fri Sep 16 13:17:50 2005 From: mucram11 <@t> comcast.net (pam marcum) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] New ASCP Classification and You Message-ID: <432B0C4E.00000E.02344@YOUR-4DI1S53IME> Good Morning, I have just returned from the NSH meeting. I was surprised to find out the Histology Assistant or HA category recently presented to the NSH board and refused with a letter is moving forward. No one had told the general membership of NSH or ASCP or had it been explained to them in full. It is important for those of us who have worked so hard to get the new the education rules set in place to understand what this new registry will be and how it will affect us as professionals. This is an aid position that is OJT (on the job training) and will mean the pathologist, hospital or other private histology laboratory will be able to avoid having more than registered HT or HTL in the laboratory. The fully registered HT or HTL can then be used as the supervisor or manager and responsible for all the HA?s work. This will aid in maintaining the lower pay scale we currently enjoy and help prevent us raising our overall image as professionals. Please find out more about this and respond to the ASCP/BOR with comments about this new category. The only way any of us will be heard is to let ASCP know what we think and I hope many of you will join me in letting them know we do not want to go back to the 1960s and 70s when we were all OJT and the pay is still reflecting it. (By the way I was OJT and have worked hard to improve my skills since then) We have worked hard for over thirty years to begin to be recognized as professionals not just techs! DO NOT let this stop us now so the pathologists at ASCP and AMA can continue to treat Histology as minor part of the laboratory. Histology has changed and will continue to make strides forward that require more education not less. The board of NSH did send a letter to the ASCP and tell them they would not approve this category. It is a personal feeling however, I think the Board of NSH should have notified the membership about this immediately, as a letter is not sufficient to protect us or respond as members of both NSH and ASCP. Again, write ASCP/BOR about your feelings if this important to you. Second point Cathy Locallo put a motion forward at the House of Delegates to request a Task Force to see if there is way to get histologists (HT and HTL) working for the federal government recognized as professionals and here we have a new way with the HA registry to keep us status quo. By the time the task force is in place and moving we will be down a rung again. Pamela Marcum (This is a reflection of my opinion not my employer or any other person.) Pam Marcum From pathrm35 <@t> adelphia.net Fri Sep 16 14:17:50 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:25:34 2005 Subject: [Histonet] Histotechs displaced by Katrina Message-ID: <30076697.1126898270454.JavaMail.root@web10.mail.adelphia.net> Fellow Techs, We are currently seeking a full time Histologist in Southeast Florida in a small but growing private dermpath lab. ASCP and Florida license or eligible required. My physician has informed me that if there are any displaced Histologists from Katrina who are interested, we will assist with relocation and housing. There is a sign on bonus for anyone else who is interested. Thank you, Ron Martin Benchmark Pathology Laboratory Fax 561-721-1249 From KMB1904 <@t> aol.com Fri Sep 16 15:16:36 2005 From: KMB1904 <@t> aol.com (KMB1904@aol.com) Date: Fri Sep 16 16:09:15 2005 Subject: [Histonet] JCAHO question Message-ID: When it comes to fume hoods ...what is the JCAHO regulation on fume hoods. We have coverslipping areas and staining areas that are not ventilated. I would love to be able to tell my supervisor where this is stated in the regulations. Thanks in advance for any advice! From Jackie.O'Connor <@t> abbott.com Fri Sep 16 16:59:26 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 17:00:02 2005 Subject: [Histonet] JCAHO question Message-ID: I think this is more of an OSHA violation. If you're staining and coverslipping in an area where xylene is used, and you don't have a fume hood, that's a violation of CFR 1910. Jackie O' KMB1904@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 09/16/2005 03:16 PM To: histonet@pathology.swmed.edu cc: Subject: [Histonet] JCAHO question When it comes to fume hoods ...what is the JCAHO regulation on fume hoods. We have coverslipping areas and staining areas that are not ventilated. I would love to be able to tell my supervisor where this is stated in the regulations. Thanks in advance for any advice! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Fri Sep 16 17:34:07 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 17:31:43 2005 Subject: [Histonet] it's. it's a stone Message-ID: <009c01c5bb0e$c05b91d0$52bf0b43@yourxhtr8hvc4p> For those of you who saw me agonize my way through the NSH this week (as well as my lecture). I have good news for you. The kidney stone was born Thursday, Sept 15 at 8:30 AM. I made it back to San Antonio without bringing down another plane. I saw my primary care physician and a urologist. I am mending, but can not take any plane rides soon. This means that I'll have to put off taking the PA exam until 2006, which is okay. Thank you all for you sympathy, prayers and well wishes. I shall return Joe " The Toe" Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX From jnocito <@t> satx.rr.com Fri Sep 16 17:36:13 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 17:33:43 2005 Subject: [Histonet] New ASCP Classification and You References: <432B0C4E.00000E.02344@YOUR-4DI1S53IME> Message-ID: <00a001c5bb0f$0b1fbca0$52bf0b43@yourxhtr8hvc4p> Why do I feel like we're doing the Texas Two Step here? Let's support Pam in this and as soon as I'm able to, I also will pick up my sword in defense. Besides, y'all know how I like to stir things up Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "pam marcum" To: Sent: Friday, September 16, 2005 1:17 PM Subject: [Histonet] New ASCP Classification and You > > > Good Morning, > > > > I have just returned from the NSH meeting. I was surprised to find out the > Histology Assistant or HA category recently presented to the NSH board and > refused with a letter is moving forward. No one had told the general > membership of NSH or ASCP or had it been explained to them in full. > > > > It is important for those of us who have worked so hard to get the new the > education rules set in place to understand what this new registry will be > and how it will affect us as professionals. This is an aid position that > is > OJT (on the job training) and will mean the pathologist, hospital or other > private histology laboratory will be able to avoid having more than > registered HT or HTL in the laboratory. The fully registered HT or HTL can > then be used as the supervisor or manager and responsible for all the HA?s > work. This will aid in maintaining the lower pay scale we currently enjoy > and help prevent us raising our overall image as professionals. > > > > Please find out more about this and respond to the ASCP/BOR with comments > about this new category. The only way any of us will be heard is to let > ASCP > know what we think and I hope many of you will join me in letting them > know > we do not want to go back to the 1960s and 70s when we were all OJT and > the > pay is still reflecting it. (By the way I was OJT and have worked hard to > improve my skills since then) We have worked hard for over thirty years to > begin to be recognized as professionals not just techs! DO NOT let this > stop > us now so the pathologists at ASCP and AMA can continue to treat Histology > as minor part of the laboratory. Histology has changed and will continue > to > make strides forward that require more education not less. > > > > The board of NSH did send a letter to the ASCP and tell them they would > not > approve this category. It is a personal feeling however, I think the Board > of NSH should have notified the membership about this immediately, as a > letter is not sufficient to protect us or respond as members of both NSH > and > ASCP. Again, write ASCP/BOR about your feelings if this important to you. > > > > Second point Cathy Locallo put a motion forward at the House of Delegates > to > request a Task Force to see if there is way to get histologists (HT and > HTL) > working for the federal government recognized as professionals and here we > have a new way with the HA registry to keep us status quo. By the time the > task force is in place and moving we will be down a rung again. > > > > Pamela Marcum > > (This is a reflection of my opinion not my employer or any other person.) > > > > > > Pam Marcum > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.344 / Virus Database: 267.11.1/104 - Release Date: 9/16/2005 > > From JWEEMS <@t> sjha.org Fri Sep 16 17:47:43 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 17:47:56 2005 Subject: [Histonet] JCAHO question Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01304C90@sjhaexc02.sjha.org> I would be more concerned about OSHA. They could shut the lab down for this violation! Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of KMB1904@aol.com Sent: Friday, September 16, 2005 4:17 PM To: histonet@pathology.swmed.edu Subject: [Histonet] JCAHO question When it comes to fume hoods ...what is the JCAHO regulation on fume hoods. We have coverslipping areas and staining areas that are not ventilated. I would love to be able to tell my supervisor where this is stated in the regulations. Thanks in advance for any advice! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From JWEEMS <@t> sjha.org Fri Sep 16 18:57:36 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 18:57:52 2005 Subject: [Histonet] New ASCP Classification and You Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01304C93@sjhaexc02.sjha.org> This is a shock! I don't recall ever hearing of this before. Where did it originate? Thanks for the information. And did everyone have a great time at NSH? Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of pam marcum Sent: Friday, September 16, 2005 2:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You Good Morning, I have just returned from the NSH meeting. I was surprised to find out the Histology Assistant or HA category recently presented to the NSH board and refused with a letter is moving forward. No one had told the general membership of NSH or ASCP or had it been explained to them in full. It is important for those of us who have worked so hard to get the new the education rules set in place to understand what this new registry will be and how it will affect us as professionals. This is an aid position that is OJT (on the job training) and will mean the pathologist, hospital or other private histology laboratory will be able to avoid having more than registered HT or HTL in the laboratory. The fully registered HT or HTL can then be used as the supervisor or manager and responsible for all the HA?s work. This will aid in maintaining the lower pay scale we currently enjoy and help prevent us raising our overall image as professionals. Please find out more about this and respond to the ASCP/BOR with comments about this new category. The only way any of us will be heard is to let ASCP know what we think and I hope many of you will join me in letting them know we do not want to go back to the 1960s and 70s when we were all OJT and the pay is still reflecting it. (By the way I was OJT and have worked hard to improve my skills since then) We have worked hard for over thirty years to begin to be recognized as professionals not just techs! DO NOT let this stop us now so the pathologists at ASCP and AMA can continue to treat Histology as minor part of the laboratory. Histology has changed and will continue to make strides forward that require more education not less. The board of NSH did send a letter to the ASCP and tell them they would not approve this category. It is a personal feeling however, I think the Board of NSH should have notified the membership about this immediately, as a letter is not sufficient to protect us or respond as members of both NSH and ASCP. Again, write ASCP/BOR about your feelings if this important to you. Second point Cathy Locallo put a motion forward at the House of Delegates to request a Task Force to see if there is way to get histologists (HT and HTL) working for the federal government recognized as professionals and here we have a new way with the HA registry to keep us status quo. By the time the task force is in place and moving we will be down a rung again. Pamela Marcum (This is a reflection of my opinion not my employer or any other person.) Pam Marcum _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From Eric <@t> ategra.com Fri Sep 16 14:34:29 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 21:43:17 2005 Subject: [Histonet] Immediate Job Opportunities For HistoTechs (perm and contract) Message-ID: Histonetters - I have temporary travel assignments and permanent HistoTech openings available for you right now throughout the Southeast and the rest of the country! If you are currently available, please contact me ASAP at (800) 466-9919, ext. 223. Our clients are now hiring and need to fill these positions ASAP. Thanks for your time/attention... Eric Dye (800) 466-9919, ext. 223 PS - If you know anyone else who is currently looking, I would appreciate it if you could email me there phone number/email address. (Your friend would also appreciate it too!) PSS - If YOU are interested in any of these opportunities, please call me ASAP @ (800) 466 9919 x223. To speed things up, please send me a copy of your resume, (if you haven't already done so) - There is absolutely N O C O S T to you whatsoever! Our services are entirely paid for by the hiring companies. Eric Dye (800) 466-9919, ext. 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792 (800)466-9919 ext 231 FAX: (407) 671-6075 eric@ategra.com To Learn More About Ategra: http://www.ategra.com ------------------------------------------------------------------------------- ------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again ------------------------------------------------------------------------------- ------- From petepath <@t> yahoo.com Sat Sep 17 10:08:22 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Sat Sep 17 10:08:38 2005 Subject: [Histonet] New ASCP Classification and You Message-ID: <20050917150822.45278.qmail@web30411.mail.mud.yahoo.com> I think this is a step backwards for this field. By approving of such a category they are sending the message this job is so easy you can just take anyone off the street give them a few lessons and out will pop your insitu hybridization. Your payscale will go backwards. I cannot imagine how anyone can understand what they are doing in this rather complex field without a basic knowledge of the scientific and technical principals you are using every day. Between the myriad of histochemical stains you need to know, you have now added immunohistochemistry and molecular techniques. These are very sophisticated scientific procedures. Think about the minutia of details and experience as well as technical information about your microtomes that is required just to get a nice clean unshattered ribbon of a GI biopsy. Sure, you can show someone the technical maneuvers in your free time but they will not reach nearly the same level of quality without a knowledge of the principals. As the guy reading the slides I would rather see your field require formal education and certification at all levels. I have no problem with OJ T's performing clerk duties, filing and such, but it is a bit scary to think of letting one of these loose on my biopsies or cover immunos becase every one is on vacation. My suggestion for these HA's is to designate an approved set of duties, much like doctors are given privileges for certain procedures and not others when we join a staff. Have them register with NSH as HA's in training. Make them complete classes and pass certification exams on a curriculum which is appropriate for their duties in a given time period. At least it would be like taking a correspondence course. Stephen From histomjans <@t> yahoo.com Sat Sep 17 13:52:06 2005 From: histomjans <@t> yahoo.com (Melissa Jans) Date: Sat Sep 17 13:52:28 2005 Subject: [Histonet] microwave for EM Message-ID: <20050917185206.4518.qmail@web54710.mail.yahoo.com> I hope this email finds everyone well, I have been charged with finding out if anyone is using a microwave to process EM specimens. If so, what kind of microwave are you using and what are the pros and cons of it? I would appreciate any feedback as we need to make a purchase soon and may not have the opportunity to demo. Thank you, Melissa Jans University of Iowa Hospitals and Clinics --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From pruegg <@t> ihctech.net Sun Sep 18 13:59:41 2005 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Sun Sep 18 14:00:27 2005 Subject: [Histonet] HT student Message-ID: <200509181859.j8IIxtjW076484@pro12.abac.com> I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net From lpwenk <@t> sbcglobal.net Sun Sep 18 17:37:57 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Sun Sep 18 17:38:56 2005 Subject: [Histonet] HT student In-Reply-To: <200509181859.j8IIxtjW076484@pro12.abac.com> Message-ID: <1085039814-324235194@pathology.swmed.edu> At our hospital lab, anyone we hire who is registry eligible has 1 1/2 years in which to take and pass their exam (HT/HTL/CT/QIHC, etc). It is stated/written when they are hired. If they do not take AND pass within that time period, they are let go. We made it for 1.5 years, as that gives them several attempts, in case they do fail. Everyone has taken and passed. Plus, they know that once they pass the exam, they get bumped up to the next pay level. Double incentive to take the exam early and pass it. As for the commitment, I know of some labs that have offered my students to pay sign-on bonuses, moving expenses, sometimes even out-right loans, with the understanding (in writing, of course) that these are "free loans", if the person comes to work for them for 1-2 (or 3) years. If the student leaves before the 1-2-3 years, they must repay the loans - either the entire amount, or the percentage left (in other words, if they agreed to work for 2 years (24 months), and they leave after 14 months, they had 10 months that they did not fulfill the agreement, so have to pay back 10/24 of the "loan". Just some thoughts. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pruegg@ihctech.net Sent: Sunday, September 18, 2005 3:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HT student I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sun Sep 18 17:43:32 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Sun Sep 18 17:44:23 2005 Subject: [Histonet] New ASCP Classification and You In-Reply-To: <432B0C4E.00000E.02344@YOUR-4DI1S53IME> Message-ID: Pam - I'm curious. Who at NSH is saying this? Renee Allegruci from the ASCP Board of Registry was at the NSH S/C (at the ASCP BOR booth in the exhibit hall). She attended the Instructors of Histotechnology meeting on Monday night. I asked her, in front of about a dozen histology schools' program directors, if ASCP BOR was pursuing the lab assistant exam route, and her answer was short - "No." Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pam marcum Sent: Friday, September 16, 2005 2:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You Good Morning, I have just returned from the NSH meeting. I was surprised to find out the Histology Assistant or HA category recently presented to the NSH board and refused with a letter is moving forward. No one had told the general membership of NSH or ASCP or had it been explained to them in full. It is important for those of us who have worked so hard to get the new the education rules set in place to understand what this new registry will be and how it will affect us as professionals. This is an aid position that is OJT (on the job training) and will mean the pathologist, hospital or other private histology laboratory will be able to avoid having more than registered HT or HTL in the laboratory. The fully registered HT or HTL can then be used as the supervisor or manager and responsible for all the HA's work. This will aid in maintaining the lower pay scale we currently enjoy and help prevent us raising our overall image as professionals. Please find out more about this and respond to the ASCP/BOR with comments about this new category. The only way any of us will be heard is to let ASCP know what we think and I hope many of you will join me in letting them know we do not want to go back to the 1960s and 70s when we were all OJT and the pay is still reflecting it. (By the way I was OJT and have worked hard to improve my skills since then) We have worked hard for over thirty years to begin to be recognized as professionals not just techs! DO NOT let this stop us now so the pathologists at ASCP and AMA can continue to treat Histology as minor part of the laboratory. Histology has changed and will continue to make strides forward that require more education not less. The board of NSH did send a letter to the ASCP and tell them they would not approve this category. It is a personal feeling however, I think the Board of NSH should have notified the membership about this immediately, as a letter is not sufficient to protect us or respond as members of both NSH and ASCP. Again, write ASCP/BOR about your feelings if this important to you. Second point Cathy Locallo put a motion forward at the House of Delegates to request a Task Force to see if there is way to get histologists (HT and HTL) working for the federal government recognized as professionals and here we have a new way with the HA registry to keep us status quo. By the time the task force is in place and moving we will be down a rung again. Pamela Marcum (This is a reflection of my opinion not my employer or any other person.) Pam Marcum _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From shieban <@t> gmail.com Mon Sep 19 00:15:27 2005 From: shieban <@t> gmail.com (Saeed Al-shieban) Date: Mon Sep 19 00:15:45 2005 Subject: [Histonet] training in muscle biopsies handling Message-ID: hi I am a medical tech. I am interested in handling of muscle biopsies. any body can advise me where is the best place to take training in that for 3 months at USA or UK thanks From HornHV <@t> archildrens.org Mon Sep 19 08:35:49 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Mon Sep 19 08:35:35 2005 Subject: [Histonet] HT student Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE21@EMAIL.archildrens.org> Just my thoughts but you should have required that of her before she started attending school with you footing part of the bill. I think it would be inappropriate to ask/demand it now. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Sunday, September 18, 2005 5:38 PM To: pruegg@ihctech.net; histonet@pathology.swmed.edu Subject: RE: [Histonet] HT student At our hospital lab, anyone we hire who is registry eligible has 1 1/2 years in which to take and pass their exam (HT/HTL/CT/QIHC, etc). It is stated/written when they are hired. If they do not take AND pass within that time period, they are let go. We made it for 1.5 years, as that gives them several attempts, in case they do fail. Everyone has taken and passed. Plus, they know that once they pass the exam, they get bumped up to the next pay level. Double incentive to take the exam early and pass it. As for the commitment, I know of some labs that have offered my students to pay sign-on bonuses, moving expenses, sometimes even out-right loans, with the understanding (in writing, of course) that these are "free loans", if the person comes to work for them for 1-2 (or 3) years. If the student leaves before the 1-2-3 years, they must repay the loans - either the entire amount, or the percentage left (in other words, if they agreed to work for 2 years (24 months), and they leave after 14 months, they had 10 months that they did not fulfill the agreement, so have to pay back 10/24 of the "loan". Just some thoughts. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pruegg@ihctech.net Sent: Sunday, September 18, 2005 3:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HT student I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From bliven.laura <@t> marshfieldclinic.org Mon Sep 19 09:13:53 2005 From: bliven.laura <@t> marshfieldclinic.org (bliven.laura@marshfieldclinic.org) Date: Mon Sep 19 09:14:19 2005 Subject: [Histonet] CAP Regulation Message-ID: <2904d01c5bd24$5d44a7e0$8e0110ac@mfldclinframe.org> The College of American Pathologists requires: "For immunohistochemistry tests that provide independent predictive/prognostic information, does the patient report include information on specimen processing, the antibody clone, and the scoring method used.?" Anyone willing to share their example? I'd like to keep it sweet, simple, and short, but have it look clean and easy to read. Please send by email or fax, void of all personal info please. Thanks, Laura Bliven Histology Lab/IHC Marshfield Laboratories 1000 N. Oak Ave. Marshfield, WI 54449 bliven.laura@marshfieldclinic.org fax#715-389-5353 From Jackie.O'Connor <@t> abbott.com Mon Sep 19 11:15:26 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Mon Sep 19 11:16:19 2005 Subject: [Histonet] Santa Cruz CD31 Message-ID: Does anyone know if Santa Cruz ever worked out the issue with their CD31 SC1506 antibody? Last I heard, their goat was sick. Is it safe to order this antibody with a reasonable expectation that it will work? Jackie O' From tahseen <@t> brain.net.pk Mon Sep 19 04:22:10 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Mon Sep 19 11:29:48 2005 Subject: [Histonet] OCT4 antibody for immunohistochemistry. Message-ID: <002401c5bcfb$9e1c5fc0$972bfea9@m7c0y4> Dear All, Our lab is looking for OCT4 antibody for immunohistochemistry identification of seminoma and embryonal carcinoma. Could you please provide us with the necessary information so that we can order the antibody. Thank you. Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. From la.sebree <@t> hosp.wisc.edu Mon Sep 19 11:34:33 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Mon Sep 19 11:34:55 2005 Subject: [Histonet] bcl-2 and bcl-6 for Ventana users Message-ID: Hello, I'm having some difficulty working up bcl-6 on our Ventana instruments and the bcl-2 we've always used isn't as robust as I'd like either. What clones and vendors are people using on their Ventana automated stainers? We have a NexES, a BenchMark and a BenchMark XT. Thanks for the info. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From pruegg <@t> ihctech.net Mon Sep 19 11:49:28 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Sep 19 11:49:51 2005 Subject: [Histonet] New ASCP Classification and You In-Reply-To: <20050917150822.45278.qmail@web30411.mail.mud.yahoo.com> Message-ID: <200509191649.j8JGnNfj002085@chip.viawest.net> Before we all get bent out of shape about this we need to verify that ASCP is really going forward with this classification or not. I am on the NSH BOD and at no time did we have any discussions about this with the assumption that ASCP is going forward with this. We need to ask Marilyn Gamble the NSH ASCP representative to clarify. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Saturday, September 17, 2005 8:08 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You I think this is a step backwards for this field. By approving of such a category they are sending the message this job is so easy you can just take anyone off the street give them a few lessons and out will pop your insitu hybridization. Your payscale will go backwards. I cannot imagine how anyone can understand what they are doing in this rather complex field without a basic knowledge of the scientific and technical principals you are using every day. Between the myriad of histochemical stains you need to know, you have now added immunohistochemistry and molecular techniques. These are very sophisticated scientific procedures. Think about the minutia of details and experience as well as technical information about your microtomes that is required just to get a nice clean unshattered ribbon of a GI biopsy. Sure, you can show someone the technical maneuvers in your free time but they will not reach nearly the same level of quality without a knowledge of the principals. As the guy reading the slides I would rather see your field require formal education and certification at all levels. I have no problem with OJ T's performing clerk duties, filing and such, but it is a bit scary to think of letting one of these loose on my biopsies or cover immunos becase every one is on vacation. My suggestion for these HA's is to designate an approved set of duties, much like doctors are given privileges for certain procedures and not others when we join a staff. Have them register with NSH as HA's in training. Make them complete classes and pass certification exams on a curriculum which is appropriate for their duties in a given time period. At least it would be like taking a correspondence course. Stephen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fredericoacazevedo <@t> gmail.com Mon Sep 19 12:42:24 2005 From: fredericoacazevedo <@t> gmail.com (frederico azevedo) Date: Mon Sep 19 12:42:39 2005 Subject: [Histonet] Antigen retrieval NeuN Message-ID: Hello everyone, Does anybody know the best antigen retrieval method to mark NeuN in human tissue? I?ve already tried some described by literature but I?ve got only weak imunostaining. I am using mouse monoclonal anti-Neun (Chemicon) as primary, an anti-mouse biotinilated secondary, ABC and VIP as chromogen. In rat, all neuronal nucleous are heavily stained therefore in human tissue... The brains I?m using are fixed in paraformaldehyde 4% for about 3 months and I try to mark 15 micrometers frozen sections... Is there any other neuronal nuclear marker for human tissue that is really specific? Thanks, Frederico A. C. Azevedo, Dept. of Anatomy - Neuroplasticity UFRJ, Brasil. From mucram11 <@t> comcast.net Mon Sep 19 12:50:05 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Mon Sep 19 12:50:46 2005 Subject: [Histonet] New ASCP Classification and You Message-ID: <091920051750.21719.432EFA4D0000231A000054D72200762194CECE030E9D0C9A03@comcast.net> Patsy, Since I am the one who originated the question and issue I will answer this also. My reasons for being upset were I had heard nothing about this until this year and I could not get an answer other than the board sent a letter to ASCP saying they would not condon the HA licsence last year. I ask a number of people and found the majority were completely unaware of the possible addition and had not been informed by either their region director, president, NSH board or NSH representative to ASCP or ASCP. This is something that can effect all of us still in the field and should have been disclosed and explained to the membership of NSH and Histologist in general by ASCP. As Marilyn Gamble is the reperesentative to ASCP perhaps she and the board should have let us know. If she asks ASCP now, I firmly believe this should be sent to everyone or published where it can be seen and understood as either a Yes or No. If ASCP and NSH are upset because of this question perhaps next time the disclosure will be more complete and timely. Pam Marcum -------------- Original message -------------- > Before we all get bent out of shape about this we need to verify that ASCP > is really going forward with this classification or not. I am on the NSH > BOD and at no time did we have any discussions about this with the > assumption that ASCP is going forward with this. We need to ask Marilyn > Gamble the NSH ASCP representative to clarify. > Patsy > > > Patsy Ruegg, HT(ASCP)QIHC > IHCtech, LLC > Fitzsimmons BioScience Park > 12635 Montview Blvd. Suite 216 > Aurora, CO 80010 > P-720-859-4060 > F-720-859-4110 > wk email pruegg@ihctech.net > web site www.ihctech.net > > > This email is confidential and intended solely for the use of the Person(s) > ('the intended recipient') to whom it was addressed. Any views or opinions > presented are solely those of the author. It may contain information that is > privileged & confidential within the meaning of applicable law. Accordingly > any dissemination, distribution, copying, or other use of this message, or > any of its contents, by any person other than the intended recipient may > constitute a breach of civil or criminal law and is strictly prohibited. If > you are NOT the intended recipient please contact the sender and dispose of > this e-mail as soon as possible. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen > Peters M.D. > Sent: Saturday, September 17, 2005 8:08 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] New ASCP Classification and You > > I think this is a step backwards for this field. By approving of such a > category they are sending the message this job is so easy you can just take > anyone off the street give them a few lessons and out will pop your insitu > hybridization. Your payscale will go backwards. > I cannot imagine how anyone can understand what they are doing in this > rather complex field without a basic knowledge of the scientific and > technical principals you are using every day. Between the myriad of > histochemical stains you need to know, you have now added > immunohistochemistry and molecular techniques. These are very sophisticated > scientific procedures. Think about the minutia of details and experience as > well as technical information about your microtomes that is required just to > get a nice clean unshattered ribbon of a GI biopsy. Sure, you can show > someone the technical maneuvers in your free time but they will not reach > nearly the same level of quality without a knowledge of the principals. As > the guy reading the slides I would rather see your field require formal > education and certification at all levels. I have no problem with OJ T's > performing clerk duties, filing and such, but it is a bit scary to think of > letting one of these loose on my biopsies or cover immunos becase every one > is on vacation. > > My suggestion for these HA's is to designate an approved set of duties, much > like doctors are given privileges for certain procedures and not others when > we join a staff. Have them register with NSH as HA's in training. Make them > complete classes and pass certification exams on a curriculum which is > appropriate for their duties in a given time period. At least it would be > like taking a correspondence course. > > Stephen > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Cathy.Crumpton <@t> tuality.org Mon Sep 19 12:51:22 2005 From: Cathy.Crumpton <@t> tuality.org (Cathy.Crumpton@tuality.org) Date: Mon Sep 19 12:51:51 2005 Subject: [Histonet] larger pre-filled formalin containers Message-ID: Does anyone know of a company in the US that carries pre-filled f ormalin containers in the 1L and 2L size? I found a place in Canada t Cathy < From Melissa.Gonzalez <@t> cellgenesys.com Mon Sep 19 13:00:32 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Mon Sep 19 13:01:34 2005 Subject: [Histonet] Santa Cruz Pecam-1 Message-ID: Hi Jackie, I recently tried this product again and it still doesn't work. I suggest using CD105 or CD34, in the meantime. I haven't found a suitable CD31 replacement to compare to how great Santa Cruz's product used to work. Melissa Melissa A. Gonzalez Histology Cell Genesys 500 Forbes Blvd. South San Francisco, CA 94080 -----Original Message----- Message: 7 Date: Mon, 19 Sep 2005 11:15:26 -0500 From: "Jackie M O'Connor" Subject: [Histonet] Santa Cruz CD31 To: histonet@pathology.swmed.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Does anyone know if Santa Cruz ever worked out the issue with their CD31 SC1506 antibody? Last I heard, their goat was sick. Is it safe to order this antibody with a reasonable expectation that it will work? Jackie O' From POWELL_SA <@t> Mercer.edu Mon Sep 19 13:31:15 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Mon Sep 19 13:32:28 2005 Subject: [Histonet] larger pre-filled formalin containers In-Reply-To: Message-ID: <01LT7VRP5ABI8X0B15@Macon2.Mercer.edu> Check with Polyscientific. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cathy.Crumpton@tuality.org Sent: Monday, September 19, 2005 12:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] larger pre-filled formalin containers Does anyone know of a company in the US that carries pre-filled f ormalin containers in the 1L and 2L size? I found a place in Canada t hat does but don't want to pay extra shipping. Cathy < /FONT>_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Mon Sep 19 15:37:20 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Sep 19 15:37:42 2005 Subject: [Histonet] Anti-human Vegf Message-ID: <200509192037.j8JKbFfj012254@chip.viawest.net> OK, I got monoclonal vegf from Santa Cruz to work on human samples a few years ago, but now I am trying to use it on Baboon and it is not right. Is it the inconsistency in SC antibodies that is the problem? Does anyone know of any other monoclonal anti-human vegf? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From sbreeden <@t> nmda.nmsu.edu Mon Sep 19 16:02:57 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Mon Sep 19 16:03:16 2005 Subject: [Histonet] Sakura-esque slide racks Message-ID: I'm on the hunt for those Sakura 20-slide racks for tape coverslippers; handles, too. I'm sure this has been covered on 'net before, but could someone direct me? Vendors are welcome to contact me, but be it known that I'm only collecting information at this point... Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From pruegg <@t> ihctech.net Mon Sep 19 16:26:08 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Sep 19 16:26:51 2005 Subject: [Histonet] New ASCP Preparatory Tech Classification Message-ID: <200509192126.j8JLQ8fj026599@chip.viawest.net> I have just gotten back confirmation from Marilyn Gamble (the NSH representative to ASCP) as follows: "The Prepatory Tech classification was not approved by the ASCP Board of Governors. It was dropped." Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From mary.ann.deathridge <@t> Vanderbilt.Edu Mon Sep 19 16:30:00 2005 From: mary.ann.deathridge <@t> Vanderbilt.Edu (Deathridge, Mary Ann) Date: Mon Sep 19 16:30:24 2005 Subject: [Histonet] digest Message-ID: <59342A85CAA485429AF89AAA42FFD9DF36B73E@mailbe10.mc.vanderbilt.edu> Thanks MaryAnn Deathridge, HT (ASCP) Supervisor, Histopathology TVC 4532 3-7012 From jnocito <@t> satx.rr.com Mon Sep 19 18:47:22 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Mon Sep 19 18:45:11 2005 Subject: [Histonet] New ASCP Preparatory Tech Classification References: <200509192126.j8JLQ8fj026599@chip.viawest.net> Message-ID: <004d01c5bd74$7b1002b0$52bf0b43@yourxhtr8hvc4p> it's about time ASCP did something good for histology. Still haven't gotten over the $300 increase in taking the PA exam. Shucks, just as I was going to jump on the band wagon with Pam. These last couple of weeks just hasn't been fun for me. Give me a while, I'm sure there is something out there I can rustle my feathers over. Oh yeah, I must be feeling better. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Patsy Ruegg" To: "'Histonet'" Sent: Monday, September 19, 2005 4:26 PM Subject: [Histonet] New ASCP Preparatory Tech Classification >I have just gotten back confirmation from Marilyn Gamble (the NSH > representative to ASCP) as follows: > "The Prepatory Tech classification was not approved by the ASCP Board of > Governors. It was dropped." > Patsy > > > Patsy Ruegg, HT(ASCP)QIHC > IHCtech, LLC > Fitzsimmons BioScience Park > 12635 Montview Blvd. Suite 216 > Aurora, CO 80010 > P-720-859-4060 > F-720-859-4110 > wk email pruegg@ihctech.net > web site www.ihctech.net > > > This email is confidential and intended solely for the use of the > Person(s) > ('the intended recipient') to whom it was addressed. Any views or opinions > presented are solely those of the author. It may contain information that > is > privileged & confidential within the meaning of applicable law. > Accordingly > any dissemination, distribution, copying, or other use of this message, or > any of its contents, by any person other than the intended recipient may > constitute a breach of civil or criminal law and is strictly prohibited. > If > you are NOT the intended recipient please contact the sender and dispose > of > this e-mail as soon as possible. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.344 / Virus Database: 267.11.2/105 - Release Date: 9/19/2005 > > From Jason.PALMER <@t> svhm.org.au Mon Sep 19 20:16:40 2005 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Mon Sep 19 20:17:21 2005 Subject: [Histonet] Santa Cruz CD31 SC1506 Message-ID: Jackie, I queried Santa Cruz re this antibody in May this year, and below I have pasted part of their response to me - to summarize, a new batch is expected "by the end of September", which is just about upon us, obviously. Cheers, Jason Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 fax +61 3 9416 0926 email: palmerj@svhm.org.au ----- Message: 7 Date: Mon, 19 Sep 2005 11:15:26 -0500 From: "Jackie M O'Connor" Subject: [Histonet] Santa Cruz CD31 To: histonet@pathology.swmed.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Does anyone know if Santa Cruz ever worked out the issue with their CD31 SC1506 antibody? Last I heard, their goat was sick. Is it safe to order this antibody with a reasonable expectation that it will work? Jackie O' ---Original Message----- From: Julia Renstrom [mailto:renstromj@scbt.com] Sent: Fri 5/20/2005 3:34 AM To: PALMER Jason (SVHM) Cc: Subject: Re: problems with Santa Cruz sc-1506 antibody Dear Dr. Palmer, Thank you for your inquiry. This antibody, sc-1506, is usually one of the strongest PECAM1 antibodies in the industry. However, we have seen some inconsistent staining of paraffin-embedded, formalin-fixed sections in recent lots of this antibody. However we had some problems with the goat that used to produce the IgG and immunized a new goat with the sc-1506 peptide immunogen. The new goat which is currently host to this antibody is producing IgG which is still performing in Western blot, but for some reason is not staining at the level we expect for paraffin-embedded sections. Unfortunately, this is also the only appropriate product we can offer for your purposes -- it is the only mouse-reactive antibody which we recommend for paraffin-embedded sections. We have immunized yet another goat with the sc-1506 peptide immunogen and predict a release of this new IgG by the end of September, pending completion of our quality control testing. -- Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From vanann702 <@t> skmc.gov.ae Mon Sep 19 22:52:27 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Mon Sep 19 22:52:32 2005 Subject: [Histonet] Sakura-esque slide racks Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CBFF@SKMCEMAIL.skmc.gov.ae> Try the Sakura website - they have them Annieinarabia -----Original Message----- From: Breeden, Sara [mailto:sbreeden@nmda.nmsu.edu] Sent: Tuesday, September 20, 2005 1:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sakura-esque slide racks I'm on the hunt for those Sakura 20-slide racks for tape coverslippers; handles, too. I'm sure this has been covered on 'net before, but could someone direct me? Vendors are welcome to contact me, but be it known that I'm only collecting information at this point... Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brucea <@t> unimelb.edu.au Tue Sep 20 01:48:03 2005 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Tue Sep 20 01:48:53 2005 Subject: [Histonet] Marine Blood Smears Message-ID: Hi, I am a Vet doing Research in Zoology RE: abalone blood cells. If I air dry my fresh blood smears I get terrible salt crystal formation which interferes with reading the smears. Any ideas on how to fix/stain my blood smears to get rid of this problem? I am sure the folks who do work with marine species of fish would have encountered the same problem in fresh blood smears. I want to do the smears to do differential counts, examine morphology etc, same as for any other species. Thanks ahead for any advice. -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA.AUSTRALIA 3010 http://www.zoology.unimelb.edu.au/ Nobody can make you feel inferior without your permission. - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue Sep 20 02:38:32 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Tue Sep 20 02:41:00 2005 Subject: [Histonet] RE: Bcl 2 and Benchmark XT Message-ID: We use Dako's clone 124 (code M 0887) and the protocol mild CC1, incubate at 37 for 1 hour with amplification. It was one of the last to be optimised and isn't as strong as when we used the manual method. Hope this helps Jacqui Malam Royal Lancaster Infirmary Lancaster N. England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From AFeatherstone <@t> KaleidaHealth.Org Tue Sep 20 05:26:03 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Tue Sep 20 05:24:50 2005 Subject: [Histonet] RE: Histonet Digest, Vol 22, Issue 22 Message-ID: <139141F8BAF4A642A945ECC528511AF0012A7939@kalmb02.kaleidahealth.org> I would like to know how everyone is disposing of the ammonium-silver nitrate waste and DAB. Any and all responses would be of great help. Thanks Annette Featherstone HT/MLT Supervisor Anatomic Pathology Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 716-859-2625 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, September 19, 2005 13:01 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 22, Issue 22 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. HT student (pruegg@ihctech.net) 2. RE: HT student (Lee & Peggy Wenk) 3. RE: New ASCP Classification and You (Lee & Peggy Wenk) 4. training in muscle biopsies handling (Saeed Al-shieban) 5. RE: HT student (Horn, Hazel V) 6. CAP Regulation (bliven.laura@marshfieldclinic.org) 7. Santa Cruz CD31 (Jackie M O'Connor) 8. OCT4 antibody for immunohistochemistry. (Muhammad Tahseen) 9. bcl-2 and bcl-6 for Ventana users (Sebree Linda A.) 10. RE: New ASCP Classification and You (Patsy Ruegg) ---------------------------------------------------------------------- Message: 1 Date: Sun, 18 Sep 2005 12:59:41 -0600 From: pruegg@ihctech.net Subject: [Histonet] HT student To: Message-ID: <200509181859.j8IIxtjW076484@pro12.abac.com> Content-Type: text/plain; charset="us-ascii" I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net ------------------------------ Message: 2 Date: Sun, 18 Sep 2005 18:37:57 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] HT student To: , Message-ID: <1085039814-324235194@pathology.swmed.edu> Content-Type: text/plain; charset="us-ascii" At our hospital lab, anyone we hire who is registry eligible has 1 1/2 years in which to take and pass their exam (HT/HTL/CT/QIHC, etc). It is stated/written when they are hired. If they do not take AND pass within that time period, they are let go. We made it for 1.5 years, as that gives them several attempts, in case they do fail. Everyone has taken and passed. Plus, they know that once they pass the exam, they get bumped up to the next pay level. Double incentive to take the exam early and pass it. As for the commitment, I know of some labs that have offered my students to pay sign-on bonuses, moving expenses, sometimes even out-right loans, with the understanding (in writing, of course) that these are "free loans", if the person comes to work for them for 1-2 (or 3) years. If the student leaves before the 1-2-3 years, they must repay the loans - either the entire amount, or the percentage left (in other words, if they agreed to work for 2 years (24 months), and they leave after 14 months, they had 10 months that they did not fulfill the agreement, so have to pay back 10/24 of the "loan". Just some thoughts. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pruegg@ihctech.net Sent: Sunday, September 18, 2005 3:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HT student I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Sun, 18 Sep 2005 18:43:32 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] New ASCP Classification and You To: "'pam marcum'" , Message-ID: Content-Type: text/plain; charset="us-ascii" Pam - I'm curious. Who at NSH is saying this? Renee Allegruci from the ASCP Board of Registry was at the NSH S/C (at the ASCP BOR booth in the exhibit hall). She attended the Instructors of Histotechnology meeting on Monday night. I asked her, in front of about a dozen histology schools' program directors, if ASCP BOR was pursuing the lab assistant exam route, and her answer was short - "No." Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pam marcum Sent: Friday, September 16, 2005 2:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You Good Morning, I have just returned from the NSH meeting. I was surprised to find out the Histology Assistant or HA category recently presented to the NSH board and refused with a letter is moving forward. No one had told the general membership of NSH or ASCP or had it been explained to them in full. It is important for those of us who have worked so hard to get the new the education rules set in place to understand what this new registry will be and how it will affect us as professionals. This is an aid position that is OJT (on the job training) and will mean the pathologist, hospital or other private histology laboratory will be able to avoid having more than registered HT or HTL in the laboratory. The fully registered HT or HTL can then be used as the supervisor or manager and responsible for all the HA's work. This will aid in maintaining the lower pay scale we currently enjoy and help prevent us raising our overall image as professionals. Please find out more about this and respond to the ASCP/BOR with comments about this new category. The only way any of us will be heard is to let ASCP know what we think and I hope many of you will join me in letting them know we do not want to go back to the 1960s and 70s when we were all OJT and the pay is still reflecting it. (By the way I was OJT and have worked hard to improve my skills since then) We have worked hard for over thirty years to begin to be recognized as professionals not just techs! DO NOT let this stop us now so the pathologists at ASCP and AMA can continue to treat Histology as minor part of the laboratory. Histology has changed and will continue to make strides forward that require more education not less. The board of NSH did send a letter to the ASCP and tell them they would not approve this category. It is a personal feeling however, I think the Board of NSH should have notified the membership about this immediately, as a letter is not sufficient to protect us or respond as members of both NSH and ASCP. Again, write ASCP/BOR about your feelings if this important to you. Second point Cathy Locallo put a motion forward at the House of Delegates to request a Task Force to see if there is way to get histologists (HT and HTL) working for the federal government recognized as professionals and here we have a new way with the HA registry to keep us status quo. By the time the task force is in place and moving we will be down a rung again. Pamela Marcum (This is a reflection of my opinion not my employer or any other person.) Pam Marcum _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 19 Sep 2005 08:15:27 +0300 From: Saeed Al-shieban Subject: [Histonet] training in muscle biopsies handling To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 hi I am a medical tech. I am interested in handling of muscle biopsies. any body can advise me where is the best place to take training in that for 3 months at USA or UK thanks ------------------------------ Message: 5 Date: Mon, 19 Sep 2005 08:35:49 -0500 From: "Horn, Hazel V" Subject: RE: [Histonet] HT student To: pruegg@ihctech.net, Histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE21@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii Just my thoughts but you should have required that of her before she started attending school with you footing part of the bill. I think it would be inappropriate to ask/demand it now. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Sunday, September 18, 2005 5:38 PM To: pruegg@ihctech.net; histonet@pathology.swmed.edu Subject: RE: [Histonet] HT student At our hospital lab, anyone we hire who is registry eligible has 1 1/2 years in which to take and pass their exam (HT/HTL/CT/QIHC, etc). It is stated/written when they are hired. If they do not take AND pass within that time period, they are let go. We made it for 1.5 years, as that gives them several attempts, in case they do fail. Everyone has taken and passed. Plus, they know that once they pass the exam, they get bumped up to the next pay level. Double incentive to take the exam early and pass it. As for the commitment, I know of some labs that have offered my students to pay sign-on bonuses, moving expenses, sometimes even out-right loans, with the understanding (in writing, of course) that these are "free loans", if the person comes to work for them for 1-2 (or 3) years. If the student leaves before the 1-2-3 years, they must repay the loans - either the entire amount, or the percentage left (in other words, if they agreed to work for 2 years (24 months), and they leave after 14 months, they had 10 months that they did not fulfill the agreement, so have to pay back 10/24 of the "loan". Just some thoughts. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pruegg@ihctech.net Sent: Sunday, September 18, 2005 3:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HT student I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == ------------------------------ Message: 6 Date: Mon, 19 Sep 2005 09:13:53 -0500 From: bliven.laura@marshfieldclinic.org Subject: [Histonet] CAP Regulation To: Message-ID: <2904d01c5bd24$5d44a7e0$8e0110ac@mfldclinframe.org> The College of American Pathologists requires: "For immunohistochemistry tests that provide independent predictive/prognostic information, does the patient report include information on specimen processing, the antibody clone, and the scoring method used.?" Anyone willing to share their example? I'd like to keep it sweet, simple, and short, but have it look clean and easy to read. Please send by email or fax, void of all personal info please. Thanks, Laura Bliven Histology Lab/IHC Marshfield Laboratories 1000 N. Oak Ave. Marshfield, WI 54449 bliven.laura@marshfieldclinic.org fax#715-389-5353 ------------------------------ Message: 7 Date: Mon, 19 Sep 2005 11:15:26 -0500 From: "Jackie M O'Connor" Subject: [Histonet] Santa Cruz CD31 To: histonet@pathology.swmed.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Does anyone know if Santa Cruz ever worked out the issue with their CD31 SC1506 antibody? Last I heard, their goat was sick. Is it safe to order this antibody with a reasonable expectation that it will work? Jackie O' ------------------------------ Message: 8 Date: Mon, 19 Sep 2005 21:22:10 +1200 From: "Muhammad Tahseen" Subject: [Histonet] OCT4 antibody for immunohistochemistry. To: "HistoNet Server" Message-ID: <002401c5bcfb$9e1c5fc0$972bfea9@m7c0y4> Content-Type: text/plain; charset="iso-8859-1" Dear All, Our lab is looking for OCT4 antibody for immunohistochemistry identification of seminoma and embryonal carcinoma. Could you please provide us with the necessary information so that we can order the antibody. Thank you. Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. ------------------------------ Message: 9 Date: Mon, 19 Sep 2005 11:34:33 -0500 From: "Sebree Linda A." Subject: [Histonet] bcl-2 and bcl-6 for Ventana users To: "Histonet \(Histonet\)" Message-ID: Content-Type: text/plain; charset="us-ascii" Hello, I'm having some difficulty working up bcl-6 on our Ventana instruments and the bcl-2 we've always used isn't as robust as I'd like either. What clones and vendors are people using on their Ventana automated stainers? We have a NexES, a BenchMark and a BenchMark XT. Thanks for the info. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 ------------------------------ Message: 10 Date: Mon, 19 Sep 2005 10:49:28 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] New ASCP Classification and You To: "'Stephen Peters M.D.'" , Message-ID: <200509191649.j8JGnNfj002085@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Before we all get bent out of shape about this we need to verify that ASCP is really going forward with this classification or not. I am on the NSH BOD and at no time did we have any discussions about this with the assumption that ASCP is going forward with this. We need to ask Marilyn Gamble the NSH ASCP representative to clarify. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Saturday, September 17, 2005 8:08 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You I think this is a step backwards for this field. By approving of such a category they are sending the message this job is so easy you can just take anyone off the street give them a few lessons and out will pop your insitu hybridization. Your payscale will go backwards. I cannot imagine how anyone can understand what they are doing in this rather complex field without a basic knowledge of the scientific and technical principals you are using every day. Between the myriad of histochemical stains you need to know, you have now added immunohistochemistry and molecular techniques. These are very sophisticated scientific procedures. Think about the minutia of details and experience as well as technical information about your microtomes that is required just to get a nice clean unshattered ribbon of a GI biopsy. Sure, you can show someone the technical maneuvers in your free time but they will not reach nearly the same level of quality without a knowledge of the principals. As the guy reading the slides I would rather see your field require formal education and certification at all levels. I have no problem with OJ T's performing clerk duties, filing and such, but it is a bit scary to think of letting one of these loose on my biopsies or cover immunos becase every one is on vacation. My suggestion for these HA's is to designate an approved set of duties, much like doctors are given privileges for certain procedures and not others when we join a staff. Have them register with NSH as HA's in training. Make them complete classes and pass certification exams on a curriculum which is appropriate for their duties in a given time period. At least it would be like taking a correspondence course. Stephen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 22 **************************************** CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From rjbuesa <@t> yahoo.com Tue Sep 20 07:16:25 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Sep 20 07:16:48 2005 Subject: [Histonet] RE: Histonet Digest, Vol 22, Issue 22: DAB disposal. In-Reply-To: <139141F8BAF4A642A945ECC528511AF0012A7939@kalmb02.kaleidahealth.org> Message-ID: <20050920121625.34917.qmail@web61214.mail.yahoo.com> Annette: The practice of treating DAB with bleach is not recommended as the final product is unkown. According with Lunn & Sansone (1990) DAB can be destroyed as follows: 1- prepare a 0.2 M aqueous solution of potassium permanganate (31.6 g/litre). 2- prepare a2.0 M solution of sulfuric acid (112 ml of conc. acid/litre). 3- dilute the DAB in a way that its final concentration does not excede 0.9 mg/ml; you will have to do these calculations according with the DAB solution you use). 4 To EACH 10 ml of DAB solution add 5 ml of solution 1 and 5 ml of solution 2. 5- Allow the mixture for at least 10 hours. Now the solution will be NON mutagenic. 6- Add ascorbic acid (powder) until all color disappears). 7- Neutralize solution 6 with sodium bicarbonate (test with pH paper to pH=7). 8- Discard the solution down the drain PROVIDED that local water authorities have given their approval (which is something much more difficult that following the above procedure). Silver nitrata is a "horse of a different color" being a metal (not very heavy, but a metal nevertheless). I advise you to talk with the personnel in Radiology because they deal with this product and recover the silver. Rene J. Buesa "Featherstone, Annette" wrote: I would like to know how everyone is disposing of the ammonium-silver nitrate waste and DAB. Any and all responses would be of great help. Thanks Annette Featherstone HT/MLT Supervisor Anatomic Pathology Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 716-859-2625 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, September 19, 2005 13:01 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 22, Issue 22 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. HT student (pruegg@ihctech.net) 2. RE: HT student (Lee & Peggy Wenk) 3. RE: New ASCP Classification and You (Lee & Peggy Wenk) 4. training in muscle biopsies handling (Saeed Al-shieban) 5. RE: HT student (Horn, Hazel V) 6. CAP Regulation (bliven.laura@marshfieldclinic.org) 7. Santa Cruz CD31 (Jackie M O'Connor) 8. OCT4 antibody for immunohistochemistry. (Muhammad Tahseen) 9. bcl-2 and bcl-6 for Ventana users (Sebree Linda A.) 10. RE: New ASCP Classification and You (Patsy Ruegg) ---------------------------------------------------------------------- Message: 1 Date: Sun, 18 Sep 2005 12:59:41 -0600 From: pruegg@ihctech.net Subject: [Histonet] HT student To: Message-ID: <200509181859.j8IIxtjW076484@pro12.abac.com> Content-Type: text/plain; charset="us-ascii" I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net ------------------------------ Message: 2 Date: Sun, 18 Sep 2005 18:37:57 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] HT student To: , Message-ID: <1085039814-324235194@pathology.swmed.edu> Content-Type: text/plain; charset="us-ascii" At our hospital lab, anyone we hire who is registry eligible has 1 1/2 years in which to take and pass their exam (HT/HTL/CT/QIHC, etc). It is stated/written when they are hired. If they do not take AND pass within that time period, they are let go. We made it for 1.5 years, as that gives them several attempts, in case they do fail. Everyone has taken and passed. Plus, they know that once they pass the exam, they get bumped up to the next pay level. Double incentive to take the exam early and pass it. As for the commitment, I know of some labs that have offered my students to pay sign-on bonuses, moving expenses, sometimes even out-right loans, with the understanding (in writing, of course) that these are "free loans", if the person comes to work for them for 1-2 (or 3) years. If the student leaves before the 1-2-3 years, they must repay the loans - either the entire amount, or the percentage left (in other words, if they agreed to work for 2 years (24 months), and they leave after 14 months, they had 10 months that they did not fulfill the agreement, so have to pay back 10/24 of the "loan". Just some thoughts. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pruegg@ihctech.net Sent: Sunday, September 18, 2005 3:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HT student I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Sun, 18 Sep 2005 18:43:32 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] New ASCP Classification and You To: "'pam marcum'" , Message-ID: Content-Type: text/plain; charset="us-ascii" Pam - I'm curious. Who at NSH is saying this? Renee Allegruci from the ASCP Board of Registry was at the NSH S/C (at the ASCP BOR booth in the exhibit hall). She attended the Instructors of Histotechnology meeting on Monday night. I asked her, in front of about a dozen histology schools' program directors, if ASCP BOR was pursuing the lab assistant exam route, and her answer was short - "No." Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pam marcum Sent: Friday, September 16, 2005 2:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You Good Morning, I have just returned from the NSH meeting. I was surprised to find out the Histology Assistant or HA category recently presented to the NSH board and refused with a letter is moving forward. No one had told the general membership of NSH or ASCP or had it been explained to them in full. It is important for those of us who have worked so hard to get the new the education rules set in place to understand what this new registry will be and how it will affect us as professionals. This is an aid position that is OJT (on the job training) and will mean the pathologist, hospital or other private histology laboratory will be able to avoid having more than registered HT or HTL in the laboratory. The fully registered HT or HTL can then be used as the supervisor or manager and responsible for all the HA's work. This will aid in maintaining the lower pay scale we currently enjoy and help prevent us raising our overall image as professionals. Please find out more about this and respond to the ASCP/BOR with comments about this new category. The only way any of us will be heard is to let ASCP know what we think and I hope many of you will join me in letting them know we do not want to go back to the 1960s and 70s when we were all OJT and the pay is still reflecting it. (By the way I was OJT and have worked hard to improve my skills since then) We have worked hard for over thirty years to begin to be recognized as professionals not just techs! DO NOT let this stop us now so the pathologists at ASCP and AMA can continue to treat Histology as minor part of the laboratory. Histology has changed and will continue to make strides forward that require more education not less. The board of NSH did send a letter to the ASCP and tell them they would not approve this category. It is a personal feeling however, I think the Board of NSH should have notified the membership about this immediately, as a letter is not sufficient to protect us or respond as members of both NSH and ASCP. Again, write ASCP/BOR about your feelings if this important to you. Second point Cathy Locallo put a motion forward at the House of Delegates to request a Task Force to see if there is way to get histologists (HT and HTL) working for the federal government recognized as professionals and here we have a new way with the HA registry to keep us status quo. By the time the task force is in place and moving we will be down a rung again. Pamela Marcum (This is a reflection of my opinion not my employer or any other person.) Pam Marcum _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 19 Sep 2005 08:15:27 +0300 From: Saeed Al-shieban Subject: [Histonet] training in muscle biopsies handling To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 hi I am a medical tech. I am interested in handling of muscle biopsies. any body can advise me where is the best place to take training in that for 3 months at USA or UK thanks ------------------------------ Message: 5 Date: Mon, 19 Sep 2005 08:35:49 -0500 From: "Horn, Hazel V" Subject: RE: [Histonet] HT student To: pruegg@ihctech.net, Histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE21@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii Just my thoughts but you should have required that of her before she started attending school with you footing part of the bill. I think it would be inappropriate to ask/demand it now. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Sunday, September 18, 2005 5:38 PM To: pruegg@ihctech.net; histonet@pathology.swmed.edu Subject: RE: [Histonet] HT student At our hospital lab, anyone we hire who is registry eligible has 1 1/2 years in which to take and pass their exam (HT/HTL/CT/QIHC, etc). It is stated/written when they are hired. If they do not take AND pass within that time period, they are let go. We made it for 1.5 years, as that gives them several attempts, in case they do fail. Everyone has taken and passed. Plus, they know that once they pass the exam, they get bumped up to the next pay level. Double incentive to take the exam early and pass it. As for the commitment, I know of some labs that have offered my students to pay sign-on bonuses, moving expenses, sometimes even out-right loans, with the understanding (in writing, of course) that these are "free loans", if the person comes to work for them for 1-2 (or 3) years. If the student leaves before the 1-2-3 years, they must repay the loans - either the entire amount, or the percentage left (in other words, if they agreed to work for 2 years (24 months), and they leave after 14 months, they had 10 months that they did not fulfill the agreement, so have to pay back 10/24 of the "loan". Just some thoughts. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pruegg@ihctech.net Sent: Sunday, September 18, 2005 3:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HT student I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================ == ------------------------------ Message: 6 Date: Mon, 19 Sep 2005 09:13:53 -0500 From: bliven.laura@marshfieldclinic.org Subject: [Histonet] CAP Regulation To: Message-ID: <2904d01c5bd24$5d44a7e0$8e0110ac@mfldclinframe.org> The College of American Pathologists requires: "For immunohistochemistry tests that provide independent predictive/prognostic information, does the patient report include information on specimen processing, the antibody clone, and the scoring method used.?" Anyone willing to share their example? I'd like to keep it sweet, simple, and short, but have it look clean and easy to read. Please send by email or fax, void of all personal info please. Thanks, Laura Bliven Histology Lab/IHC Marshfield Laboratories 1000 N. Oak Ave. Marshfield, WI 54449 bliven.laura@marshfieldclinic.org fax#715-389-5353 ------------------------------ Message: 7 Date: Mon, 19 Sep 2005 11:15:26 -0500 From: "Jackie M O'Connor" Subject: [Histonet] Santa Cruz CD31 To: histonet@pathology.swmed.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Does anyone know if Santa Cruz ever worked out the issue with their CD31 SC1506 antibody? Last I heard, their goat was sick. Is it safe to order this antibody with a reasonable expectation that it will work? Jackie O' ------------------------------ Message: 8 Date: Mon, 19 Sep 2005 21:22:10 +1200 From: "Muhammad Tahseen" Subject: [Histonet] OCT4 antibody for immunohistochemistry. To: "HistoNet Server" Message-ID: <002401c5bcfb$9e1c5fc0$972bfea9@m7c0y4> Content-Type: text/plain; charset="iso-8859-1" Dear All, Our lab is looking for OCT4 antibody for immunohistochemistry identification of seminoma and embryonal carcinoma. Could you please provide us with the necessary information so that we can order the antibody. Thank you. Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. ------------------------------ Message: 9 Date: Mon, 19 Sep 2005 11:34:33 -0500 From: "Sebree Linda A." Subject: [Histonet] bcl-2 and bcl-6 for Ventana users To: "Histonet \(Histonet\)" Message-ID: Content-Type: text/plain; charset="us-ascii" Hello, I'm having some difficulty working up bcl-6 on our Ventana instruments and the bcl-2 we've always used isn't as robust as I'd like either. What clones and vendors are people using on their Ventana automated stainers? We have a NexES, a BenchMark and a BenchMark XT. Thanks for the info. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 ------------------------------ Message: 10 Date: Mon, 19 Sep 2005 10:49:28 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] New ASCP Classification and You To: "'Stephen Peters M.D.'" , Message-ID: <200509191649.j8JGnNfj002085@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Before we all get bent out of shape about this we need to verify that ASCP is really going forward with this classification or not. I am on the NSH BOD and at no time did we have any discussions about this with the assumption that ASCP is going forward with this. We need to ask Marilyn Gamble the NSH ASCP representative to clarify. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Saturday, September 17, 2005 8:08 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You I think this is a step backwards for this field. By approving of such a category they are sending the message this job is so easy you can just take anyone off the street give them a few lessons and out will pop your insitu hybridization. Your payscale will go backwards. I cannot imagine how anyone can understand what they are doing in this rather complex field without a basic knowledge of the scientific and technical principals you are using every day. Between the myriad of histochemical stains you need to know, you have now added immunohistochemistry and molecular techniques. These are very sophisticated scientific procedures. Think about the minutia of details and experience as well as technical information about your microtomes that is required just to get a nice clean unshattered ribbon of a GI biopsy. Sure, you can show someone the technical maneuvers in your free time but they will not reach nearly the same level of quality without a knowledge of the principals. As the guy reading the slides I would rather see your field require formal education and certification at all levels. I have no problem with OJ T's performing clerk duties, filing and such, but it is a bit scary to think of letting one of these loose on my biopsies or cover immunos becase every one is on vacation. My suggestion for these HA's is to designate an approved set of duties, much like doctors are given privileges for certain procedures and not others when === message truncated === --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From salder <@t> ventanamed.com Tue Sep 20 08:02:03 2005 From: salder <@t> ventanamed.com (Sandra Alder) Date: Tue Sep 20 08:02:32 2005 Subject: [Histonet] bcl-2 and bcl-6 clones Message-ID: <82322D77A7CFF04B888D2E9344C44A4402BF789A@GRIEVOUS.ventana.ventanamed.com> I have a customer who has good success with these, and I will ask her what she is using. Ventana has a new bcl-2 (clone 124) predilute, and a bcl-6 that is coming soon that you may also want to try. Sandra Alder Account Executive Ventana Medical Systems, Inc. 1 800 227 2155 ext. 2819 salder@ventanamed.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, September 19, 2005 1:03 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 22, Issue 22 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. HT student (pruegg@ihctech.net) 2. RE: HT student (Lee & Peggy Wenk) 3. RE: New ASCP Classification and You (Lee & Peggy Wenk) 4. training in muscle biopsies handling (Saeed Al-shieban) 5. RE: HT student (Horn, Hazel V) 6. CAP Regulation (bliven.laura@marshfieldclinic.org) 7. Santa Cruz CD31 (Jackie M O'Connor) 8. OCT4 antibody for immunohistochemistry. (Muhammad Tahseen) 9. bcl-2 and bcl-6 for Ventana users (Sebree Linda A.) 10. RE: New ASCP Classification and You (Patsy Ruegg) ---------------------------------------------------------------------- Message: 1 Date: Sun, 18 Sep 2005 12:59:41 -0600 From: pruegg@ihctech.net Subject: [Histonet] HT student To: Message-ID: <200509181859.j8IIxtjW076484@pro12.abac.com> Content-Type: text/plain; charset="us-ascii" I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net ------------------------------ Message: 2 Date: Sun, 18 Sep 2005 18:37:57 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] HT student To: , Message-ID: <1085039814-324235194@pathology.swmed.edu> Content-Type: text/plain; charset="us-ascii" At our hospital lab, anyone we hire who is registry eligible has 1 1/2 years in which to take and pass their exam (HT/HTL/CT/QIHC, etc). It is stated/written when they are hired. If they do not take AND pass within that time period, they are let go. We made it for 1.5 years, as that gives them several attempts, in case they do fail. Everyone has taken and passed. Plus, they know that once they pass the exam, they get bumped up to the next pay level. Double incentive to take the exam early and pass it. As for the commitment, I know of some labs that have offered my students to pay sign-on bonuses, moving expenses, sometimes even out-right loans, with the understanding (in writing, of course) that these are "free loans", if the person comes to work for them for 1-2 (or 3) years. If the student leaves before the 1-2-3 years, they must repay the loans - either the entire amount, or the percentage left (in other words, if they agreed to work for 2 years (24 months), and they leave after 14 months, they had 10 months that they did not fulfill the agreement, so have to pay back 10/24 of the "loan". Just some thoughts. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pruegg@ihctech.net Sent: Sunday, September 18, 2005 3:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HT student I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Sun, 18 Sep 2005 18:43:32 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] New ASCP Classification and You To: "'pam marcum'" , Message-ID: Content-Type: text/plain; charset="us-ascii" Pam - I'm curious. Who at NSH is saying this? Renee Allegruci from the ASCP Board of Registry was at the NSH S/C (at the ASCP BOR booth in the exhibit hall). She attended the Instructors of Histotechnology meeting on Monday night. I asked her, in front of about a dozen histology schools' program directors, if ASCP BOR was pursuing the lab assistant exam route, and her answer was short - "No." Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pam marcum Sent: Friday, September 16, 2005 2:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You Good Morning, I have just returned from the NSH meeting. I was surprised to find out the Histology Assistant or HA category recently presented to the NSH board and refused with a letter is moving forward. No one had told the general membership of NSH or ASCP or had it been explained to them in full. It is important for those of us who have worked so hard to get the new the education rules set in place to understand what this new registry will be and how it will affect us as professionals. This is an aid position that is OJT (on the job training) and will mean the pathologist, hospital or other private histology laboratory will be able to avoid having more than registered HT or HTL in the laboratory. The fully registered HT or HTL can then be used as the supervisor or manager and responsible for all the HA's work. This will aid in maintaining the lower pay scale we currently enjoy and help prevent us raising our overall image as professionals. Please find out more about this and respond to the ASCP/BOR with comments about this new category. The only way any of us will be heard is to let ASCP know what we think and I hope many of you will join me in letting them know we do not want to go back to the 1960s and 70s when we were all OJT and the pay is still reflecting it. (By the way I was OJT and have worked hard to improve my skills since then) We have worked hard for over thirty years to begin to be recognized as professionals not just techs! DO NOT let this stop us now so the pathologists at ASCP and AMA can continue to treat Histology as minor part of the laboratory. Histology has changed and will continue to make strides forward that require more education not less. The board of NSH did send a letter to the ASCP and tell them they would not approve this category. It is a personal feeling however, I think the Board of NSH should have notified the membership about this immediately, as a letter is not sufficient to protect us or respond as members of both NSH and ASCP. Again, write ASCP/BOR about your feelings if this important to you. Second point Cathy Locallo put a motion forward at the House of Delegates to request a Task Force to see if there is way to get histologists (HT and HTL) working for the federal government recognized as professionals and here we have a new way with the HA registry to keep us status quo. By the time the task force is in place and moving we will be down a rung again. Pamela Marcum (This is a reflection of my opinion not my employer or any other person.) Pam Marcum _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 19 Sep 2005 08:15:27 +0300 From: Saeed Al-shieban Subject: [Histonet] training in muscle biopsies handling To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 hi I am a medical tech. I am interested in handling of muscle biopsies. any body can advise me where is the best place to take training in that for 3 months at USA or UK thanks ------------------------------ Message: 5 Date: Mon, 19 Sep 2005 08:35:49 -0500 From: "Horn, Hazel V" Subject: RE: [Histonet] HT student To: pruegg@ihctech.net, Histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE21@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii Just my thoughts but you should have required that of her before she started attending school with you footing part of the bill. I think it would be inappropriate to ask/demand it now. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Sunday, September 18, 2005 5:38 PM To: pruegg@ihctech.net; histonet@pathology.swmed.edu Subject: RE: [Histonet] HT student At our hospital lab, anyone we hire who is registry eligible has 1 1/2 years in which to take and pass their exam (HT/HTL/CT/QIHC, etc). It is stated/written when they are hired. If they do not take AND pass within that time period, they are let go. We made it for 1.5 years, as that gives them several attempts, in case they do fail. Everyone has taken and passed. Plus, they know that once they pass the exam, they get bumped up to the next pay level. Double incentive to take the exam early and pass it. As for the commitment, I know of some labs that have offered my students to pay sign-on bonuses, moving expenses, sometimes even out-right loans, with the understanding (in writing, of course) that these are "free loans", if the person comes to work for them for 1-2 (or 3) years. If the student leaves before the 1-2-3 years, they must repay the loans - either the entire amount, or the percentage left (in other words, if they agreed to work for 2 years (24 months), and they leave after 14 months, they had 10 months that they did not fulfill the agreement, so have to pay back 10/24 of the "loan". Just some thoughts. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of pruegg@ihctech.net Sent: Sunday, September 18, 2005 3:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HT student I am OJT a college student working towards HT certification, I presume. She is a Jr. in college and taking all the required courses to sit for the HT/HTL exam. She works in my lab for an hourly wage while attending school. I fund a portion of her tuition costs at a local University. Is it appropriate for me to ask this person to give me a commitment to pursue HT certification when she becomes eligible? Is it also appropriate for me to ask for a commitment from this person to work in my lab after she is certified, at the going salary rate, of course? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ======================================================================== ====== ------------------------------ Message: 6 Date: Mon, 19 Sep 2005 09:13:53 -0500 From: bliven.laura@marshfieldclinic.org Subject: [Histonet] CAP Regulation To: Message-ID: <2904d01c5bd24$5d44a7e0$8e0110ac@mfldclinframe.org> The College of American Pathologists requires: "For immunohistochemistry tests that provide independent predictive/prognostic information, does the patient report include information on specimen processing, the antibody clone, and the scoring method used.?" Anyone willing to share their example? I'd like to keep it sweet, simple, and short, but have it look clean and easy to read. Please send by email or fax, void of all personal info please. Thanks, Laura Bliven Histology Lab/IHC Marshfield Laboratories 1000 N. Oak Ave. Marshfield, WI 54449 bliven.laura@marshfieldclinic.org fax#715-389-5353 ------------------------------ Message: 7 Date: Mon, 19 Sep 2005 11:15:26 -0500 From: "Jackie M O'Connor" Subject: [Histonet] Santa Cruz CD31 To: histonet@pathology.swmed.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Does anyone know if Santa Cruz ever worked out the issue with their CD31 SC1506 antibody? Last I heard, their goat was sick. Is it safe to order this antibody with a reasonable expectation that it will work? Jackie O' ------------------------------ Message: 8 Date: Mon, 19 Sep 2005 21:22:10 +1200 From: "Muhammad Tahseen" Subject: [Histonet] OCT4 antibody for immunohistochemistry. To: "HistoNet Server" Message-ID: <002401c5bcfb$9e1c5fc0$972bfea9@m7c0y4> Content-Type: text/plain; charset="iso-8859-1" Dear All, Our lab is looking for OCT4 antibody for immunohistochemistry identification of seminoma and embryonal carcinoma. Could you please provide us with the necessary information so that we can order the antibody. Thank you. Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. ------------------------------ Message: 9 Date: Mon, 19 Sep 2005 11:34:33 -0500 From: "Sebree Linda A." Subject: [Histonet] bcl-2 and bcl-6 for Ventana users To: "Histonet \(Histonet\)" Message-ID: Content-Type: text/plain; charset="us-ascii" Hello, I'm having some difficulty working up bcl-6 on our Ventana instruments and the bcl-2 we've always used isn't as robust as I'd like either. What clones and vendors are people using on their Ventana automated stainers? We have a NexES, a BenchMark and a BenchMark XT. Thanks for the info. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 ------------------------------ Message: 10 Date: Mon, 19 Sep 2005 10:49:28 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] New ASCP Classification and You To: "'Stephen Peters M.D.'" , Message-ID: <200509191649.j8JGnNfj002085@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Before we all get bent out of shape about this we need to verify that ASCP is really going forward with this classification or not. I am on the NSH BOD and at no time did we have any discussions about this with the assumption that ASCP is going forward with this. We need to ask Marilyn Gamble the NSH ASCP representative to clarify. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Saturday, September 17, 2005 8:08 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] New ASCP Classification and You I think this is a step backwards for this field. By approving of such a category they are sending the message this job is so easy you can just take anyone off the street give them a few lessons and out will pop your insitu hybridization. Your payscale will go backwards. I cannot imagine how anyone can understand what they are doing in this rather complex field without a basic knowledge of the scientific and technical principals you are using every day. Between the myriad of histochemical stains you need to know, you have now added immunohistochemistry and molecular techniques. These are very sophisticated scientific procedures. Think about the minutia of details and experience as well as technical information about your microtomes that is required just to get a nice clean unshattered ribbon of a GI biopsy. Sure, you can show someone the technical maneuvers in your free time but they will not reach nearly the same level of quality without a knowledge of the principals. As the guy reading the slides I would rather see your field require formal education and certification at all levels. I have no problem with OJ T's performing clerk duties, filing and such, but it is a bit scary to think of letting one of these loose on my biopsies or cover immunos becase every one is on vacation. My suggestion for these HA's is to designate an approved set of duties, much like doctors are given privileges for certain procedures and not others when we join a staff. Have them register with NSH as HA's in training. Make them complete classes and pass certification exams on a curriculum which is appropriate for their duties in a given time period. At least it would be like taking a correspondence course. Stephen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 22 **************************************** From Emily.Wiesner <@t> medecine.unige.ch Tue Sep 20 08:15:16 2005 From: Emily.Wiesner <@t> medecine.unige.ch (Emily Jane Wiesner-Camm) Date: Tue Sep 20 08:16:02 2005 Subject: [Histonet] Bielschowsky Staining Message-ID: <43300B64.3060403@medecine.unige.ch> Hello All. I have been staining some young rat brain sections with Bielschowsky stain, and I was wondering if anyone could tell me if the silver if deposited on only fully myelinated axons. Thank you in advance. Emily ** ** From rjbuesa <@t> yahoo.com Tue Sep 20 08:56:41 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Sep 20 08:57:02 2005 Subject: [Histonet] Tricks of the Trade. Message-ID: <20050920135641.19609.qmail@web61220.mail.yahoo.com> The Journal of Histotechnology(JOH) has a new section titled "Tricks of the Trade" (TOT) that will edit and publish those things that we all do in order to circumvent problems during grossing, cassetting, fixing, processing, cutting or staining, in order to obtain an "end product" of better quality for patient care or any research activity we may be involved with. This section cannot succeed if you are not willing to help your fellow histotechs by sending us your TOTs. Whatever you do that you know helps in your work we would like to know about. You don't have to elaborate, just a phrase containing a brief description, like: I add some drops of liquid detergent in the water bath to help the paraffin sections to expand, will be enough. After we receive your TOTs they will be sorted, classified and edited for publication in the JOH with acknowledgement of those providing the TOTs. At this moment I have been assigned the task of seeking for TOTs, and would like to receive them at my e-mail address, along with the e-mail address of each contributor so I can acknowledge receipt and keep updating about the "destiny" of each TOT. Every two weeks I intend to post in Histonet a summary of the TOTs received as an additional motivation for contributions and to share with all my fellow histotechs the TOTs we have. We at the JOH are very excited about this new feature and expect to receive your cooperation in this endeavor aimed at providing better means of work to all. Sincerely Rene J. Buesa (for the Editorial Board of the JOH). --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From contact <@t> excaliburpathology.com Tue Sep 20 09:06:05 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Tue Sep 20 09:06:30 2005 Subject: [Histonet] Sakura slide racks Message-ID: <20050920140605.300.qmail@web50312.mail.yahoo.com> There are some on Ebay right now. Do a search for Tissue Tek or Sakura. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From contact <@t> excaliburpathology.com Tue Sep 20 09:13:50 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Tue Sep 20 09:14:09 2005 Subject: [Histonet] Sakura slide racks Message-ID: <20050920141350.4775.qmail@web50311.mail.yahoo.com> Sorry, I just checked and they are no longer listed on Ebay. They were there yesterday. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From djohnson14 <@t> hotmail.com Tue Sep 20 09:33:31 2005 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Tue Sep 20 09:33:47 2005 Subject: [Histonet] Auto coverslippers In-Reply-To: Message-ID: What pricing are people seeing new and used Tape and glass coverslippers? From dpahisto <@t> yahoo.com Tue Sep 20 09:44:40 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Tue Sep 20 09:45:12 2005 Subject: [Histonet] Cutting fibers Message-ID: <20050920144440.56619.qmail@web33410.mail.mud.yahoo.com> I received a request from one of my pathologists. He will be giving me a block of tissue with an unknown fiber embedded in the tissue. He wants me to section the tissue retaining the fibers as much as possible so he can tell what the fibers are. Any suggestions would be greatly appreciated. Cindy DuBois Delta Pathology Assoc. Stockton, CA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Jeanne.Reis <@t> biogenidec.com Tue Sep 20 10:15:50 2005 From: Jeanne.Reis <@t> biogenidec.com (Jeanne Reis) Date: Tue Sep 20 10:16:17 2005 Subject: [Histonet] unattended table Message-ID: I was disappointed to see the Histonet table unattended during the recent NSH Symposium/Convention in Ft. Lauderdale. What surprised me was that even the chair was removed from behind the table on Sunday. I did find a pad of paper with names listed on it. The purpose of this????? Histonet is a great tool and not having a representative available was a lost opportunity for sharing with so many attendees. Maybe next time. From Frederick.Fifield <@t> sunhealth.org Tue Sep 20 10:38:06 2005 From: Frederick.Fifield <@t> sunhealth.org (Frederick.Fifield@sunhealth.org) Date: Tue Sep 20 10:34:59 2005 Subject: [Histonet] Re: Waste Disposal Message-ID: Our facility has a contract with a local waste disposal company which comes out quarterly to dispose of stain waste such as ammonium-silver nitrate, iron stain waste, etc. They also come out on an as needed basis to dispose of DAB and xylene. Fred Fifield BS, HT (ASCP) Pathology Section Manager Sun Health - Boswell Memorial Hospital (623) 876-5338 frederick.fifield@sunhealth.org I would like to know how everyone is disposing of the ammonium-silver nitrate waste and DAB. Any and all responses would be of great help. Thanks Annette Featherstone HT/MLT Supervisor Anatomic Pathology Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 716-859-2625 ******************************************************************************* The information contained in this transmission may be legally privileged and/or confidential information. Any dissemination, distribution or copying of this transmission by anyone other than the intended recipient is strictly prohibited. If you receive this in error, please inform the sender immediately and remove any record of this message. ******************************************************************************* For more information about Sun Health, visit us at: www.sunhealth.org From mprice26 <@t> juno.com Tue Sep 20 10:41:41 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Tue Sep 20 10:43:26 2005 Subject: [Histonet] RE:Trying to reach Gary Shackleford Message-ID: <20050920.084151.15030.519819@webmail04.nyc.untd.com> Hello Histonetters, I am trying to reach Gary Shackleford. Does anyone have contact information for him? Thank you. Marsha Price From TJJ <@t> Stowers-Institute.org Tue Sep 20 10:43:22 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Sep 20 10:43:47 2005 Subject: [Histonet] Citraconic anhydride Message-ID: My apologies in advance for those of you on both listserves... In one of my IHC workshops at the NSH, the subject of citraconic anhydride came up. Apparently a couple of people have tried it but didn't get the dramatic effects as seen in the JHC paper. Somebody mentioned one company has this commercially available, and I'm interested if this is true, which company, and any experiences using this chemical, commercial or otherwise. 'Tis a nasty chemical, and one I may well be staying away from. Best wishes, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From pruegg <@t> ihctech.net Tue Sep 20 10:48:32 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue Sep 20 10:49:00 2005 Subject: [Histonet] RE: [IHCRG] Citraconic anhydride In-Reply-To: <200509201527.j8KFRUrq018189@neo.agsci.colostate.edu> Message-ID: <200509201548.j8KFmPfj015431@chip.viawest.net> Terri, It was mentioned that one of the BioCare retrieval reagents was made with this, not sure which one, I did ask if precautions came with it and they said they did. Sounds nasty to me as well, I will stear clear. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: ihcrg-bounces@neo.agsci.colostate.edu [mailto:ihcrg-bounces@neo.agsci.colostate.edu] On Behalf Of Johnson, Teri Sent: Tuesday, September 20, 2005 8:43 AM To: Histonet Cc: ihcrg@neo.agsci.colostate.edu Subject: [IHCRG] Citraconic anhydride My apologies in advance for those of you on both listserves... In one of my IHC workshops at the NSH, the subject of citraconic anhydride came up. Apparently a couple of people have tried it but didn't get the dramatic effects as seen in the JHC paper. Somebody mentioned one company has this commercially available, and I'm interested if this is true, which company, and any experiences using this chemical, commercial or otherwise. 'Tis a nasty chemical, and one I may well be staying away from. Best wishes, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 _______________________________________________ IHCRG mailing list IHCRG@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg From pmarcum <@t> vet.upenn.edu Tue Sep 20 10:51:16 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Tue Sep 20 10:51:41 2005 Subject: [Histonet] unattended table In-Reply-To: References: Message-ID: <6.1.1.1.2.20050920114534.019910a0@mail.vet.upenn.edu> At 11:15 AM 9/20/2005, Jeanne Reis wrote: >I was disappointed to see the Histonet table unattended during the recent >NSH Symposium/Convention in Ft. Lauderdale. What surprised me was that >even the chair was removed from behind the table on Sunday. I did find a >pad of paper with names listed on it. The purpose of this????? Histonet >is a great tool and not having a representative available was a lost >opportunity for sharing with so many attendees. Maybe next time. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Jean, You now see why the VIR and HTC committees start requesting volunteers to sit at the tables prior to the meeting. They will request volunteers and have a schedule for members to sign up for specific slots. This allows coverage and commits the person to be there. I was sort of involved at the beginning of this idea and then heard nothing until just prior to the meeting. It is difficult to manage however, if the HistoNet Outpost continues it should be more formal to be sure the booth and meetings of the group are not conflicting with other NSH functions or classes. Hope this helps. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From jkiernan <@t> uwo.ca Tue Sep 20 10:56:36 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Tue Sep 20 10:57:04 2005 Subject: [Histonet] Re: Ammoniacal silver nitrate disposal References: <139141F8BAF4A642A945ECC528511AF0012A7939@kalmb02.kaleidahealth.org> Message-ID: <43303134.75E5C27D@uwo.ca> Ammoniacal silver nitrate solutions must not evaporate to dryness because the deposit contains silver azide and silver amide, a mixture known as fulminating silver that is a touch-sensitive explosive. Add some hydrochloric acid to the solution. this will precipitate all the silver as white silver chloride. This darkens on exposure to light; it is not hazardous. When you have a big bottle of silver chloride, decant off most of the supernatant (which contains ammonium chloride and dilute hydrochloric acid) and add metallic zinc to the sludge. After a few days the silver chloride changes to a heavy deposit of dark grey silver powder. Decant again and rinse with dilute hydrochloric acid to extract excess zinc, then wash the deposit with several changes of water, filter and dry. I've got two jars of silver powder made in this way, but have yet to find out who would buy it for refining. Any ideas? -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Featherstone, Annette" wrote: > > I would like to know how everyone is disposing of the ammonium-silver > nitrate waste and DAB. Any and all responses would be of great help. > Thanks > Annette Featherstone HT/MLT > Supervisor Anatomic Pathology > Kaleida Health > Buffalo General Hospital > 100 High St > Buffalo NY 14203 > 716-859-2625 > From jkiernan <@t> uwo.ca Tue Sep 20 11:05:50 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Tue Sep 20 11:06:08 2005 Subject: [Histonet] Bielschowsky Staining References: <43300B64.3060403@medecine.unige.ch> Message-ID: <4330335E.12C004DA@uwo.ca> Silver methods (if they are working properly) stain the axons, not the myelin sheaths. Unmyelinated and myelinated axons are shown. The mechanism has not been thoroughly studied for Bielschowsky-type methods, but in technically simpler procedures such as Bodian's protargol it is known that the substrate of staining is a neurofilament protein within the axoplasm (Gambetti et al. 1981 Science 213:1521-1522). -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Emily Jane Wiesner-Camm wrote: > > Hello All. > I have been staining some young rat brain sections with Bielschowsky > stain, and I was wondering if anyone could tell me if the silver if > deposited on only fully myelinated axons. > Thank you in advance. > Emily > ** > > ** > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Tue Sep 20 11:25:09 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Sep 20 11:25:40 2005 Subject: [Histonet] Re: Ammoniacal silver nitrate disposal Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175D7@lsexch.lsmaster.lifespan.org> I find it more cost effective to precipitate silver with sodium chloride - not reagent grade, just ordinary table salt from the market. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of John > A. Kiernan > Sent: Tuesday, September 20, 2005 8:56 AM > To: Featherstone, Annette > Cc: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] Re: Ammoniacal silver nitrate disposal > > Ammoniacal silver nitrate solutions must not > evaporate to dryness because the deposit contains > silver azide and silver amide, a mixture known > as fulminating silver that is a touch-sensitive > explosive. Add some hydrochloric acid to the > solution. this will precipitate all the silver > as white silver chloride. This darkens on > exposure to light; it is not hazardous. > > When you have a big bottle of silver chloride, > decant off most of the supernatant (which contains > ammonium chloride and dilute hydrochloric acid) and > add metallic zinc to the sludge. After a few days > the silver chloride changes to a heavy deposit of > dark grey silver powder. Decant again and rinse > with dilute hydrochloric acid to extract excess > zinc, then wash the deposit with several changes > of water, filter and dry. > > I've got two jars of silver powder made in this > way, but have yet to find out who would buy it > for refining. Any ideas? > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > "Featherstone, Annette" wrote: > > > > I would like to know how everyone is disposing of the ammonium-silver > > nitrate waste and DAB. Any and all responses would be of great help. > > Thanks > > Annette Featherstone HT/MLT > > Supervisor Anatomic Pathology > > Kaleida Health > > Buffalo General Hospital > > 100 High St > > Buffalo NY 14203 > > 716-859-2625 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From PMonfils <@t> Lifespan.org Tue Sep 20 11:32:24 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Sep 20 11:32:51 2005 Subject: [Histonet] Cutting fibers Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175D8@lsexch.lsmaster.lifespan.org> Hi Cindy, In case the fibers don't section well, or don't adhere to the surrounding tissue, cut some sections thicker than normal, which maximizes the chances of the fibers remaining in the section. Perhaps some sections at 5 microns, some at 10 and a few at 20 to 30 microns. Asbestos fibers for example tend to shatter when the knife hits them. But in a 20 micron section there will often be some complete fibers, the knife having passed on each side of them rather than through them. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Cindy > DuBois > Sent: Tuesday, September 20, 2005 7:44 AM > To: Histonet > Subject: [Histonet] Cutting fibers > > I received a request from one of my pathologists. He will be giving me a > block of tissue with an unknown fiber embedded in the tissue. He wants me > to section the tissue retaining the fibers as much as possible so he can > tell what the fibers are. > Any suggestions would be greatly appreciated. > Cindy DuBois > Delta Pathology Assoc. > Stockton, CA > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From d.gregg <@t> juno.com Tue Sep 20 12:15:34 2005 From: d.gregg <@t> juno.com (d.gregg@juno.com) Date: Tue Sep 20 12:22:19 2005 Subject: [Histonet] Entry level histotech position available at Plum Island Animal Disease Center Message-ID: <20050920.131534.3696.0.d.gregg@juno.com> Hi Histonetters, I posted a position about a month ago but got only high level lab managers to apply. This is a single tech lab. It is a GS 5-9 grade. Work is varied, interesting, and you will be given great freedom to manage your own work. Research emphasis is on foot-and-mouth disease pathogenesis and pathogenesis of adenovirus vectored vaccines. Most is crysectioning and IHC with some paraffin. There is some virology, necropsy assistance, and opportunities to learn laser confocal, EM, FA. and digital photomicroscopy. This unit supports one pathologist and several postdocs. Applicants need to be US citizens, have at least a BS degree, and pass a security clearance before starting work. The initial clearance takes about 3 weeks. Histo certification is not required. Training will be given on the job. Plum Island is a unique BL-3 animal research center working only on foreign diseases of animals. You will get lab clothing supplied and at least one free shower a day. It is located on the far eastern end of Long Island NY. This is a resort area surrounded but Long Island Sound and Peconic bay. The area is beautiful and rural but relatively expensive to buy real estate. There is an added pay differential to help with the higher local cost of living. It is 100 miles from NYC and 12 miles by water to Saybrook CT. Employees ride a 10 minute ferry from the tip of Long Island or a 30 minute ferry from Old Saybrook.. For anyone who is homeless due to hurricane Katrina, there is a furnished two bedroom house available for free until they can get on their feet or return to their home ( up to one year). This is not associatied with Plum Island or this job posting but is available through a local church in Southold, NY. This job will soon be posted on USAJOBS.gov. Please contact me directly for more details. Douglas Gregg DVM, PhD Veterinary pathologist Plum Island Animal Disease Center Greenport, NY, 11944 e-mail d.gregg@juno.com phone 631-323-3379 From DLGavin <@t> wyeth.com Tue Sep 20 12:32:36 2005 From: DLGavin <@t> wyeth.com (Donna L. Gavin) Date: Tue Sep 20 12:33:13 2005 Subject: [Histonet] (no subject) Message-ID: From mprice26 <@t> juno.com Tue Sep 20 12:40:48 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Tue Sep 20 12:41:39 2005 Subject: [Histonet] RE: Gary Shackleford responses Message-ID: <20050920.104104.12282.522717@webmail10.nyc.untd.com> Histonetters, Thanks to all who responded to my inquiry about Gary. I found one of his old cards and was not aware he had passed away. I hope I did not offend anyone. Marsha From Richard.Breckenridge <@t> uth.tmc.edu Tue Sep 20 13:00:48 2005 From: Richard.Breckenridge <@t> uth.tmc.edu (Breckenridge, Richard A) Date: Tue Sep 20 13:01:07 2005 Subject: [Histonet] Histotech position at UT Health Science Center Houston Medical School Message-ID: UTHSC- Houston Medical School has an opening for a Histotech II / Histotech III. Looking for someone with Renal Biopsy grossing and staining experience, as well as muscle biopsy grossing and staining experience. Competitive salary and benefits. Apply at www.uth.tmc.edu click on the "job openings" link. Thanks! Richard A. Breckenridge, HT(ASCP) Technical Director/ Chief Histology Technician UT-Houston Medical School Histology/ IHC Laboratory Office 713-500-6792 IHC Lab 713-500-5096 Histology Lab 713-500-5363 Email: Richard.Breckenridge@uth.tmc.edu From Y.Biesel <@t> web.de Tue Sep 20 13:30:02 2005 From: Y.Biesel <@t> web.de (Yvonne Biesel) Date: Tue Sep 20 13:30:41 2005 Subject: [Histonet] German technichian is looking for a new job Message-ID: <103147109@web.de> Hello Histonetters! I'm a german histotech,or to be more precise, I'm a biological Lab assistant, that worked (at this moment) in the field of Immunhohistology in a research group in New Orleans. Now, I think, everyone knows, that New Orleans and the labs there are at the moment not able to be used and so I'm looking for a new job. I'm very well experienced in Immunohistology and other histologystaining methods and also I' vesome experiences in the handling and the culture of cells. In germany I also worked a long time in a molecularbiology lab. If you are interested in a cv, please write back. your Yvonne ______________________________________________________________ Verschicken Sie romantische, coole und witzige Bilder per SMS! Jetzt bei WEB.DE FreeMail: http://f.web.de/?mc=021193 From PIXLEYSK <@t> UCMAIL.UC.EDU Tue Sep 20 13:56:25 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Tue Sep 20 13:56:53 2005 Subject: [Histonet] Fluorescent cytoplasmic marker Message-ID: Dear Histonetters: I need an antibody (or a stain) that will label all of the cytoplasm of every cell. For example, a reliable (and hopefully cheap) antibody against some common housekeeping protein that we can then localize with a fluorescent secondary antibody. I want to be able to use the confocal microscope to visualize the entire cytoplasm of every cell, because we are asking questions about where the cells are relative to an unusual substrate. We have tried staining for alpha and beta tubulin and the filamentous staining is just not sufficient. We want to fill the cytoplasm with a fluorescent marker. And we don't want to have to transfect the cells or inject them. These are mouse cells. Thanks, Sarah Pixley Univ. Cincinnati From Mf14 <@t> aol.com Tue Sep 20 14:40:16 2005 From: Mf14 <@t> aol.com (Mf14@aol.com) Date: Tue Sep 20 14:40:38 2005 Subject: [Histonet] Microtome on Ebay Message-ID: <67.4d7f05dd.3061bfa0@aol.com> If anyone out there is in need of a microtome, I just put a demo unit on e-bay. It's a model RMC MT-955. Outfitted as this microtome is, it normally would sell for over $ 6,000.00. This unit is listed at $ 250.00, and the reserve is a little over $ 1,200.00. This unit was used as a demo unit, and is in "new" condition, although I no longer have the manuals. If you would like to see additional information, you can follow this link directly to ebay and the microtome. _http://cgi.ebay.com/Microtome-Like-New-w-Quick-Release-Clamp_W0QQitemZ7547976 695QQcategoryZ26237QQrdZ1QQcmdZViewItem_ (http://cgi.ebay.com/Microtome-Like-New-w-Quick-Release-Clamp_W0QQitemZ7547976695QQcategoryZ26237QQrdZ1QQcmdZViewIt em) From Thomas.Crowell <@t> biogenidec.com Tue Sep 20 15:06:33 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Tue Sep 20 15:07:14 2005 Subject: [Histonet] Frozen sections on cryoprotected CNS In-Reply-To: Message-ID: Dear neurohistology experts, Our laboratory is experiencing great frustration in preparing 10 to 20 micron cryosections from rat and mouse brains that have been cryoprotected in 30% sucrose. The exact sequence of specimen preparation is perfusion fix with 4%PFA, drop fix in PFA for an additional 8-10 hours, transfer to 30% sucrose in PBS, before embedding in OCT. Two events occur that make sectioning difficult. 1:( the OCT does not bond well to the sample causing separation. 2 :( sections that are clearly laying flat on the cryoplate blade holder become distorted and wrinkled when thaw mounted onto the slide (Surgipath X-tra), and do not spread out well on the slide as it dries leaving lots of folds and bubbles in the tissue section. We have tried leaving the cryoprotected sample in OCT at 4C overnight to no avail, and are considering trying some gradients of sucrose, say 5%, 10%, 15% and 20%. As the neurohistolgy component to this laboratory has dramatically increased, so has our frustration level; any help or suggestions to this problem would be greatly appreciated! Thanks Tom Crowell BiogenIdec Cambridge, MA From Mf14 <@t> aol.com Tue Sep 20 15:07:56 2005 From: Mf14 <@t> aol.com (Mf14@aol.com) Date: Tue Sep 20 15:08:24 2005 Subject: [Histonet] Microtome on E-bay, Updated Bad Link Message-ID: <1e3.44634415.3061c61c@aol.com> Sorry for the bad link in the last post. This one seems to work. _http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&item=7547976695&ssPageName=ADME :B:EF:US:1_ (http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&item=7547976695&ssPageName=ADME:B:EF:US:1) From PMonfils <@t> Lifespan.org Tue Sep 20 15:24:02 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Sep 20 15:24:23 2005 Subject: [Histonet] Fluorescent cytoplasmic marker Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175D9@lsexch.lsmaster.lifespan.org> How about using a lectin instead of an antibody? They can be purchased either directly conjugated or biotinylated, so the same ABC systems, either peroxidase or fluorescent, that are used with antibodies can also be used with lectins. In my lab we do quite a bit of lectin staining, using a battery of about 35 different lectins. For those who might not be familiar with lectins, they are proteins or glycoproteins which bind strongly to specific sugars, more or less analygous to the way antibodies bind to specific proteins. Therefore they can be used to identify and localize sugars in cells and tissue sections, analygous to identifying and localizing cell proteins with antibodies. Compared with antibodies they are cheaper to buy, easier to use (no primary and secondary), less dependent on the type and extent of fixation, and yield results that are visually similar. It is also possible to use an unconjugated lectin followed by a secondary antibody against the lectin, but in my opinion that's doing it the hard way. Cytoplasm is typically rich in the sugar mannose, and the lectin Con A which binds to mannose therefore stains cytoplasm strongly. This has been consistent in my lab in many kinds of cells from a variety of species. Another good lectin for strong general cytoplasmic staining is WGA, which binds to sialic acid, another common cytoplasmic component. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Pixley, Sarah (pixleysk) > Sent: Tuesday, September 20, 2005 11:56 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Fluorescent cytoplasmic marker > > Dear Histonetters: > I need an antibody (or a stain) that will label all of the cytoplasm of > every cell. For example, a reliable (and hopefully cheap) antibody > against some common housekeeping protein that we can then localize with > a fluorescent secondary antibody. I want to be able to use the confocal > microscope to visualize the entire cytoplasm of every cell, because we > are asking questions about where the cells are relative to an unusual > substrate. We have tried staining for alpha and beta tubulin and the > filamentous staining is just not sufficient. We want to fill the > cytoplasm with a fluorescent marker. And we don't want to have to > transfect the cells or inject them. These are mouse cells. > Thanks, > Sarah Pixley > Univ. Cincinnati > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From pmarcum <@t> vet.upenn.edu Tue Sep 20 15:29:10 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Tue Sep 20 15:29:36 2005 Subject: [Histonet] Free Ultra Microtome for Shipping Cost Message-ID: <6.1.1.1.2.20050920162345.0194d8b0@mail.vet.upenn.edu> Good Afternoon HistoNet, We have two microtome here at the University that we going to be placed in the dump until our head of Pathology asked me to take a look. We have one LKB Historange (fairly complete) and one I am not sure about so the historians out there may know. It is a Riechert Type 709901. I also have a Sorvall MT2 that needs a glass/diamond knife holder but appears to run otherwise. If anyone would like any of these instruments for use or parts they are free if you pay the shipping to you. Just let me know and he prefers they go to laboratory not a company although I think we could negotiate if need be. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From mtitford <@t> aol.com Tue Sep 20 16:00:26 2005 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Tue Sep 20 16:00:50 2005 Subject: [Histonet] Histonet table Message-ID: <8C78C3D75F13568-618-3EA@MBLK-M26.sysops.aol.com> I attended the meeting at the NSH convention. The Histonet table had a sign up sheet. Later a notice appeared announcing a social meeting at about 6.20 at the Marina Marriott hotel across the road. That was before the Thermo party. I attended the early part of that meeting. A firm had offered to sponsor that meeting and will sponsor Histonet meetings at future NSH conventions. Their representative was there. I think that is an adequate response to what is a low key information exchange system that our Histonet system is, especially when you consider how busy everyone is at the convention. I would like to thank Rebecca Orr for organizing the Histonet table. Mike Titford Pathology USA Mobile AL From cforster <@t> umn.edu Tue Sep 20 16:28:14 2005 From: cforster <@t> umn.edu (Colleen Forster) Date: Tue Sep 20 16:28:34 2005 Subject: [Histonet] Histonet table In-Reply-To: <8C78C3D75F13568-618-3EA@MBLK-M26.sysops.aol.com> References: <8C78C3D75F13568-618-3EA@MBLK-M26.sysops.aol.com> Message-ID: <43307EEE.1080308@umn.edu> Yes, and we hope that her next year is a good one and we will see her at the meeting. Colleen Forster mtitford@aol.com wrote: >I attended the meeting at the NSH convention. The Histonet table had a sign up sheet. Later a notice appeared announcing a social meeting at about 6.20 at the Marina Marriott hotel across the road. That was before the Thermo party. I attended the early part of that meeting. A firm had offered to sponsor that meeting and will sponsor Histonet meetings at future NSH conventions. Their representative was there. > >I think that is an adequate response to what is a low key information exchange system that our Histonet system is, especially when you consider how busy everyone is at the convention. > >I would like to thank Rebecca Orr for organizing the Histonet table. > >Mike Titford >Pathology >USA Mobile AL >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >. > > > From llewllew <@t> shaw.ca Tue Sep 20 16:45:22 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Tue Sep 20 16:46:20 2005 Subject: [Histonet] Fluorescent cytoplasmic marker References: <09C945920A6B654199F7A58A1D7D1FDE017175D9@lsexch.lsmaster.lifespan.org> Message-ID: <001e01c5be2c$9a92fd50$7e034246@yourlk4rlmsu> Would eosin Y do? It fluoresces green. Bryan Llewellyn >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of >> Pixley, Sarah (pixleysk) >> Sent: Tuesday, September 20, 2005 11:56 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Fluorescent cytoplasmic marker >> >> Dear Histonetters: >> I need an antibody (or a stain) that will label all of the cytoplasm of >> every cell. For example, a reliable (and hopefully cheap) antibody >> against some common housekeeping protein that we can then localize with >> a fluorescent secondary antibody. I want to be able to use the confocal >> microscope to visualize the entire cytoplasm of every cell, because we >> are asking questions about where the cells are relative to an unusual >> substrate. We have tried staining for alpha and beta tubulin and the >> filamentous staining is just not sufficient. We want to fill the >> cytoplasm with a fluorescent marker. And we don't want to have to >> transfect the cells or inject them. These are mouse cells. >> Thanks, >> Sarah Pixley >> Univ. Cincinnati >> >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jengirl1014 <@t> yahoo.com Tue Sep 20 23:48:06 2005 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Tue Sep 20 23:48:29 2005 Subject: [Histonet] New baby Message-ID: <20050921044806.68533.qmail@web60624.mail.yahoo.com> I thought I'd share my joy with all of you. On September 1st, I had a little girl. Her name is Abigail Paige (Abbi for short) and she weighed in at 9 lbs. and 8.2 oz (43.8 kg I believe that is). All is well and her big sister (Michaela is going to be 4 next month) is very happy to have a new play mate! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 410-955-9688 e-mail: jengirl1014@yahoo.com __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From PKamalavenkatesh <@t> wockhardtin.com Tue Sep 20 23:54:26 2005 From: PKamalavenkatesh <@t> wockhardtin.com (PKamalavenkatesh@wockhardtin.com) Date: Tue Sep 20 23:51:21 2005 Subject: [Histonet] rnithine carbamoyltransferase detection in RAT liver Message-ID: Dear All, We need to standardize simple histochemical detection Ornithine Carbamyl Transferase in rat liver. We got some references which are mentioning a technique called "Mizutani technique for ornithine carbamoyltransferase detection in liver". If you are having any idea regarding this, we are in need of it. Regards kamalavenkatesh Wockhardt Reserch Center New Drug Discovery - Preclinical safety Evaluation India --------------------- Disclaimer ------------------------------------------------------------------------------ Information transmitted by this E-MAIL is proprietary to Wockhardt Ltd. and/ or its Group Companies and/or,its Customers and is intended for use only by the individual or entity to which it is addressed, and may contain information that is privileged, confidential or exempt from disclosure under applicable law. If you are not the intended recipient or it appears that this mail has been forwarded to you without proper authority, you are notified that any use or dissemination of this information in any manner is strictly prohibited. In such cases, please delete this mail from your records. --------------------------------------------------------------------------------------------------------------- From Krat18 <@t> aol.com Wed Sep 21 00:43:25 2005 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Wed Sep 21 00:44:01 2005 Subject: [Histonet] New baby Message-ID: <1d7.454bcf49.30624cfd@aol.com> Congratulations to you all! It's great to hear joyful news occasionally! Thank you for sharing your happiness with us! Karen Raterman From Gunilla.Aronsson <@t> odontologi.gu.se Wed Sep 21 03:47:47 2005 From: Gunilla.Aronsson <@t> odontologi.gu.se (Gunilla Aronsson) Date: Wed Sep 21 03:49:45 2005 Subject: [Histonet] Fluorescent mitochondria marker Message-ID: <623B14C0-2A7C-11DA-9E3A-000393BD4D72@odontologi.gu.se> Dear Histonetters, I need an antibody or stain that will label all mitochondria in human fibroblasts. I want to be able to use the confocal microscope to visualize the amount of mitochondria in the cells. Thanks Gunilla Aronsson G?teborg University From cormier <@t> MIT.EDU Wed Sep 21 04:42:39 2005 From: cormier <@t> MIT.EDU (Kathy Cormier) Date: Wed Sep 21 04:43:31 2005 Subject: [Histonet] RE: [IHCRG] Citraconic anhydride In-Reply-To: <200509201548.j8KFmPfj015431@chip.viawest.net> References: <200509201527.j8KFRUrq018189@neo.agsci.colostate.edu> Message-ID: <5.2.1.1.2.20050921053451.011a0598@po14.mit.edu> Hello All, We have used this a few times here. We have not had the same significant difference in staining intensity that the authors of the paper had. This may be in large part to the "newbie tech" who performed the IHC here. I think in more experienced hands that this probably will work better. It is nasty stuff though, so be very careful. I did call Biocares' tech services and they claimed that none of the retrieval soln's that they have contain this chemical. Perhaps someone else carries it? Kathy Cormier Histology Necropsy Supervisor Division of Comparative Medicine MIT At 09:48 AM 9/20/2005 -0600, Patsy Ruegg wrote: >Terri, >It was mentioned that one of the BioCare retrieval reagents was made with >this, not sure which one, I did ask if precautions came with it and they >said they did. Sounds nasty to me as well, I will stear clear. >Patsy > > >Patsy Ruegg, HT(ASCP)QIHC >IHCtech, LLC >Fitzsimmons BioScience Park >12635 Montview Blvd. Suite 216 >Aurora, CO 80010 >P-720-859-4060 >F-720-859-4110 >wk email pruegg@ihctech.net >web site www.ihctech.net > > >This email is confidential and intended solely for the use of the Person(s) >('the intended recipient') to whom it was addressed. Any views or opinions >presented are solely those of the author. It may contain information that is >privileged & confidential within the meaning of applicable law. Accordingly >any dissemination, distribution, copying, or other use of this message, or >any of its contents, by any person other than the intended recipient may >constitute a breach of civil or criminal law and is strictly prohibited. If >you are NOT the intended recipient please contact the sender and dispose of >this e-mail as soon as possible. > > >-----Original Message----- >From: ihcrg-bounces@neo.agsci.colostate.edu >[mailto:ihcrg-bounces@neo.agsci.colostate.edu] On Behalf Of Johnson, Teri >Sent: Tuesday, September 20, 2005 8:43 AM >To: Histonet >Cc: ihcrg@neo.agsci.colostate.edu >Subject: [IHCRG] Citraconic anhydride > >My apologies in advance for those of you on both listserves... > >In one of my IHC workshops at the NSH, the subject of citraconic anhydride >came up. Apparently a couple of people have tried it but didn't get the >dramatic effects as seen in the JHC paper. Somebody mentioned one company >has this commercially available, and I'm interested if this is true, which >company, and any experiences using this chemical, commercial or otherwise. >'Tis a nasty chemical, and one I may well be staying away from. > >Best wishes, > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 > >_______________________________________________ >IHCRG mailing list >IHCRG@neo.agsci.colostate.edu >http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg > >_______________________________________________ >IHCRG mailing list >IHCRG@neo.agsci.colostate.edu >http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg From anders.lewen <@t> neurokir.uu.se Wed Sep 21 06:12:02 2005 From: anders.lewen <@t> neurokir.uu.se (Anders Lewen) Date: Wed Sep 21 06:16:32 2005 Subject: [Histonet] New baby In-Reply-To: <20050921044806.68533.qmail@web60624.mail.yahoo.com> References: <20050921044806.68533.qmail@web60624.mail.yahoo.com> Message-ID: <1127301122.4331400293f3a@webmail.uu.se> I'm happy for you Jennifer. But you must have had a terrible delivery since she is weighing 43.8 kg!!!!!!!!!! (about 70 lbs...) Good luck! Anders Citerar Jennifer Sipes : > I thought I'd share my joy with all of you. On September 1st, I had a little > girl. Her name is Abigail Paige (Abbi for short) and she weighed in at 9 > lbs. and 8.2 oz (43.8 kg I believe that is). All is well and her big sister > (Michaela is going to be 4 next month) is very happy to have a new play > mate! > > > Jennifer K. Sipes, RALAT > Sr. Laboratory Technician > Johns Hopkins University > Ross 929 > 720 Rutland Avenue > Baltimore, MD 21205 > phone: 410-614-0131 > 410-955-9688 > e-mail: jengirl1014@yahoo.com > > > > > > > > > > > > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ljb <@t> medicine.wisc.edu Wed Sep 21 08:27:00 2005 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Wed Sep 21 08:27:33 2005 Subject: [Histonet] New baby Message-ID: OUCH!!!! >>> Anders Lewen 09/21/05 6:12 AM >>> I'm happy for you Jennifer. But you must have had a terrible delivery since she is weighing 43.8 kg!!!!!!!!!! (about 70 lbs...) Good luck! Anders Citerar Jennifer Sipes : > I thought I'd share my joy with all of you. On September 1st, I had a little > girl. Her name is Abigail Paige (Abbi for short) and she weighed in at 9 > lbs. and 8.2 oz (43.8 kg I believe that is). All is well and her big sister > (Michaela is going to be 4 next month) is very happy to have a new play > mate! > > > Jennifer K. Sipes, RALAT > Sr. Laboratory Technician > Johns Hopkins University > Ross 929 > 720 Rutland Avenue > Baltimore, MD 21205 > phone: 410-614-0131 > 410-955-9688 > e-mail: jengirl1014@yahoo.com > > > > > > > > > > > > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Wed Sep 21 08:40:47 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Sep 21 08:41:31 2005 Subject: [Histonet] New baby Message-ID: It doesn't matter if they're 5lbs or 12lbs, they all feel like 70.......... "LaCinda Burchell" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/21/2005 08:27 AM To: , cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] New baby OUCH!!!! >>> Anders Lewen 09/21/05 6:12 AM >>> I'm happy for you Jennifer. But you must have had a terrible delivery since she is weighing 43.8 kg!!!!!!!!!! (about 70 lbs...) Good luck! Anders Citerar Jennifer Sipes : > I thought I'd share my joy with all of you. On September 1st, I had a little > girl. Her name is Abigail Paige (Abbi for short) and she weighed in at 9 > lbs. and 8.2 oz (43.8 kg I believe that is). All is well and her big sister > (Michaela is going to be 4 next month) is very happy to have a new play > mate! > > > Jennifer K. Sipes, RALAT > Sr. Laboratory Technician > Johns Hopkins University > Ross 929 > 720 Rutland Avenue > Baltimore, MD 21205 > phone: 410-614-0131 > 410-955-9688 > e-mail: jengirl1014@yahoo.com > > > > > > > > > > > > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marirose.Satterfield <@t> MercyMemorial.org Wed Sep 21 08:54:53 2005 From: Marirose.Satterfield <@t> MercyMemorial.org (Satterfield, Marirose) Date: Wed Sep 21 08:55:15 2005 Subject: [Histonet] cryostat decontamination Message-ID: <545CCCF1899E2042AA1C7A0DF101FC7C137359@mmh-01sv0101ew.mercymemorial.org> CAP Checklist Question ANP.24250 Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontaminated records evident? They note that the interior of the cryostat can be decontaminated with 70% ethanol. Later on they say it should be defrosted and decontaminated with a tuberculocidal disinfectant at an interval appropriate for the institution. Are they suggesting that after each use it get decontaminated with the 70% ethanol and only periodically shut down unit and use the TB disinfectant? Thanks M Satterfield From vazquezr <@t> ohsu.edu Wed Sep 21 09:03:39 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Sep 21 09:04:21 2005 Subject: [Histonet] New baby Message-ID: what did you eat?!!!! >>> "LaCinda Burchell" 9/21/2005 6:27 AM >>> OUCH!!!! >>> Anders Lewen 09/21/05 6:12 AM >>> I'm happy for you Jennifer. But you must have had a terrible delivery since she is weighing 43.8 kg!!!!!!!!!! (about 70 lbs...) Good luck! Anders Citerar Jennifer Sipes : > I thought I'd share my joy with all of you. On September 1st, I had a little > girl. Her name is Abigail Paige (Abbi for short) and she weighed in at 9 > lbs. and 8.2 oz (43.8 kg I believe that is). All is well and her big sister > (Michaela is going to be 4 next month) is very happy to have a new play > mate! > > > Jennifer K. Sipes, RALAT > Sr. Laboratory Technician > Johns Hopkins University > Ross 929 > 720 Rutland Avenue > Baltimore, MD 21205 > phone: 410-614-0131 > 410-955-9688 > e-mail: jengirl1014@yahoo.com > > > > > > > > > > > > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Wed Sep 21 09:08:38 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Sep 21 09:09:28 2005 Subject: [Histonet] cryostat decontamination Message-ID: I shut down the cryo and decontaminate with Santimaster IV which is a broad spectrum and non corrosive, I leave it on for about 10 minutes, then I use a 70% ethanol, and then I use an absolute. Robyn OHSU >>> "Satterfield, Marirose" 9/21/2005 6:54 AM >>> CAP Checklist Question ANP.24250 Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontaminated records evident? They note that the interior of the cryostat can be decontaminated with 70% ethanol. Later on they say it should be defrosted and decontaminated with a tuberculocidal disinfectant at an interval appropriate for the institution. Are they suggesting that after each use it get decontaminated with the 70% ethanol and only periodically shut down unit and use the TB disinfectant? Thanks M Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anders.lewen <@t> neurokir.uu.se Wed Sep 21 10:34:01 2005 From: anders.lewen <@t> neurokir.uu.se (Anders Lewen) Date: Wed Sep 21 10:38:48 2005 Subject: [Histonet] New baby In-Reply-To: References: Message-ID: <1127316841.43317d69dbfa3@webmail.uu.se> Actually, I feel Jennifer should answer this important question - maybe she misplaced the lb decimal! and it is a 90+lb baby. I think we owe her credit for what she delivered! Anders Citerar: I think poor Jennifer misplaced her decimal point however after passing a 9+lb baby who can blame her. 43.8k would actually be 96.8lb (all us histotechs should know our metric conversions). Congratulations Jennifer and I wish you a speedy recovery from your "ordeal". Chuck Embrey Citerar Robyn Vazquez : > what did you eat?!!!! > > >>> "LaCinda Burchell" 9/21/2005 6:27 AM >>> > OUCH!!!! > > >>> Anders Lewen 09/21/05 6:12 AM >>> > I'm happy for you Jennifer. But you must have had a terrible delivery since > she > is weighing 43.8 kg!!!!!!!!!! (about 70 lbs...) > Good luck! > Anders > > > Citerar Jennifer Sipes : > > > I thought I'd share my joy with all of you. On September 1st, I had a > little > > girl. Her name is Abigail Paige (Abbi for short) and she weighed in at 9 > > lbs. and 8.2 oz (43.8 kg I believe that is). All is well and her big > sister > > (Michaela is going to be 4 next month) is very happy to have a new play > > mate! > > > > > > Jennifer K. Sipes, RALAT > > Sr. Laboratory Technician > > Johns Hopkins University > > Ross 929 > > 720 Rutland Avenue > > Baltimore, MD 21205 > > phone: 410-614-0131 > > 410-955-9688 > > e-mail: jengirl1014@yahoo.com > > > > > > > > > > > > > > > > > > > > > > > > > > __________________________________________________ > > Do You Yahoo!? > > Tired of spam? Yahoo! Mail has the best spam protection around > > http://mail.yahoo.com > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From sjchtascp <@t> yahoo.com Wed Sep 21 11:03:21 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Sep 21 11:04:01 2005 Subject: [Histonet] frozen section trichrome Message-ID: <20050921160321.47984.qmail@web90208.mail.scd.yahoo.com> I tried a gomori's trichrome on 2 FS tissues this morning, rat muscle and rat liver. The liver was exceptable although the smooth muscle was much more purlpe then on paraffin sections. The muscle was opposite what it should have been, with green muscle and red collagen. Heres the procedure I followed: 10 min in NBF, 5 min in Gills3, 15 min gomoris, rinse in 0.5% acetic water. Steve --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From ROrr <@t> enh.org Wed Sep 21 11:30:46 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Wed Sep 21 11:31:12 2005 Subject: [Histonet] CAP ? on the histonet Message-ID: Laura!! Are you still looking for disclaimers? I have some I can share with you, Would you mind sending me your fax number again? I'd appreciate it if you could give me the regulation number, so I can check with our Docs...on it... What I can tell you is that we've always passed with flying colors with what we have.... You'll see we reference the Her2 product but do not specifically state anything about the processing...but we interpret that with the general statement when we say "performance characteristics determined..." Let me know if you still want it I'll fax it right over! How's the biocare oven? I love mine! THx, Becky Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 From celebrej <@t> HHSC.CA Wed Sep 21 11:37:09 2005 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Wed Sep 21 11:36:59 2005 Subject: [Histonet] CJD protocols anyone? Message-ID: <3AADFB88753AD31189C100902786B91C14E6E297@hch_nt_exchange.hhsc.ca> Hi there Histoland! Our department has been hit by several 'query' CJD surgical cases lately, since we rarely handled any questionable cases in the past we are trying to prepare an updated protocol that will include the handling of these specimens from begining to end. I would appreciate hearing from anyone who deals with CJD cases from safety protocols, protocols on deactivation, processing, cutting and staining, everything and anything dealing with CJD. Thanks Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca The Cornerstone of Care Campaign for Hamilton Health Sciences needs your support. Please, visit hamiltonhealthsciences.ca today and help make something great even greater. This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From PIXLEYSK <@t> UCMAIL.UC.EDU Wed Sep 21 12:47:23 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Wed Sep 21 12:47:42 2005 Subject: [Histonet] RE: cutting brain sections on the cryostat Message-ID: Dear Thomas Crowell: I am not sure exactly what might be your problem, but I can tell you what we do that is working, and may be slight changes compared to what you are doing. First, after perfusion with 4% paraformaldehyde in 0.1 M phosphate buffer, we only postfix for 2-4 hours. Then we put the tissue (rat or mouse brain) into 30% sucrose in 0.1 M phosphate buffer (PB) and leave it for at least overnight (you don't say how long you leave it) or until the tissue floats. I don't know if the PBS, with saline, vs. PB, or the timing makes any difference. Then we use the Shandon (now Thermo) M1 embedding matrix instead of OCT. We have found the M1 to be preferable to OCT in that there is less curling of the sections, better ease of cutting. I would not leave the tissue in either matrix for very long; it is not particularly good for the tissue. Your OCT compound may also have gone bad. We have had the M1 matrix go bad after repeated use, in and out of the refrigerator. When it goes bad, it does not freeze well and remains soft and mushy around the frozen tissue. OCT may do something else when bad. Your freezing method may also complicate matters. We have had trouble using chilled isopentane with brains because the tissue cracks. Thus, I speculate that it may possibly also cause contraction, away from the OCT. There were some recent good threads on this board about freezing methods--I would recommend checking for those. We now freeze the tissue in the cryostat by putting the tissue on the chuck and then adding dry ice powder onto the tissue. Less cracking/distortion, and no other problems so far. However, we haven't recently done whole brain that way, only smaller slices. Other than that, you might want to check your cryostat. The settings for the knife angle, the temperature, the roll blade can all affect how the sections come off and adhere to the slide. Good luck! Sarah Pixley (p.s. thanks to all who are answering my question about fluorescent cytoplasmic markers--I am going to check your suggestions out!) From tmmrosla <@t> healtheast.org Wed Sep 21 12:45:03 2005 From: tmmrosla <@t> healtheast.org (Mrosla, Tina M) Date: Wed Sep 21 13:39:09 2005 Subject: [Histonet] plants in histology lab Message-ID: <22E6C4C32AFB3E44B7056C7EC3E1F204DC474F@HECLUSTER.HEALTHEAST.LOC> We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. From victor <@t> pathology.washington.edu Wed Sep 21 13:48:43 2005 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Wed Sep 21 13:49:14 2005 Subject: [Histonet] plants in histology lab In-Reply-To: <22E6C4C32AFB3E44B7056C7EC3E1F204DC474F@HECLUSTER.HEALTHEAST.LOC> References: <22E6C4C32AFB3E44B7056C7EC3E1F204DC474F@HECLUSTER.HEALTHEAST.LOC> Message-ID: <4331AB0B.7040005@pathology.washington.edu> Spider plants seem to like the fumes and I'm all for plants in the lab unless there are other regulations. Victor Mrosla, Tina M wrote: >We are trying to decide if we should have plants in our histology lab or not. >It would be nice, for the formalin fumes and such, but we are wondering about contamination. >Does anyone have any suggestions and would certain plants be better to have than others, or none at all? >Thank you, >Tina > >St. Joseph's Histology >St. Paul, MN > > > >The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From Sue.Kapoor <@t> uhsi.org Wed Sep 21 13:50:34 2005 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Wed Sep 21 13:51:06 2005 Subject: [Histonet] Retic Stain Kit Message-ID: <61E9F2400F53D5119CFC00508B44E33B02E9E1A1@khmcexch.uhsi.org> Hi everyone, We currently use Snook's method for reticulum which uses uranium nitrate and I'd really like to get rid of using it. I'm wondering if anyone is using a commerically bought retic stain kit and would appreciate your comments. Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From MICHAEL.OWEN <@t> fda.gov Wed Sep 21 13:54:50 2005 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Wed Sep 21 13:55:33 2005 Subject: [Histonet] Plants in Histology Lab Message-ID: Dear Dr. Mrosla, According to the references below, decorative plants are restricted from use at BSL-3 and higher. Plants (and animals not involved in experiments) harbor their own microbial flora, which could contaminate the work or even infect the worker; thus it is prudent to prohibit their use in all microbiological laboratories. Biosafety in Microbiological and Biomedical Laboratories Fourth Edition (BMBL) http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm U.S. Department of Health and Human Services Centers for Disease Control and Prevention National Institutes of Health May 1999 NIH Guidelines for Recombinant DNA and Gene Transfer http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html U.S. Department of Health and Human Services National Institutes of Health April 2002 WHO Biological Safety Manual Third Edition http://www.who.int/csr/resources/publications/biosafety/WHO_CDS_CSR_LYO_2004 _11/en World Health Organization December 2004 Biological Safety Principles and Practices Third Edition Edited by Diane O. Fleming and Debra L. Hunt American Society of Microbiology December 2000 I hope this information is helpful (or makes you laugh if it is not useful :o) ). Sincerely, The FDA Lab Rat Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From JWEEMS <@t> sjha.org Wed Sep 21 14:00:45 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Sep 21 14:01:06 2005 Subject: [Histonet] plants in histology lab Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01304CE3@sjhaexc02.sjha.org> Airplane/spider plants are good ones that absord fumes, and seem to live through anything. Also, philadindrin do well - grow all ther place. If you're in a hospital lab, it's a good thing. No sure about research and GLP regs and such. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mrosla, Tina M Sent: Wednesday, September 21, 2005 1:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] plants in histology lab We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Stephen.J.Scholz <@t> osfhealthcare.org Wed Sep 21 14:00:47 2005 From: Stephen.J.Scholz <@t> osfhealthcare.org (Scholz, Stephen J.) Date: Wed Sep 21 14:01:24 2005 Subject: [Histonet] plants in histology lab Message-ID: <7F1312711CA7474A89B3DF8BA0BA54D01333B2@pmc-rfd-mx01.intranet.osfnet.org> Tina; Plants in the lab are great. Are you concerned about the plant contaminating the lab? If so, I have never heard of it. (I wouldn't put it directly over my waterbath) As far as what plants to use I believe that NASA did a study on the affect of plants to indoor air quality and found that the philodendron, spider plant and the golden pothos plants were the best at reducing formaldehyde. I'm sure there are a few others that would also be helpful. Happy gardening; steve -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mrosla, Tina M Sent: Wednesday, September 21, 2005 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] plants in histology lab We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Wed Sep 21 14:05:21 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Sep 21 14:05:41 2005 Subject: [Histonet] plants in histology lab Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01304CE4@sjhaexc02.sjha.org> Now I proof read! That would be philodendron I believe, and "grow all over the place"! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Wednesday, September 21, 2005 3:01 PM To: Mrosla, Tina M; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] plants in histology lab Airplane/spider plants are good ones that absord fumes, and seem to live through anything. Also, philadindrin do well - grow all ther place. If you're in a hospital lab, it's a good thing. No sure about research and GLP regs and such. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mrosla, Tina M Sent: Wednesday, September 21, 2005 1:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] plants in histology lab We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From info <@t> instrumedics.com Wed Sep 21 14:11:04 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Wed Sep 21 14:12:27 2005 Subject: [Histonet] Frozen sections on cryoprotected CNS References: Message-ID: <008101c5bee0$4f027500$6401a8c0@INSTRUMEDICS22> Thomas, I think that you may solve your problems with the CryoJane Tape-Transfer process. The section is captured on a cold special tape as it is cut .It is flat, unfolded and intact. It is then transferred to a special cold adhesive coated slide. The adhesive is polymerized with a 8 millisecond flash. The tape is peeled away inside the cryostat.. The section remains frozen during the entire process preserving the morphology. The section is finally melted in a fixative where the melting of the ice and the fixation happen simultaneously or by freeze substitution where the ice dissolves(no melting) in cold acetone. The section is "dry". The best preservation of morphological detail is seen with freeze-substitution. The caveat is that the tissue must be snap frozen so that the ice crystal growth is minimal. Please visit our web for all the details www.instrumedics.com Bernice 800-237-2772 ----- Original Message ----- From: "Thomas Crowell" To: Sent: Tuesday, September 20, 2005 4:06 PM Subject: [Histonet] Frozen sections on cryoprotected CNS > Dear neurohistology experts, > > Our laboratory is experiencing great frustration in preparing 10 to 20 > micron cryosections from rat and mouse brains that have been cryoprotected > in 30% sucrose. The exact sequence of specimen preparation is perfusion > fix with 4%PFA, drop fix in PFA for an additional 8-10 hours, transfer to > 30% sucrose in PBS, before embedding in OCT. Two events occur that make > sectioning difficult. 1:( the OCT does not bond well to the sample > causing separation. 2 :( sections that are clearly laying flat on the > cryoplate blade holder become distorted and wrinkled when thaw mounted > onto the slide (Surgipath X-tra), and do not spread out well on the slide > as it dries leaving lots of folds and bubbles in the tissue section. > > We have tried leaving the cryoprotected sample in OCT at 4C overnight to > no avail, and are considering trying some gradients of sucrose, say 5%, > 10%, 15% and 20%. As the neurohistolgy component to this laboratory has > dramatically increased, so has our frustration level; any help or > suggestions to this problem would be greatly appreciated! > > Thanks > Tom Crowell > BiogenIdec > Cambridge, MA > From funderwood <@t> mcohio.org Wed Sep 21 14:16:30 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Wed Sep 21 14:17:08 2005 Subject: [Histonet] plants in histology lab Message-ID: I've had a bonsai tree in my lab for the past 5 years and it's doing great. Hard to say if the fumes contribute to it's contorted appearance. Fred >>> "Mrosla, Tina M" 09/21/05 01:45PM >>> We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Wed Sep 21 14:26:22 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Wed Sep 21 14:26:48 2005 Subject: [Histonet] plants in histology lab In-Reply-To: References: Message-ID: <6.1.1.1.2.20050921152516.019eb910@mail.vet.upenn.edu> Be careful Fred, they may try to take that further for those of us who have been in Histology for a long time!! Pam Marcum At 03:16 PM 9/21/2005, Fred Underwood wrote: >I've had a bonsai tree in my lab for the past 5 years and it's doing >great. Hard to say if the fumes contribute to it's contorted >appearance. > >Fred > > >>> "Mrosla, Tina M" 09/21/05 01:45PM >>> >We are trying to decide if we should have plants in our histology lab >or not. >It would be nice, for the formalin fumes and such, but we are wondering >about contamination. >Does anyone have any suggestions and would certain plants be better to >have than others, or none at all? >Thank you, >Tina > >St. Joseph's Histology >St. Paul, MN > > > >The information included in this e-mail message, including any >attachments, is intended only for the person or organization to which it >is addressed. This e-mail message may contain information that is >privileged or confidential. If you receive this e-mail message and are >not the intended recipient or responsible for delivering the message to >the intended recipient, you may not use, disseminate, distribute or copy >the information included in this e-mail and any attachments. If you >received this e-mail message by mistake, please reply by e-mail and >destroy all copies of this message and any attachments. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Wed Sep 21 14:44:30 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Wed Sep 21 14:46:06 2005 Subject: [Histonet] plants in histology lab Message-ID: We have a Norfolk pine that is thriving. We tell people that come to visit that we feed it placenta parts. Grosses them out and they don't touch it. Linda >>> "Pamela Marcum" 09/21/2005 3:26:22 PM >>> Be careful Fred, they may try to take that further for those of us who have been in Histology for a long time!! Pam Marcum At 03:16 PM 9/21/2005, Fred Underwood wrote: >I've had a bonsai tree in my lab for the past 5 years and it's doing >great. Hard to say if the fumes contribute to it's contorted >appearance. > >Fred > > >>> "Mrosla, Tina M" 09/21/05 01:45PM >>> >We are trying to decide if we should have plants in our histology lab >or not. >It would be nice, for the formalin fumes and such, but we are wondering >about contamination. >Does anyone have any suggestions and would certain plants be better to >have than others, or none at all? >Thank you, >Tina > >St. Joseph's Histology >St. Paul, MN > > > >The information included in this e-mail message, including any >attachments, is intended only for the person or organization to which it >is addressed. This e-mail message may contain information that is >privileged or confidential. If you receive this e-mail message and are >not the intended recipient or responsible for delivering the message to >the intended recipient, you may not use, disseminate, distribute or copy >the information included in this e-mail and any attachments. If you >received this e-mail message by mistake, please reply by e-mail and >destroy all copies of this message and any attachments. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Wed Sep 21 14:53:06 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Sep 21 14:53:38 2005 Subject: [Histonet] plants in histology lab Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175DC@lsexch.lsmaster.lifespan.org> Never had plants in the lab. However, years ago I worked in a lab with rather poor ventilation, and we used to open a couple of windows to let some fresh air in. One day while checking the finished slides before they went out to the pathologists, I noticed a strange object on a few different slides. They looked sort of like little starfish, which is what we ended up calling them. Some of them were stained with the same dyes that the tissue was stained with, while some of them appeared to be unstained. Over the next few days I noticed additional and more frequent examples of this contaminant. To make a long story short, this turned out to be pollen from some tree outside the lab, which was coming in through the open windows. It was settling on the freshly cut slides before they went into the drying oven, and apparently was also getting onto the slides during coverslipping. I wiped the bench near the windows with a moist wipe, rinsed it off in a little water and spun it down. There were hundreds of these tiny particles all over the bench. After that we kept the windows closed, at least in the springtime! I don't know if plants in the lab might cause such a problem. Presumably non-flowering plants wouldn't (then again, ferns do produce spores!). But it's something to consider. From Jackie.O'Connor <@t> abbott.com Wed Sep 21 15:10:26 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Sep 21 15:11:09 2005 Subject: [Histonet] plants in histology lab Message-ID: Did you name it Audrey? Perhaps Audrey II? "Linda Blazek" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/21/2005 02:44 PM To: tmmrosla@healtheast.org, histonet@lists.utsouthwestern.edu, funderwood@mcohio.org, pmarcum@vet.upenn.edu cc: Subject: Re: [Histonet] plants in histology lab We have a Norfolk pine that is thriving. We tell people that come to visit that we feed it placenta parts. Grosses them out and they don't touch it. Linda >>> "Pamela Marcum" 09/21/2005 3:26:22 PM >>> Be careful Fred, they may try to take that further for those of us who have been in Histology for a long time!! Pam Marcum At 03:16 PM 9/21/2005, Fred Underwood wrote: >I've had a bonsai tree in my lab for the past 5 years and it's doing >great. Hard to say if the fumes contribute to it's contorted >appearance. > >Fred > > >>> "Mrosla, Tina M" 09/21/05 01:45PM >>> >We are trying to decide if we should have plants in our histology lab >or not. >It would be nice, for the formalin fumes and such, but we are wondering >about contamination. >Does anyone have any suggestions and would certain plants be better to >have than others, or none at all? >Thank you, >Tina > >St. Joseph's Histology >St. Paul, MN > > > >The information included in this e-mail message, including any >attachments, is intended only for the person or organization to which it >is addressed. This e-mail message may contain information that is >privileged or confidential. If you receive this e-mail message and are >not the intended recipient or responsible for delivering the message to >the intended recipient, you may not use, disseminate, distribute or copy >the information included in this e-mail and any attachments. If you >received this e-mail message by mistake, please reply by e-mail and >destroy all copies of this message and any attachments. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HornHV <@t> archildrens.org Wed Sep 21 15:24:24 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Wed Sep 21 15:24:14 2005 Subject: [Histonet] plants in histology lab Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDE32@EMAIL.archildrens.org> We do have plants in our lab. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mrosla, Tina M Sent: Wednesday, September 21, 2005 12:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] plants in histology lab We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From ynwang <@t> u.washington.edu Wed Sep 21 15:38:40 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Wed Sep 21 15:38:55 2005 Subject: [Histonet] ultralow freezer Message-ID: Hi everyone, Can people tell me if they have a chest or upright ultra low freezer and which they prefer. A colleague is looking for a small ultra low. Also, any particular recommendations would be greatly appreciated. Thanks Yak-Nam From Jackie.O'Connor <@t> abbott.com Wed Sep 21 17:13:57 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Sep 21 17:14:28 2005 Subject: [Histonet] ultralow freezer Message-ID: I absolutely detest upright ultra lows. When you open them, all the cold air falls out,condensed ice on the jamb falls on the floor, and you don't have much time to search for what is buried under someone else's Popsicles before it starts to go out of spec. I much prefer the chest type (coffin) freezers. The cold air stays in, and they're easier to search. I currently have two uprights and one chest freezer - one of the uprights is trying to die - I can hardly wait so I can get another coffin. The one drawback is space. The coffins take up a lot of room. Jackie O' "Y. Wang" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/21/2005 03:38 PM To: Histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] ultralow freezer Hi everyone, Can people tell me if they have a chest or upright ultra low freezer and which they prefer. A colleague is looking for a small ultra low. Also, any particular recommendations would be greatly appreciated. Thanks Yak-Nam _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Raoul.Regnault <@t> interiorhealth.ca Wed Sep 21 17:20:14 2005 From: Raoul.Regnault <@t> interiorhealth.ca (Regnault, Raoul) Date: Wed Sep 21 17:20:41 2005 Subject: [Histonet] CJD protocols anyone? Message-ID: Hello Julia Your situation sounds similar to ours here. Following is the information I discovered for us in Canada. Raoul Regnault Anatomic Pathology Kootenay Boundary Regional Hospital Trail B.C. 250-368-3311 ext 2263 fax 364-3421 raoul.regnault@interiorhealth.ca ************************ CJD Information - Canada - September 2005 Creutzfeldt-Jakob Disease Surveillance System (CJD-SS) 1-888-489-2999 http://www.phac-aspc.gc.ca/hcai-iamss/cjd-mcj/index.html Elina Olsen -coordinator - 1-613-946-9864 The toll free number is manned, responsive and is the preferred access phone number.(see above) CJD-SS provides approved shipping containers, no cost. The Public Health Agency of Canada provides a manual: Volume: 28S5,November 2002, Infection Control Guidelines, Classic Creutzfeldt-Jakob Disease in Canada. The URL is as follows; http://www.phac-aspc.gc.ca/publicat/ccdr-rmtc/02vol28/28s5/index.html A printed copy of this manual is available in Anatomic Pathology, KBRH (09/05) Prion Lab (? National) Ottawa Civic Hospital 1653 Carling Avenue Ottawa, Ontario K1Y 4E9 Att: Laboratory Medicine, Dr. G. Jansen Dr. Gerard Jansen Neuropathologist 1-613-798-5555 ext 19847 Pager 1-613-715-7023 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Celebre Julia Sent: Wednesday, September 21, 2005 9:37 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] CJD protocols anyone? Hi there Histoland! Our department has been hit by several 'query' CJD surgical cases lately, since we rarely handled any questionable cases in the past we are trying to prepare an updated protocol that will include the handling of these specimens from begining to end. I would appreciate hearing from anyone who deals with CJD cases from safety protocols, protocols on deactivation, processing, cutting and staining, everything and anything dealing with CJD. Thanks Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca The Cornerstone of Care Campaign for Hamilton Health Sciences needs your support. Please, visit hamiltonhealthsciences.ca today and help make something great even greater. This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Melissa.Gonzalez <@t> cellgenesys.com Wed Sep 21 18:28:53 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Wed Sep 21 18:29:17 2005 Subject: [Histonet] Fluorescent Illumination Systems Message-ID: Hello all, I am currently in the market to upgrade our existing HB0 100 lamp house, which is very old, to several of new systems. Either the newest version of the HB0 100 from Zeiss, a mercury halide system called X-Cite from EXFO, or a xenon based burner called Lambda LS from Sutter Instruments. They are all comparable in price, but my main concern is increasing the brightness of my signal and evenly illuminating the field of view. I mostly look at DAPI, eGFP, FITC, and Alexa 594, which from spectral outputs, each one of these systems appears to have a different weakness in one of the above. On a plus, the Xcite rates their bulbs at 1500 hours, and the Lambda at 500-1000hrs. Does anyone out there have any input/comments/experience with these systems? (This would be hooked up to a Zeiss Axioplan) Thanks a lot, Melissa From jwatson <@t> gnf.org Wed Sep 21 18:45:50 2005 From: jwatson <@t> gnf.org (James Watson) Date: Wed Sep 21 18:46:11 2005 Subject: [Histonet] Citraconic anhydride Message-ID: I was contacted today by the sales person that told me that Biocare sold a Citraconic anhydride antigen retrieval solution and was informed that she was misinformed about the make up of the new antigen retrieval solution. She apologized for the misinformation. And I apologize for repeating it. For my experience with using 0.05% Citraconic per the Namimatsu, Ghazizadeh, and Sugisaki paper J Histochem Cytochem 53:3-33,2005 paper: I have found in preliminary testing that it works well on many antibodies that I normally use citrate buffer pH 6.0 or tris/EDTA pH 9.0 , but on some antibodies stronger signal can be seen using other retrieval solutions. The testing continues. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Tuesday, September 20, 2005 8:43 AM To: Histonet Cc: ihcrg@neo.agsci.colostate.edu Subject: [Histonet] Citraconic anhydride My apologies in advance for those of you on both listserves... In one of my IHC workshops at the NSH, the subject of citraconic anhydride came up. Apparently a couple of people have tried it but didn't get the dramatic effects as seen in the JHC paper. Somebody mentioned one company has this commercially available, and I'm interested if this is true, which company, and any experiences using this chemical, commercial or otherwise. 'Tis a nasty chemical, and one I may well be staying away from. Best wishes, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From AnthonyH <@t> chw.edu.au Wed Sep 21 18:56:34 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Wed Sep 21 18:57:18 2005 Subject: [Histonet] plants in histology lab Message-ID: The plants I would suggest are: Peace Lily (Spathiphyllum) Gerbera (Gerbera jamesonii) Chrysanthemum (Chrysanthemum morifolium) - good for removing formaldehyde, benzene and ammonia from the air. Boston Fern (Nephrolepsis exaltata "Bostoniensis") - suggested to be the best for removing formaldehyde. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mrosla, Tina M Sent: Thursday, 22 September 2005 3:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] plants in histology lab We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From Kemlo.Rogerson <@t> elht.nhs.uk Thu Sep 22 02:20:59 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Thu Sep 22 02:21:23 2005 Subject: [Histonet] plants in histology lab Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E699037@elht-exch1.xelht.nhs.uk> Sapium Sebiferum 'Chinese Tallow Tree)? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: 21 September 2005 20:17 To: tmmrosla@healtheast.org; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] plants in histology lab I've had a bonsai tree in my lab for the past 5 years and it's doing great. Hard to say if the fumes contribute to it's contorted appearance. Fred >>> "Mrosla, Tina M" 09/21/05 01:45PM >>> We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Thu Sep 22 02:26:54 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Thu Sep 22 02:27:10 2005 Subject: [Histonet] plants in histology lab Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E69903A@elht-exch1.xelht.nhs.uk> Best use them like canaries they used down coal mines; if the plants die, get out. If they live keep a watchful eye on them; feng shui uses plants in strategic areas of the room, near doors I think. Get the Company to buy them as a safety device and for stress relief. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mrosla, Tina M Sent: 21 September 2005 18:45 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] plants in histology lab We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Thu Sep 22 02:33:40 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Thu Sep 22 02:36:27 2005 Subject: [Histonet] RE: CJD Message-ID: Try CJD Surveillance Unit Old Pharmacy Building Western General Hospital Crewe Road Edinburgh EH4 2XU Scotland Tel: 0131 332 2117 Fax: 0131 343 1404 Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From carl.hobbs <@t> kcl.ac.uk Thu Sep 22 03:03:57 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Thu Sep 22 03:05:02 2005 Subject: [Histonet] re: Fluorescent Illumination Systems Message-ID: <001201c5bf4c$2ed9c2d0$112b5c9f@Carlos> I recently upgraded my three Axioskops/verts from 50W mercury to EXfo's XCite 100W lamps. I have no regrets and highly recommend them: we use DAPI, Hoechst, Alexas 488 and 594 routinely. To me, the 100W mercury vapour lamps still have the"edge" over the Xcite lamps( I have one as well) in brightness, tho. But, given the ease of use, the longevity of the lamps and the evenness of the illumination field I would recommend the Xcite lamps. NB: they create fan- noise, tho! Particularly the older ones; their new models are much quieter Test one out, anyway. From TJJ <@t> Stowers-Institute.org Thu Sep 22 07:14:46 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Thu Sep 22 07:15:05 2005 Subject: [Histonet] RE: Brain tissue frozen section Message-ID: >< Then we put the tissue (rat or mouse brain) into 30% sucrose in 0.1 M phosphate buffer (PB) and leave it for at least overnight (you don't say how long you leave it) or until the tissue floats. >< That should read until the tissue sinks. :) It's much easier to section this material if you have a cryostat that has a temperature control on the specimen holder. I will echo the suggestion that you try different freezing media, although we have never had any trouble with the OCT compound. We have only done a couple rodent brain frozen sections though. If your section is laying flat on your blade/blade holder before picking up, then part of the problem is in your mounting technique. I recommend starting the pickup at one end of your tissue (at the edge closest to you towards the bottom of the slide) and then learn to move the slide slightly towards you as you lower it down to the blade. If you just plop the slide down flat on your section, you will introduce wrinkles and bubbles in your section. Admittedly, I've never totally perfected whole brain sections and am awed by those publications that show flawless sections (but wonder how many it took to get that look!). Good luck and let us know what works for you! Teri Johnson Stowers Institute for Medical Research Kansas City, MO From gcallis <@t> montana.edu Thu Sep 22 10:54:17 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Sep 22 10:54:16 2005 Subject: [Histonet] Mitochondria fluorescent marker Message-ID: <6.0.0.22.1.20050922095329.01b4b7f8@gemini.msu.montana.edu> Check with Molecular Probes on their website. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From ree3 <@t> leicester.ac.uk Thu Sep 22 11:12:00 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Thu Sep 22 11:12:26 2005 Subject: [Histonet] haemoxygenase I or II Message-ID: Wanted antibodies to the above that work on mouse paraffin processed tissues...thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K..... From dpahisto <@t> yahoo.com Thu Sep 22 11:50:42 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Thu Sep 22 11:51:00 2005 Subject: [Histonet] Plants in lab Message-ID: <20050922165043.65335.qmail@web33413.mail.mud.yahoo.com> We have several plants in our lab. African Violets seem to do extremely well and bloom all year round. It adds some nice color to our environment. Cindy DuBois Delta Pathology Assoc. Stockton CA --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From Terry.Marshall <@t> rothgen.nhs.uk Thu Sep 22 11:57:39 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Thu Sep 22 11:57:32 2005 Subject: [Histonet] Plants in lab Message-ID: I'm all for plants in he lab. (or anywhere). A Norfolk pine seems a tad extravagant. I've seen them about 20 metres tall! Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Cindy DuBois [mailto:dpahisto@yahoo.com] Sent: 22 September 2005 17:51 To: tmmrosla@healtheast.org Cc: Histonet Subject: [Histonet] Plants in lab We have several plants in our lab. African Violets seem to do extremely well and bloom all year round. It adds some nice color to our environment. Cindy DuBois Delta Pathology Assoc. Stockton CA --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Thu Sep 22 12:12:01 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Thu Sep 22 12:12:21 2005 Subject: [Histonet] anti-Hu antibody Message-ID: Hello, Our neuropathologist would like to get an anti-Hu stain done on some autopsy material. Are there any reference labs out there that perform this immunohistochemical stain? Thanks, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From BlazekL <@t> childrensdayton.org Thu Sep 22 12:32:20 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Thu Sep 22 12:33:07 2005 Subject: [Histonet] Plants in lab Message-ID: The Norfolk pine started out as a seed 10 years ago. It is now around a meter. When there are no windows I guess the growth rate is rather slow. Or we need to find better fertilizer. Linda >>> "Marshall Terry Dr, Consultant Histopathologist" 09/22/2005 12:57 PM >>> I'm all for plants in he lab. (or anywhere). A Norfolk pine seems a tad extravagant. I've seen them about 20 metres tall! Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Cindy DuBois [mailto:dpahisto@yahoo.com] Sent: 22 September 2005 17:51 To: tmmrosla@healtheast.org Cc: Histonet Subject: [Histonet] Plants in lab We have several plants in our lab. African Violets seem to do extremely well and bloom all year round. It adds some nice color to our environment. Cindy DuBois Delta Pathology Assoc. Stockton CA --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From 1dpeterson <@t> meriter.com Thu Sep 22 12:41:12 2005 From: 1dpeterson <@t> meriter.com (Peterson, Dan) Date: Thu Sep 22 12:41:31 2005 Subject: [Histonet] RE: Histonet Digest, Vol 22, Issue 27 Message-ID: <328CBAE62F31C642B422970E879DFADC01A7FFD3@pcwex01> We've got about a dozen different types of plants of various shapes and sizes, even a rubber tree. They thrive very well on the fumes. That and the fish water from our 50 gal. aquarium................ Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories (608)-267-6557 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Thursday, September 22, 2005 12:03 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 22, Issue 27 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: CJD (Malam Jacqueline) 2. re: Fluorescent Illumination Systems (Carl Hobbs) 3. RE: Brain tissue frozen section (Johnson, Teri) 4. Mitochondria fluorescent marker (Gayle Callis) 5. haemoxygenase I or II (Edwards, R.E.) 6. Plants in lab (Cindy DuBois) 7. RE: Plants in lab (Marshall Terry Dr, Consultant Histopathologist) ---------------------------------------------------------------------- Message: 1 Date: Thu, 22 Sep 2005 08:33:40 +0100 From: Malam Jacqueline Subject: [Histonet] RE: CJD To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Try CJD Surveillance Unit Old Pharmacy Building Western General Hospital Crewe Road Edinburgh EH4 2XU Scotland Tel: 0131 332 2117 Fax: 0131 343 1404 Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 2 Date: Thu, 22 Sep 2005 09:03:57 +0100 From: "Carl Hobbs" Subject: [Histonet] re: Fluorescent Illumination Systems To: Message-ID: <001201c5bf4c$2ed9c2d0$112b5c9f@Carlos> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original I recently upgraded my three Axioskops/verts from 50W mercury to EXfo's XCite 100W lamps. I have no regrets and highly recommend them: we use DAPI, Hoechst, Alexas 488 and 594 routinely. To me, the 100W mercury vapour lamps still have the"edge" over the Xcite lamps( I have one as well) in brightness, tho. But, given the ease of use, the longevity of the lamps and the evenness of the illumination field I would recommend the Xcite lamps. NB: they create fan- noise, tho! Particularly the older ones; their new models are much quieter Test one out, anyway. ------------------------------ Message: 3 Date: Thu, 22 Sep 2005 07:14:46 -0500 From: "Johnson, Teri" Subject: [Histonet] RE: Brain tissue frozen section To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" >< Then we put the tissue (rat or mouse brain) into 30% sucrose in 0.1 M phosphate buffer (PB) and leave it for at least overnight (you don't say how long you leave it) or until the tissue floats. >< That should read until the tissue sinks. :) It's much easier to section this material if you have a cryostat that has a temperature control on the specimen holder. I will echo the suggestion that you try different freezing media, although we have never had any trouble with the OCT compound. We have only done a couple rodent brain frozen sections though. If your section is laying flat on your blade/blade holder before picking up, then part of the problem is in your mounting technique. I recommend starting the pickup at one end of your tissue (at the edge closest to you towards the bottom of the slide) and then learn to move the slide slightly towards you as you lower it down to the blade. If you just plop the slide down flat on your section, you will introduce wrinkles and bubbles in your section. Admittedly, I've never totally perfected whole brain sections and am awed by those publications that show flawless sections (but wonder how many it took to get that look!). Good luck and let us know what works for you! Teri Johnson Stowers Institute for Medical Research Kansas City, MO ------------------------------ Message: 4 Date: Thu, 22 Sep 2005 09:54:17 -0600 From: Gayle Callis Subject: [Histonet] Mitochondria fluorescent marker To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050922095329.01b4b7f8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Check with Molecular Probes on their website. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 5 Date: Thu, 22 Sep 2005 17:12:00 +0100 From: "Edwards, R.E." Subject: [Histonet] haemoxygenase I or II To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Wanted antibodies to the above that work on mouse paraffin processed tissues...thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K..... ------------------------------ Message: 6 Date: Thu, 22 Sep 2005 09:50:42 -0700 (PDT) From: Cindy DuBois Subject: [Histonet] Plants in lab To: tmmrosla@healtheast.org Cc: Histonet Message-ID: <20050922165043.65335.qmail@web33413.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 We have several plants in our lab. African Violets seem to do extremely well and bloom all year round. It adds some nice color to our environment. Cindy DuBois Delta Pathology Assoc. Stockton CA --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. ------------------------------ Message: 7 Date: Thu, 22 Sep 2005 17:57:39 +0100 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Plants in lab To: "Cindy DuBois" , Cc: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" I'm all for plants in he lab. (or anywhere). A Norfolk pine seems a tad extravagant. I've seen them about 20 metres tall! Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Cindy DuBois [mailto:dpahisto@yahoo.com] Sent: 22 September 2005 17:51 To: tmmrosla@healtheast.org Cc: Histonet Subject: [Histonet] Plants in lab We have several plants in our lab. African Violets seem to do extremely well and bloom all year round. It adds some nice color to our environment. Cindy DuBois Delta Pathology Assoc. Stockton CA --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 27 **************************************** From JEllin <@t> yumaregional.org Thu Sep 22 13:15:37 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Thu Sep 22 13:17:07 2005 Subject: [Histonet] Microwave PROCCESSING Message-ID: Nice to see that everyone made it back from NSH and are doing well. I have a nice question that might start some serious discussion. Microwave proccessing!!!! My question is how many people out there are doing it and what issues did you have to work through to apply this technology to everyday use??? How are you handling the testing of your immuno's since you have to use tissue that has been run by the microwave proccessor??? How is the transcription being done??? What time are the pathologist coming in and how LATE are they staying?? Are you proccessing all tissue type or only small biopies, breast biopsies, etc.?? What road block have you come up against??? It this the rule to proccess all tissue or is this the exception?? How are you handling reference lab consultation with your tissue that is proccessed on the Microwave and sending it to a lab for instance AFIP for consult and they only use the conventional proccessing?? Is anyone out there doing both and how do they keep track of the different types of tissue that have been proccessed within the different proccessor, if you are indeed using both. What additional costs did you do you have, with there new product from the proccessor?? Our pathologist came back and are now foaming at the mouth for this technology. We are not against this, but there are avenues that need to be addressed, especially with IHC, FISH, and overall workflow. Any help would greatly be appreciated and needed at this time. Your in Formalin Jesus Ellin Yuma Regional Medical Center From jgemmanual <@t> wlgore.com Thu Sep 22 13:33:58 2005 From: jgemmanual <@t> wlgore.com (Jeannie G Emmanuel) Date: Thu Sep 22 13:34:23 2005 Subject: [Histonet] Leica or Sakura? Message-ID: Hi all! I am looking to purchase a tissue processor for my plastics histo lab. My processing schedules are usually more than 24 hrs long, sometimes much longer. Does anyone know which is a better, friendlier, simpler processor out there. Currently, I have a Leica on demo and I can't just tell the machine to get it done by a certain time of the day (be it 2 day 3 day or 5 days later) and expect the machine to know to delay the process at station 1. The support person at Leica told me to go to Programs and select the day I want the blocks to come out and then it will know to go on delay mode. But when I tried to do that it asked for the supervisor password. We don't have supervisors so no job functions are limited to supervisors only besides my lab is a one person lab. One might argue that once I purchased it I will authorize my own password and that step becomes a routine. But a little bit of history here.... Currently we have an old Sakura tissue tek and it is definitely more intelligent than the new Leica ASP 300, I tell it when to spit out the blocks and it knows to sit in station 1 for however long. Plus all my schedules are always more than 24 hrs, so using the Leica ASP 300 will always involve that extra two-three step programming, Plus it is not easy to get Leica personnel to return your phone calls! So I am wondering if Sakura processor is more user friendly. Any thoughts from you will be appreciated...thanks! From laurie.reilly <@t> jcu.edu.au Thu Sep 22 13:33:59 2005 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Thu Sep 22 13:34:29 2005 Subject: [Histonet] Plants in lab In-Reply-To: Message-ID: <5.1.0.14.0.20050923043138.00c3da90@mail.jcu.edu.au> Why not a Logwood tree, Haematoxylum campechianum, they make great bonsai specimens? Regards, Laurie. At 05:57 PM 22/09/2005 +0100, Marshall Terry Dr, Consultant Histopathologist wrote: >I'm all for plants in he lab. (or anywhere). > >A Norfolk pine seems a tad extravagant. I've seen them about 20 metres tall! > >Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path > Consultant Pathologist > Rotherham General Hospital > South Yorkshire > England > terry.marshall@rothgen.nhs.uk > >-----Original Message----- >From: Cindy DuBois [mailto:dpahisto@yahoo.com] >Sent: 22 September 2005 17:51 >To: tmmrosla@healtheast.org >Cc: Histonet >Subject: [Histonet] Plants in lab > > >We have several plants in our lab. African Violets seem to do extremely >well and bloom all year round. It adds some nice color to our environment. > >Cindy DuBois >Delta Pathology Assoc. >Stockton CA > > >--------------------------------- >Yahoo! for Good > Click here to donate to the Hurricane Katrina relief effort. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From dsnider <@t> shrinenet.org Thu Sep 22 13:54:52 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Thu Sep 22 13:55:20 2005 Subject: [Histonet] Embedding centers Message-ID: <84BE46B37B314D409C5A17B7BAB022D622500A@IDC-EX-VS01.shriners.cc> Hi everyone! I once again am in need of your knowledge and guidance! My existing embedding center is on the fritz. We are looking into purchasing a new one. Which brands do you recommend? Why? I am a low volume research lab, working with clinically engineered skin and human on mouse grafts. I looked at the archives but not much was there! Thanks in advance, Deanna Snider HT ASCP Shriners Hospital for Children Research Dept. Cincinnati, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From JSCHUMA1 <@t> Fairview.org Thu Sep 22 14:06:55 2005 From: JSCHUMA1 <@t> Fairview.org (Schumacher, Jennifer J) Date: Thu Sep 22 14:07:16 2005 Subject: [Histonet] Leica or Sakura? Message-ID: We have the Leica and get superb customer service and prompt attention. We are in Minnesota. However, whoever set up your demo should have "turned off" the supervisor capability. Then, you would have access to all of the necessary functions. It is easy to do, but I don't think you can do it in operator mode. We have old Sakura VIP processors as well as the Leica, and I think the Leica has been a huge improvement, especially when needing to change the end day/time. Ask Leica to turn off the password requirement, and I think you will be pleasantly surprised by it's ease of use. Just my two cents. Jennifer -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jeannie G Emmanuel Sent: Thursday, September 22, 2005 1:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Leica or Sakura? Hi all! I am looking to purchase a tissue processor for my plastics histo lab. My processing schedules are usually more than 24 hrs long, sometimes much longer. Does anyone know which is a better, friendlier, simpler processor out there. Currently, I have a Leica on demo and I can't just tell the machine to get it done by a certain time of the day (be it 2 day 3 day or 5 days later) and expect the machine to know to delay the process at station 1. The support person at Leica told me to go to Programs and select the day I want the blocks to come out and then it will know to go on delay mode. But when I tried to do that it asked for the supervisor password. We don't have supervisors so no job functions are limited to supervisors only besides my lab is a one person lab. One might argue that once I purchased it I will authorize my own password and that step becomes a routine. But a little bit of history here.... Currently we have an old Sakura tissue tek and it is definitely more intelligent than the new Leica ASP 300, I tell it when to spit out the blocks and it knows to sit in station 1 for however long. Plus all my schedules are always more than 24 hrs, so using the Leica ASP 300 will always involve that extra two-three step programming, Plus it is not easy to get Leica personnel to return your phone calls! So I am wondering if Sakura processor is more user friendly. Any thoughts from you will be appreciated...thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Thu Sep 22 14:11:39 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Thu Sep 22 14:12:03 2005 Subject: [Histonet] Embedding centers Message-ID: I love my Sakura! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Snider, Deanna [mailto:dsnider@shrinenet.org] Sent: Thursday, September 22, 2005 11:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Embedding centers Hi everyone! I once again am in need of your knowledge and guidance! My existing embedding center is on the fritz. We are looking into purchasing a new one. Which brands do you recommend? Why? I am a low volume research lab, working with clinically engineered skin and human on mouse grafts. I looked at the archives but not much was there! Thanks in advance, Deanna Snider HT ASCP Shriners Hospital for Children Research Dept. Cincinnati, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Thu Sep 22 14:30:51 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Thu Sep 22 14:31:15 2005 Subject: [Histonet] Integrin beta1 / CD29 -- cell surface or not?? Message-ID: <20050922193051.62672.qmail@web31707.mail.mud.yahoo.com> Hello all, Does anyone have experience with the antigen? I am using a monoclonal rat anti-mouse integrin beta1 (clone 265917) from R&D systems with a goat anti-rat (Alexa-594 conj.) secondary -- DAPI nuclear staining. I am staining mouse aortic sinuses with atherosclerotic lesions. Today was my first run and I expected to get nice cell membrane staining red. However, I am getting little 'cell' membrane and more nuclear membrane (?) staining. How can this be? The specificity should be to "detect mouse Integrin beta1" -- CD29 (cell surface??). My negatives are mostly clear except for some non-specific binding (Ab concentration to high) along the smooth muscle cells -- no nuclear staining seen in negative tells me my positive is true positive. I need a really good cell surface marker so that I can run double immunos to determine if there is co-localization of my antigen on the cell surface. If anyone has any suggestions, answers or questions for that matter ... anything would be very helpful. Also, if you have run immunofluorescence with the antibody or other well-definied cell surface markers, photos of what I am supposed to be seeing would also be very helpful. Thank you. Gustave Hebert Scientist II Wyeth Research Cambridge MA email photos: emerald_lake77@yahoo.com --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From Diane.Gladney <@t> se.amedd.army.mil Thu Sep 22 14:41:58 2005 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Thu Sep 22 14:39:42 2005 Subject: [Histonet] Leica or Sakura? Message-ID: <4D55B2E997EFAE4DA6081DDE100B8302822550@amedmlsermc133.amed.ds.army.mil> Jeannie, I have a new Leica ASP 300 and love it. You can set the processor to the supervisor mode (you don't have to enter a supervisor password if you leave it in this mode all the time) therefore you will not need a password. The supervisor symbol is the "red shirt" symbol (found in the upper right corner) or on the "Smart Screen" on the right bottom next to the "I" symbol for information. The "operator mode" is a "yellow shirt". The password is only needed when the processor is in the operator mode (operator is limited to access to certain functions). I run mine in the supervisor mode all the time. I have different programs set to end on different days....overnight, weekend, 3 day weekend, etc. You can select which station that you want the specimens to be held in for the delay, if you wish. This processor is very versatile and I find it easy to use. Did they not give you an Owner's Manual to help you? This processor is one of the easiest to use once the system is set up for your processing schedules and the reagent management system is great. It is a shame that your Leica Sales Representative hasn't been of more help. I have had great support from my representative and from the office in Chicago during my "learning phase". The contact that I have at the Chicago Office for help is: Mari Ann Mailhiot, 800-248-0123 ext. 7267. She has been most helpful. Let me know if I can be of further help. Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Street P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jeannie G Emmanuel Sent: Thursday, September 22, 2005 2:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Leica or Sakura? Hi all! I am looking to purchase a tissue processor for my plastics histo lab. My processing schedules are usually more than 24 hrs long, sometimes much longer. Does anyone know which is a better, friendlier, simpler processor out there. Currently, I have a Leica on demo and I can't just tell the machine to get it done by a certain time of the day (be it 2 day 3 day or 5 days later) and expect the machine to know to delay the process at station 1. The support person at Leica told me to go to Programs and select the day I want the blocks to come out and then it will know to go on delay mode. But when I tried to do that it asked for the supervisor password. We don't have supervisors so no job functions are limited to supervisors only besides my lab is a one person lab. One might argue that once I purchased it I will authorize my own password and that step becomes a routine. But a little bit of history here.... Currently we have an old Sakura tissue tek and it is definitely more intelligent than the new Leica ASP 300, I tell it when to spit out the blocks and it knows to sit in station 1 for however long. Plus all my schedules are always more than 24 hrs, so using the Leica ASP 300 will always involve that extra two-three step programming, Plus it is not easy to get Leica personnel to return your phone calls! So I am wondering if Sakura processor is more user friendly. Any thoughts from you will be appreciated...thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Thu Sep 22 14:57:38 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Thu Sep 22 14:58:08 2005 Subject: [Histonet] Kim Pittelko? Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D151@usca0082k08.labvision.apogent.com> Anyone know the whereabouts of Kim Pittelko, formerly of Berkeley, CA? Tim Morken Lab Vision - Neomarkers From kbradshaw <@t> lcpath.com Thu Sep 22 15:51:47 2005 From: kbradshaw <@t> lcpath.com (Kari Bradshaw) Date: Thu Sep 22 15:52:17 2005 Subject: [Histonet] Kim Pittelko? In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D151@usca0082k08.labvision.apogent.com> Message-ID: <200509222051.j8MKppA07595@plus34.host4u.net> Tim, If you call me I will give you her contact information. Kari L. Bradshaw Laboratory Manager Lower Columbia Pathologists 1217 14th Ave Longview, WA 98632 360-425-5620 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Thursday, September 22, 2005 12:58 PM To: Histonet Subject: [Histonet] Kim Pittelko? Anyone know the whereabouts of Kim Pittelko, formerly of Berkeley, CA? Tim Morken Lab Vision - Neomarkers _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ynwang <@t> u.washington.edu Thu Sep 22 18:51:38 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Thu Sep 22 18:51:57 2005 Subject: [Histonet] ultra low freezer Message-ID: Thanks for advice and info on ultra low. I have passed on messages to my colleague. I think they are going to go for a small revco chest freezer. Yak-Nam From djohnson14 <@t> hotmail.com Thu Sep 22 19:39:26 2005 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Thu Sep 22 19:39:44 2005 Subject: [Histonet] Sakura-esque slide racks In-Reply-To: <0C44F1AAEE47D54DA4210A60AB206F5E0445CBFF@SKMCEMAIL.skmc.gov.ae> Message-ID: Mercedes Medical Phone #800-331-2716 We have a generic rack that suits your request Part # CON 1000- $100 per 10 racks Let me know if we can help >From: "Anne Van Binsbergen" >To: "Breeden, Sara" >, >Subject: RE: [Histonet] Sakura-esque slide racks >Date: Tue, 20 Sep 2005 07:52:27 +0400 > >Try the Sakura website - they have them >Annieinarabia > >-----Original Message----- >From: Breeden, Sara [mailto:sbreeden@nmda.nmsu.edu] >Sent: Tuesday, September 20, 2005 1:03 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Sakura-esque slide racks > >I'm on the hunt for those Sakura 20-slide racks for tape coverslippers; >handles, too. I'm sure this has been covered on 'net before, but could >someone direct me? Vendors are welcome to contact me, but be it known >that I'm only collecting information at this point... > >Sally Breeden, HT(ASCP) >New Mexico Department of Agriculture >Veterinary Diagnostic Services >POBox 4700 >Albuquerque, NM 87196 >(505)841-2576 >(505)841-2518 FAX > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Eric <@t> ategra.com Thu Sep 22 20:58:10 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Thu Sep 22 20:49:56 2005 Subject: [Histonet] Histo Tech Supervisors, and Bench techs Needed for Immediate openings Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. (I have a few travel/temp HistoTech positions as well - see below). Most of my Histo Techs positions are clinical, although I do have a few Research Histo Tech positions as well. Here are some of my Hottest jobs: 1. Central Florida(Ocala/Gainsville) Openings for a HistoTech(Bench) No weekends, No call, and Top Dollar. 2. Ohio(Moraine Area) HistoTech Manager No Weekends, No call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 3. Colorado (Greater Denver area) Histotech Supervisor Opening for Several Bench Positions(Night Shift) No weekends, No call, Top Dollar 4. New Hampshire Openings for several Histotechs(Bench) No Weekends, No Call, Top Dollar, Excellent Benefits if you are interested, please call me today at 1-800-466-9919 ext 223 5. Minnesota Opening for HistoTech(Bench) No weekends, No Call, Top Dollar 6. New Jersey(Southern) Openings for HistoTech(Bench) No Weekends, No Call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 7.Oregon(Portland area) Openings for Histotech(Bench) No weekends, No call, Top Dollar 8. California( Southern) HistoTech Supervisor(2nd shift) Openings for Histotechs(Bench,2nd shift) Great Location, No weekends, No call, Top Dollar 9. California(Southern) Openings for HistoTechs(Bench) Great Location, No weekends, No call, Top Dollar 10. Pennsylvania HistoTech opening(Bench) No weekends, No call if you are interested, please call me today at 1-800-466-9919 ext 223 11. West Virginia (west of Hagerstown,MD) Opening for Histotech(Bench) No weekends, No call, Top Dollar 12. Illinois(Chicagoland) Openings for Histotechs(Bench) Opening for Pathologist Assistant No weekends, No Call, Top Dollar 13. Illinois(South of Chicago) Opening for Histotech(Bench) No weekends, No call, Top Dollar 14. Washington State(Eastern) Opening for HistoTech(Bench) No weekends, No call 15. Delaware Opening for MOHS Histotech(Bench) No weekends, No call, Free training,Brand new Lab. 16. Moussouri Histotech Supervisor No weekends, No call 17.Kentucky No Weekends,Very light Call, Great Benefits The clients are currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 From dbpiontek <@t> hotmail.com Thu Sep 22 21:40:08 2005 From: dbpiontek <@t> hotmail.com (Denise Piontek) Date: Thu Sep 22 21:40:42 2005 Subject: [Histonet] Job Opportunity in MA Message-ID: Contract Research Organization specializing in biocompatibility and toxicity studies for the medical device and pharmaceutical industries is in need of a full-time, experienced histology technician or histotechnologist due to major expansion. The person, as a team player, will work with other histology technicians, veterinarian, pathologist, Study Directors, and animal technicians to produce quality work. Responsibilities will include: * Trimming / grossing of tissues and organs harvested at necropsy * Embedding and processing of tissues * Microtomy * Routine H&E staining, special stains as required and special techniques as attained * Maintain and/or create GLP documentation as necessary, including relative SOPs and histology records * ISO regulated facility * Assist in necropsy procedures as necessary * Maintain wet tissue archive * Data evaluations of various project aspects * Great opportunity to increase or use anatomy skills Experience and an AS degree are required. Research experience, certification by the ASCP (or eligible), and/or experience with animal tissues are strongly preferred. Fully automated laboratory, experience with equipment a plus. Willingness to learn, attention to detail, proficiency with computers, good organizational skills, and well-developed time management skills are also desired. Salary is commiserate with experience. Competitive benefits. Position is M-F 7AM to 4 PM or 8AM to 5 PM. We are located just outside of Boston, MA. Please submit resumes to: Human Resources Toxikon Corporation 15 Wiggins Ave. Bedford, MA 01730 Email your resume to: [1]hr@toxikon.com Visit us on the web at [2]www.toxikon.com _________________________________________________________________ [3]With MSN Spaces email straight to your blog. Upload jokes, photos and more. It's free! References 1. http://by24fd.bay24.hotmail.msn.com/cgi-bin/compose?curmbox=F000000001&a=e752b25041c2d8a23271caeb4e225dcb56d27008f1b7b3264bb774f89eacbb1d&mailto=1&to=hr@toxikon.com&msg=MSG1125799332.18&start=4868040&len=5464&src=&type=x 2. http://www.toxikon.com/ 3. http://g.msn.com/8HMAENUS/2746??PS=47575 From tyler-wellington <@t> northwestern.edu Fri Sep 23 00:20:46 2005 From: tyler-wellington <@t> northwestern.edu (Tyler W.) Date: Fri Sep 23 00:21:02 2005 Subject: [Histonet] sectioning problems Message-ID: <20050923052046.8C2919E851@merle.it.northwestern.edu> I recently have been having problems pulling ribbons from my microtome and was wondering if anyone has advice. 1.) I was having problems pulling ribbons from newly embeded blocks, but ones that had been embeded previously were very easy to pull ribbons from. The new ones would simply curl on the blade and wouldn't even grab on to the adjascent sections. 2.) I changed the paraffin in the embedding station because I noticed all the temeratures were set to 64 C when the melting point for the Surgipath paraffin formula 'R' was 56-67C. Could this have been a problem? 3.) I switched to an old lot of Fisher embeding medium and now all the section come off, but on occasion when the chuck on the mictomome comes back up it seems to (via static electricity?) pull the ribbon from the microtome blade and interupt the ribbon. 4.) Also, visually the Fisher paraffin blocks look almost transparent whereas the Surgipath blocks seem opaque and white. I'm relatively new to the histology game and I realize that there is both art and science involved, but this is clearly unacceptable behavior from the paraffin, something is surely wrong. I'm sure you are all experienced and maybe you have advice? If you do I would apreciate it so much. The first and best advice get their choice of FTD flowers and/ or homemade brownies. Thank you--Tyler Wellington From Susan.Ferrigon <@t> sanofi-aventis.com Fri Sep 23 05:27:59 2005 From: Susan.Ferrigon <@t> sanofi-aventis.com (Susan.Ferrigon@sanofi-aventis.com) Date: Fri Sep 23 05:28:19 2005 Subject: [Histonet] Antigen retreival on mouse GFAP. Message-ID: Hi Does anyone have any experience with using trypsin as the method of antigen retreival when staining GFAP on mouse brains??? Any advice on concentration, temperature and incubation time would be appreciated. Thanks Susan. ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From jennifer.l.hofecker <@t> Vanderbilt.Edu Fri Sep 23 08:23:37 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Fri Sep 23 08:27:36 2005 Subject: [Histonet] removing silver from counters, etc. Message-ID: <898D946569A27444B65667A49C0740520754B1@mailbe06.mc.vanderbilt.edu> Happy Friday everyone. Can somebody remind me what solutions will remove silver nitrate from cabinets, floors, counters, etc.? Bleach just changes these spots to different shades of brown. I remember using solution from hair perms in the past but I am sure that there are other reliable ways. Thanks for any advice. Our lab is starting to resemble a dalmation! You know how silver solution likes to migrate from the coplin jars! Thoughts and prayers go out to Gulf Coast residents. Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From cgfields <@t> lexhealth.org Fri Sep 23 08:36:07 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 23 08:34:37 2005 Subject: [Histonet] removing silver from counters, etc. Message-ID: X-14 bathroom cleaner works great. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 -----Original Message----- From: Hofecker, Jennifer L [mailto:jennifer.l.hofecker@Vanderbilt.Edu] Sent: Friday, September 23, 2005 9:24 AM To: histonet Subject: [Histonet] removing silver from counters, etc. Happy Friday everyone. Can somebody remind me what solutions will remove silver nitrate from cabinets, floors, counters, etc.? Bleach just changes these spots to different shades of brown. I remember using solution from hair perms in the past but I am sure that there are other reliable ways. Thanks for any advice. Our lab is starting to resemble a dalmation! You know how silver solution likes to migrate from the coplin jars! Thoughts and prayers go out to Gulf Coast residents. Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From sjchtascp <@t> yahoo.com Fri Sep 23 08:41:14 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 23 08:41:46 2005 Subject: [Histonet] antigen retrevial/ slides Message-ID: <20050923134114.60773.qmail@web90203.mail.scd.yahoo.com> Are there any recommendations for a good adhesive slide to use for IHC/antigen retrevial. Also method for drying these slides Steve --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From DWeil <@t> ciit.org Fri Sep 23 08:44:46 2005 From: DWeil <@t> ciit.org (David Weil) Date: Fri Sep 23 08:45:06 2005 Subject: [Histonet] Ultralow Freezers Message-ID: <12816CB59E68184F9F5F28063E68B0470276B795@xsrvr.ciit.org> I use both types of freezers. I prefer the chest type freezers because you do not have the massive rush of freezing air when you open the door and you avoid having a glacier forming on the inner door. The ice build up on the uprights is especially bad over the summers here, because of high humidity and someone ends up having to defrost the inner door on a regular basis. The only drawback to the chest freezers I have seen is if somebody who is short has to retrieve some samples. Other wise they work great and you do not have to defrost the door. A colleague here has a relatively small chest freezer (approximately 2.5 feet deep x 5 feet long by 3.5 feet tall) made by Revco. The Model # is ULT1090-3-A31. The most common uprights we have now are made by Sanyo (approximately 3.25 feet deep x 3.75 feet wide x 6.75 feet tall) They are a VIP series model # MDF-U72VC. Good luck with your decision. David S Weil BS, HTL (ASCP) CM CIIT Centers For Health Research 6 Davis Dr. RTP, NC 27709-2137 (919) 558-1265 From juan.gutierrez <@t> christushealth.org Fri Sep 23 09:03:06 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 23 09:03:34 2005 Subject: [Histonet] removing silver from counters, etc. Message-ID: I'm not sure about counters, but for skin I've always use potassium ferrocyanide followed by Hypo. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hofecker, Jennifer L Sent: Friday, September 23, 2005 8:24 AM To: histonet Subject: [Histonet] removing silver from counters, etc. Happy Friday everyone. Can somebody remind me what solutions will remove silver nitrate from cabinets, floors, counters, etc.? Bleach just changes these spots to different shades of brown. I remember using solution from hair perms in the past but I am sure that there are other reliable ways. Thanks for any advice. Our lab is starting to resemble a dalmation! You know how silver solution likes to migrate from the coplin jars! Thoughts and prayers go out to Gulf Coast residents. Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Fri Sep 23 09:15:06 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 23 09:20:13 2005 Subject: [Histonet] Antigen retreival on mouse GFAP. In-Reply-To: References: Message-ID: <43340DEA.2080803@umdnj.edu> Hi Susan: I don't know why you would need antigen retrieval for GFAP on mouse brain. Most people I know use DAKO's rabbit anti-cow antibody, catalogue # Z0334. It works very well on both fixed, frozen sections and on formalin-fixed wax embedded tissue. With a Vector ABC Elite kit my dilution for GFAP primary is 1:2000 or even more dilute, depending on how I intensify the DAB. I also get great results with fluorescence. Perhaps your trouble is somewere else? Geoff Susan.Ferrigon@sanofi-aventis.com wrote: >Hi > >Does anyone have any experience with using trypsin as the method of antigen >retreival when staining GFAP on mouse brains??? Any advice on >concentration, temperature and incubation time would be appreciated. > >Thanks > >Susan. > >---------------------------------------------------------- >Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est >exclusivement destin? au(x) destinataire(s), personnes physiques ou >morales, qu'il d?signe. >Il constitue de ce fait une correspondance ? caract?re priv? et peut >contenir des informations confidentielles. >Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser >imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le >d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de >copie. > >This message, including any attachment, is intended for the use of the >individual or entity to which it is addressed. >It is therefore to be considered as a private correspondence which may >contain confidential information. >If you are not the intended recipient, please advise the sender immediately >by reply e.mail and delete this message and any attachment thereto without >retaining a copy. >---------------------------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From carl.hobbs <@t> kcl.ac.uk Fri Sep 23 09:46:29 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 23 09:47:31 2005 Subject: [Histonet] re Hu Ab Message-ID: <007301c5c04d$956cafb0$112b5c9f@Carlos> I use anti Hu C/D on pwax sections of human autopsy brain, after HIER. Molecular Probes A-21276, around 1/3000 dilution factor. Very good results, suprisingly. The Ab reagent also works vwell in pwax sections of chick, mouse and rat. carl From timothy.macatee <@t> med.nyu.edu Fri Sep 23 09:49:28 2005 From: timothy.macatee <@t> med.nyu.edu (Timothy Macatee) Date: Fri Sep 23 09:51:58 2005 Subject: [Histonet] Let's Play solve this problem.... In-Reply-To: Message-ID: Histonetters Assemble! I'm having a recurring problem with my H & E stained slides. The staining is weak, especially away from the edges of the tissue. It looks almost like there is still an opaque paraffin haze to the tissue. It could be a processing problem because it occurs more towards the middle of the section. Is something not getting into the tissue far enough? Is it Fixative (PFA or Formalin), or processing reagents? Or am I just whistling dixie? Tim Macatee NYU Medical Center Research Histology Core From shive003 <@t> umn.edu Fri Sep 23 10:31:07 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 23 10:31:33 2005 Subject: [Histonet] Antigen retreival on mouse GFAP. References: Message-ID: <001901c5c053$d143b0a0$41065486@auxs.umn.edu> In my experience with various mammals, antigen retrieval is not needed for GFAP IHC. I run mine (using Dako's rabbit anti-cow GFAP) at 30 minutes primary incubation (3 ug/ml Ab working concentration), 30 minutes anti-rabbit IgG EnVision+/HRP, and 10 minutes AEC chromogen. Jan Shivers U of MN Vet Diag Lab * ----- Original Message ----- From: To: Sent: Friday, September 23, 2005 5:27 AM Subject: [Histonet] Antigen retreival on mouse GFAP. Hi Does anyone have any experience with using trypsin as the method of antigen retreival when staining GFAP on mouse brains??? Any advice on concentration, temperature and incubation time would be appreciated. Thanks Susan. From shive003 <@t> umn.edu Fri Sep 23 10:34:12 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 23 10:34:58 2005 Subject: [Histonet] antigen retrevial/ slides References: <20050923134114.60773.qmail@web90203.mail.scd.yahoo.com> Message-ID: <002601c5c054$3f932180$41065486@auxs.umn.edu> Poly-L-Lysine. Never had a problem with adherence after HIER when using them. If you coat your own, let them dry overnight (covered) on edge, so excess solution can drain off and not puddle up. Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "Steven Coakley" To: Sent: Friday, September 23, 2005 8:41 AM Subject: [Histonet] antigen retrevial/ slides > Are there any recommendations for a good adhesive slide to use for IHC/antigen retrevial. Also method for drying these slides > > Steve > > > --------------------------------- > Yahoo! for Good > Click here to donate to the Hurricane Katrina relief effort. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gcallis <@t> montana.edu Fri Sep 23 10:41:53 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 23 10:42:20 2005 Subject: [Histonet] Re:sectioning problems In-Reply-To: <20050923052046.8C2919E851@merle.it.northwestern.edu> References: <20050923052046.8C2919E851@merle.it.northwestern.edu> Message-ID: <6.0.0.22.1.20050923092322.01b67910@gemini.msu.montana.edu> First and foremost, if you are having the ribbon pulling up as chuck with block (Point #3) you need to make sure knife angle setting on the microtome is correct. Check your manufacturers instructions on HOW to set a correct knife angle for your particular microtome. it sound as though the clearance angle is incorrect. I don't think the paraffin is the problem, see last comments. You did not say what tissues you are sectioning animal? human? mouse? nor how the tissue is processed? If the tissues are dehydrated, cleared and infiltrated with paraffin - optimally for your particular tissue? then you should experience good sectioning with correct microtome settings. Beware of infiltrating tissues with excessively hot paraffin, 60C is all that we allow with all your animal tisues, over that - I think and with long infiltration times, you dry out and create some sectioning problems. 64C is pretty hot in my estimation for embedding, with ANY paraffin. We set ours at 60C only. You have hot and cold surfaces on your embedding center to help you out, these are usually preset at factory although we can change that with our Sakura embedding center. Too hot is an enemy, as you cook the tissues even more. Even the kind of blades you use will affect how your sectioning goes, and we all have our favorites - be sure to try others on the market!!! Dirrerent brand paraffins will have different sectioning qualities depending on what additives they add to make sectioning easier, etc. Some paraffins are sticky, others are harder, some you have more compression, but with care and correct knife angle setting, one can usually learn to work with ANY paraffin. These additives will cause the paraffins to look different too - opacity, clarity, hardness, ribboning ability, tackiness as you have noticed. Although I like flowers, I favor the brownies!!! At 11:20 PM 9/22/2005, you wrote: >I recently have been having problems pulling ribbons from my microtome and >was >wondering if anyone has advice. > >1.) I was having problems pulling ribbons from newly embeded blocks, but >ones that had >been embeded previously were very easy to pull ribbons from. The new ones >would simply >curl on the blade and wouldn't even grab on to the adjascent sections. > >2.) I changed the paraffin in the embedding station because I noticed all >the temeratures >were set to 64 C when the melting point for the Surgipath paraffin formula >'R' was 56-67C. >Could this have been a problem? > >3.) I switched to an old lot of Fisher embeding medium and now all the >section come off, but >on occasion when the chuck on the mictomome comes back up it seems to (via >static >electricity?) pull the ribbon from the microtome blade and interupt the >ribbon. > >4.) Also, visually the Fisher paraffin blocks look almost transparent >whereas the Surgipath >blocks seem opaque and white. > >I'm relatively new to the histology game and I realize that there is both >art and science >involved, but this is clearly unacceptable behavior from the paraffin, >something is surely >wrong. I'm sure you are all experienced and maybe you have advice? If >you do I would >apreciate it so much. The first and best advice get their choice of FTD >flowers and/ or >homemade brownies. > >Thank you--Tyler Wellington > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Jackie.O'Connor <@t> abbott.com Fri Sep 23 10:50:34 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 23 10:51:09 2005 Subject: [Histonet] Let's Play solve this problem.... Message-ID: Tim - Are there any prizes for playing? For IHC it would automatically set off incomplete fixation alarms. How big are your tissues? What kind of tissue are we talking about? If your tissues are not completely fixed in formalin (or whatever you fix in) they are getting primarily fixed in alcohol. My second instinct would be contamination of your deparaffinization xylene (or clearing agent) in your staining set up, not allowing the hematoxylin to get to the nuclei because there is an invisible coating of paraffin - which of course is then completely removed in the last clearing xylenes. Third - Incomplete processing - smell your block - no really, go ahead - smell your block. Does it smell like any reagent in particular? You shouldn't be able to smell anything if the processing was right. But, I sense myself rambling . . . . Jackie O' Timothy Macatee Sent by: histonet-bounces@lists.utsouthwestern.edu 09/23/2005 09:49 AM To: Histonetters cc: Subject: [Histonet] Let's Play solve this problem.... Histonetters Assemble! I'm having a recurring problem with my H & E stained slides. The staining is weak, especially away from the edges of the tissue. It looks almost like there is still an opaque paraffin haze to the tissue. It could be a processing problem because it occurs more towards the middle of the section. Is something not getting into the tissue far enough? Is it Fixative (PFA or Formalin), or processing reagents? Or am I just whistling dixie? Tim Macatee NYU Medical Center Research Histology Core _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Melissa.Gonzalez <@t> cellgenesys.com Fri Sep 23 10:57:14 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Fri Sep 23 10:58:26 2005 Subject: [Histonet] Re: Antigen retrieval on mouse GFAP Message-ID: Susan, I have to agree with Geoff on Dako's GFAP. It is fantastic and doesn't require Ag retrieval. It picks up astrocytes beautifully in mouse brain. I would definitely recommend DAKO if you aren't using it already. Melissa Melissa A. Gonzalez Histology Cell Genesys 500 Forbes Blvd. South San Francisco, CA 94080 Message: 25 Date: Fri, 23 Sep 2005 10:15:06 -0400 From: Geoff McAuliffe Subject: Re: [Histonet] Antigen retreival on mouse GFAP. To: Susan.Ferrigon@sanofi-aventis.com Cc: histonet@lists.utsouthwestern.edu Message-ID: <43340DEA.2080803@umdnj.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hi Susan: I don't know why you would need antigen retrieval for GFAP on mouse brain. Most people I know use DAKO's rabbit anti-cow antibody, catalogue # Z0334. It works very well on both fixed, frozen sections and on formalin-fixed wax embedded tissue. With a Vector ABC Elite kit my dilution for GFAP primary is 1:2000 or even more dilute, depending on how I intensify the DAB. I also get great results with fluorescence. Perhaps your trouble is somewere else? Geoff Susan.Ferrigon@sanofi-aventis.com wrote: >Hi > >Does anyone have any experience with using trypsin as the method of antigen >retreival when staining GFAP on mouse brains??? Any advice on >concentration, temperature and incubation time would be appreciated. > >Thanks > >Susan. > >---------------------------------------------------------- >Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est >exclusivement destin? au(x) destinataire(s), personnes physiques ou >morales, qu'il d?signe. >Il constitue de ce fait une correspondance ? caract?re priv? et peut >contenir des informations confidentielles. >Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser >imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le >d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de >copie. > >This message, including any attachment, is intended for the use of the >individual or entity to which it is addressed. >It is therefore to be considered as a private correspondence which may >contain confidential information. >If you are not the intended recipient, please advise the sender immediately >by reply e.mail and delete this message and any attachment thereto without >retaining a copy. >---------------------------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 28 **************************************** From gcallis <@t> montana.edu Fri Sep 23 11:02:35 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 23 11:02:59 2005 Subject: [Histonet] removing silver from counters, etc. In-Reply-To: <898D946569A27444B65667A49C0740520754B1@mailbe06.mc.vanderb ilt.edu> References: <898D946569A27444B65667A49C0740520754B1@mailbe06.mc.vanderbilt.edu> Message-ID: <6.0.0.22.1.20050923095708.01b7f9c0@gemini.msu.montana.edu> Put a concentrated solution of i.e 10% or more sodium thiosulfate on top of the silver spot, could be a cloth or paper towel soaked in this, and check the progress. I recovered a damaged blouse this way years ago. I also suggest using tabletop "diapers" or bench protectors (disposable) to prevent spills onto surfaces on the future, a simple paper towel works too. At 07:23 AM 9/23/2005, you wrote: >Happy Friday everyone. >Can somebody remind me what solutions will remove silver nitrate from >cabinets, floors, counters, etc.? >Bleach just changes these spots to different shades of brown. I remember >using solution from hair perms in the past but I am sure that there are >other reliable ways. >Thanks for any advice. Our lab is starting to resemble a dalmation! You >know how silver solution likes to migrate from the coplin jars! > >Thoughts and prayers go out to Gulf Coast residents. > >Jennifer > > > >Jennifer Hofecker, HT (ASCP) >Vanderbilt University Medical Center >Division of Neuropathology >(615) 343-0083 >(615) 343-7089 fax >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Fri Sep 23 11:03:28 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 23 11:03:47 2005 Subject: [Histonet] antigen retrevial/ slides In-Reply-To: <20050923134114.60773.qmail@web90203.mail.scd.yahoo.com> References: <20050923134114.60773.qmail@web90203.mail.scd.yahoo.com> Message-ID: <6.0.0.22.1.20050923100253.01b96650@gemini.msu.montana.edu> What retrieval are you having problems with and what kind of slides do you use now? At 07:41 AM 9/23/2005, you wrote: >Are there any recommendations for a good adhesive slide to use for >IHC/antigen retrevial. Also method for drying these slides > >Steve > > >--------------------------------- >Yahoo! for Good > Click here to donate to the Hurricane Katrina relief effort. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Fri Sep 23 11:04:47 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 23 11:05:15 2005 Subject: "Hypo" is RE: [Histonet] removing silver from counters, etc. In-Reply-To: References: Message-ID: <6.0.0.22.1.20050923100352.01c36d40@gemini.msu.montana.edu> "Hypo" is sodium thiosulfate, folks!! At 08:03 AM 9/23/2005, you wrote: >I'm not sure about counters, but for skin I've always use potassium >ferrocyanide followed by Hypo. > >Juan C. Gutierrez, HT(ASCP) >Histology Laboratory Supervisor >(210)704-2533 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From dellav <@t> musc.edu Fri Sep 23 11:34:28 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 23 11:34:03 2005 Subject: [Histonet] Re:sectioning problems Message-ID: I completely agree with Gayle that your clearance angle sounds like the culprit. However, on the outside chance that you made have changed the brand of blades you use, the facet angle may be different which would also require you to adjust the clearance angle to get the tilt just right to avoid compression or sections lifting from the blade. I'm not convinced that the 64 degrees of your paraffin would have caused the problem you describe. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> Gayle Callis 09/23/05 11:41AM >>> First and foremost, if you are having the ribbon pulling up as chuck with block (Point #3) you need to make sure knife angle setting on the microtome is correct. Check your manufacturers instructions on HOW to set a correct knife angle for your particular microtome. it sound as though the clearance angle is incorrect. I don't think the paraffin is the problem, see last comments. You did not say what tissues you are sectioning animal? human? mouse? nor how the tissue is processed? If the tissues are dehydrated, cleared and infiltrated with paraffin - optimally for your particular tissue? then you should experience good sectioning with correct microtome settings. Beware of infiltrating tissues with excessively hot paraffin, 60C is all that we allow with all your animal tisues, over that - I think and with long infiltration times, you dry out and create some sectioning problems. 64C is pretty hot in my estimation for embedding, with ANY paraffin. We set ours at 60C only. You have hot and cold surfaces on your embedding center to help you out, these are usually preset at factory although we can change that with our Sakura embedding center. Too hot is an enemy, as you cook the tissues even more. Even the kind of blades you use will affect how your sectioning goes, and we all have our favorites - be sure to try others on the market!!! Dirrerent brand paraffins will have different sectioning qualities depending on what additives they add to make sectioning easier, etc. Some paraffins are sticky, others are harder, some you have more compression, but with care and correct knife angle setting, one can usually learn to work with ANY paraffin. These additives will cause the paraffins to look different too - opacity, clarity, hardness, ribboning ability, tackiness as you have noticed. Although I like flowers, I favor the brownies!!! At 11:20 PM 9/22/2005, you wrote: >I recently have been having problems pulling ribbons from my microtome and >was >wondering if anyone has advice. > >1.) I was having problems pulling ribbons from newly embeded blocks, but >ones that had >been embeded previously were very easy to pull ribbons from. The new ones >would simply >curl on the blade and wouldn't even grab on to the adjascent sections. > >2.) I changed the paraffin in the embedding station because I noticed all >the temeratures >were set to 64 C when the melting point for the Surgipath paraffin formula >'R' was 56-67C. >Could this have been a problem? > >3.) I switched to an old lot of Fisher embeding medium and now all the >section come off, but >on occasion when the chuck on the mictomome comes back up it seems to (via >static >electricity?) pull the ribbon from the microtome blade and interupt the >ribbon. > >4.) Also, visually the Fisher paraffin blocks look almost transparent >whereas the Surgipath >blocks seem opaque and white. > >I'm relatively new to the histology game and I realize that there is both >art and science >involved, but this is clearly unacceptable behavior from the paraffin, >something is surely >wrong. I'm sure you are all experienced and maybe you have advice? If >you do I would >apreciate it so much. The first and best advice get their choice of FTD >flowers and/ or >homemade brownies. > >Thank you--Tyler Wellington > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> pathreflab.com Fri Sep 23 12:10:00 2005 From: jnocito <@t> pathreflab.com (Joe Nocito) Date: Fri Sep 23 12:10:34 2005 Subject: [Histonet] Let's Play solve this problem.... In-Reply-To: Message-ID: Tim, Can you list the processing reagents? Are you using formalin or xylene substitutes? Is this a new problem or a chronic one? Have you changed vendors recently for your processing reagents or stains? How often are you changing your processing and staining solutions? Have you changed your processing times? Just wondering Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Timothy Macatee Sent: Friday, September 23, 2005 9:49 AM To: Histonetters Subject: [Histonet] Let's Play solve this problem.... Histonetters Assemble! I'm having a recurring problem with my H & E stained slides. The staining is weak, especially away from the edges of the tissue. It looks almost like there is still an opaque paraffin haze to the tissue. It could be a processing problem because it occurs more towards the middle of the section. Is something not getting into the tissue far enough? Is it Fixative (PFA or Formalin), or processing reagents? Or am I just whistling dixie? Tim Macatee NYU Medical Center Research Histology Core _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mstahl <@t> bvha.org Fri Sep 23 12:35:56 2005 From: mstahl <@t> bvha.org (Stahl, Michael) Date: Fri Sep 23 12:35:05 2005 Subject: [Histonet] http://www.histosearch.com/histonet/Sep05A/RE.HistonetLetsPlaysolvetA.html Message-ID: <4C878E714B21EB4F8EB159777B8822EE5B6767@bvfyms01.net.bvha.org> Ok ill play along,, sounds to me like your clearing agent needs changed. From Heather.A.Harper <@t> pcola.med.navy.mil Fri Sep 23 12:44:50 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Fri Sep 23 12:57:11 2005 Subject: [Histonet] Histonetters Assemble Message-ID: <807FE48C5A7CC940B973B58D32E701431B13B446@nhpens-exch1.pcola.med.navy.mil> To me it sounds like you are having paraffin problems and not fixation. Maybe you are not leaving your slides in the oven long enough, or the paraffin from the embedding station has water or something else in it creating this opaque film. -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 22, Issue 29 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. re Hu Ab (Carl Hobbs) 2. Let's Play solve this problem.... (Timothy Macatee) 3. Re: Antigen retreival on mouse GFAP. (Jan Shivers) 4. Re: antigen retrevial/ slides (Jan Shivers) 5. Re:sectioning problems (Gayle Callis) 6. Re: Let's Play solve this problem.... (Jackie M O'Connor) 7. Re: Antigen retrieval on mouse GFAP (Melissa Gonzalez) 8. Re: removing silver from counters, etc. (Gayle Callis) 9. Re: antigen retrevial/ slides (Gayle Callis) 10. "Hypo" is RE: [Histonet] removing silver from counters, etc. (Gayle Callis) 11. Re:sectioning problems (Vinnie Della Speranza) ---------------------------------------------------------------------- Message: 1 Date: Fri, 23 Sep 2005 15:46:29 +0100 From: "Carl Hobbs" Subject: [Histonet] re Hu Ab To: Message-ID: <007301c5c04d$956cafb0$112b5c9f@Carlos> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original I use anti Hu C/D on pwax sections of human autopsy brain, after HIER. Molecular Probes A-21276, around 1/3000 dilution factor. Very good results, suprisingly. The Ab reagent also works vwell in pwax sections of chick, mouse and rat. carl ------------------------------ Message: 2 Date: Fri, 23 Sep 2005 10:49:28 -0400 From: Timothy Macatee Subject: [Histonet] Let's Play solve this problem.... To: Histonetters Message-ID: Content-Type: text/plain; charset=US-ASCII Histonetters Assemble! I'm having a recurring problem with my H & E stained slides. The staining is weak, especially away from the edges of the tissue. It looks almost like there is still an opaque paraffin haze to the tissue. It could be a processing problem because it occurs more towards the middle of the section. Is something not getting into the tissue far enough? Is it Fixative (PFA or Formalin), or processing reagents? Or am I just whistling dixie? Tim Macatee NYU Medical Center Research Histology Core ------------------------------ Message: 3 Date: Fri, 23 Sep 2005 10:31:07 -0500 From: "Jan Shivers" Subject: Re: [Histonet] Antigen retreival on mouse GFAP. To: Cc: histonet Message-ID: <001901c5c053$d143b0a0$41065486@auxs.umn.edu> Content-Type: text/plain; charset="iso-8859-1" In my experience with various mammals, antigen retrieval is not needed for GFAP IHC. I run mine (using Dako's rabbit anti-cow GFAP) at 30 minutes primary incubation (3 ug/ml Ab working concentration), 30 minutes anti-rabbit IgG EnVision+/HRP, and 10 minutes AEC chromogen. Jan Shivers U of MN Vet Diag Lab * ----- Original Message ----- From: To: Sent: Friday, September 23, 2005 5:27 AM Subject: [Histonet] Antigen retreival on mouse GFAP. Hi Does anyone have any experience with using trypsin as the method of antigen retreival when staining GFAP on mouse brains??? Any advice on concentration, temperature and incubation time would be appreciated. Thanks Susan. ------------------------------ Message: 4 Date: Fri, 23 Sep 2005 10:34:12 -0500 From: "Jan Shivers" Subject: Re: [Histonet] antigen retrevial/ slides To: "Steven Coakley" Cc: histonet Message-ID: <002601c5c054$3f932180$41065486@auxs.umn.edu> Content-Type: text/plain; charset="iso-8859-1" Poly-L-Lysine. Never had a problem with adherence after HIER when using them. If you coat your own, let them dry overnight (covered) on edge, so excess solution can drain off and not puddle up. Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "Steven Coakley" To: Sent: Friday, September 23, 2005 8:41 AM Subject: [Histonet] antigen retrevial/ slides > Are there any recommendations for a good adhesive slide to use for IHC/antigen retrevial. Also method for drying these slides > > Steve > > > --------------------------------- > Yahoo! for Good > Click here to donate to the Hurricane Katrina relief effort. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 5 Date: Fri, 23 Sep 2005 09:41:53 -0600 From: Gayle Callis Subject: [Histonet] Re:sectioning problems To: "Tyler W." , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050923092322.01b67910@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed First and foremost, if you are having the ribbon pulling up as chuck with block (Point #3) you need to make sure knife angle setting on the microtome is correct. Check your manufacturers instructions on HOW to set a correct knife angle for your particular microtome. it sound as though the clearance angle is incorrect. I don't think the paraffin is the problem, see last comments. You did not say what tissues you are sectioning animal? human? mouse? nor how the tissue is processed? If the tissues are dehydrated, cleared and infiltrated with paraffin - optimally for your particular tissue? then you should experience good sectioning with correct microtome settings. Beware of infiltrating tissues with excessively hot paraffin, 60C is all that we allow with all your animal tisues, over that - I think and with long infiltration times, you dry out and create some sectioning problems. 64C is pretty hot in my estimation for embedding, with ANY paraffin. We set ours at 60C only. You have hot and cold surfaces on your embedding center to help you out, these are usually preset at factory although we can change that with our Sakura embedding center. Too hot is an enemy, as you cook the tissues even more. Even the kind of blades you use will affect how your sectioning goes, and we all have our favorites - be sure to try others on the market!!! Dirrerent brand paraffins will have different sectioning qualities depending on what additives they add to make sectioning easier, etc. Some paraffins are sticky, others are harder, some you have more compression, but with care and correct knife angle setting, one can usually learn to work with ANY paraffin. These additives will cause the paraffins to look different too - opacity, clarity, hardness, ribboning ability, tackiness as you have noticed. Although I like flowers, I favor the brownies!!! At 11:20 PM 9/22/2005, you wrote: >I recently have been having problems pulling ribbons from my microtome and >was >wondering if anyone has advice. > >1.) I was having problems pulling ribbons from newly embeded blocks, but >ones that had >been embeded previously were very easy to pull ribbons from. The new ones >would simply >curl on the blade and wouldn't even grab on to the adjascent sections. > >2.) I changed the paraffin in the embedding station because I noticed all >the temeratures >were set to 64 C when the melting point for the Surgipath paraffin formula >'R' was 56-67C. >Could this have been a problem? > >3.) I switched to an old lot of Fisher embeding medium and now all the >section come off, but >on occasion when the chuck on the mictomome comes back up it seems to (via >static >electricity?) pull the ribbon from the microtome blade and interupt the >ribbon. > >4.) Also, visually the Fisher paraffin blocks look almost transparent >whereas the Surgipath >blocks seem opaque and white. > >I'm relatively new to the histology game and I realize that there is both >art and science >involved, but this is clearly unacceptable behavior from the paraffin, >something is surely >wrong. I'm sure you are all experienced and maybe you have advice? If >you do I would >apreciate it so much. The first and best advice get their choice of FTD >flowers and/ or >homemade brownies. > >Thank you--Tyler Wellington > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 6 Date: Fri, 23 Sep 2005 10:50:34 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Let's Play solve this problem.... To: Timothy Macatee Cc: Histonetters , histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Tim - Are there any prizes for playing? For IHC it would automatically set off incomplete fixation alarms. How big are your tissues? What kind of tissue are we talking about? If your tissues are not completely fixed in formalin (or whatever you fix in) they are getting primarily fixed in alcohol. My second instinct would be contamination of your deparaffinization xylene (or clearing agent) in your staining set up, not allowing the hematoxylin to get to the nuclei because there is an invisible coating of paraffin - which of course is then completely removed in the last clearing xylenes. Third - Incomplete processing - smell your block - no really, go ahead - smell your block. Does it smell like any reagent in particular? You shouldn't be able to smell anything if the processing was right. But, I sense myself rambling . . . . Jackie O' Timothy Macatee Sent by: histonet-bounces@lists.utsouthwestern.edu 09/23/2005 09:49 AM To: Histonetters cc: Subject: [Histonet] Let's Play solve this problem.... Histonetters Assemble! I'm having a recurring problem with my H & E stained slides. The staining is weak, especially away from the edges of the tissue. It looks almost like there is still an opaque paraffin haze to the tissue. It could be a processing problem because it occurs more towards the middle of the section. Is something not getting into the tissue far enough? Is it Fixative (PFA or Formalin), or processing reagents? Or am I just whistling dixie? Tim Macatee NYU Medical Center Research Histology Core _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Fri, 23 Sep 2005 08:57:14 -0700 From: "Melissa Gonzalez" Subject: [Histonet] Re: Antigen retrieval on mouse GFAP To: Cc: Susan.Ferrigon@sanofi-aventis.com Message-ID: Content-Type: text/plain; charset="iso-8859-1" Susan, I have to agree with Geoff on Dako's GFAP. It is fantastic and doesn't require Ag retrieval. It picks up astrocytes beautifully in mouse brain. I would definitely recommend DAKO if you aren't using it already. Melissa Melissa A. Gonzalez Histology Cell Genesys 500 Forbes Blvd. South San Francisco, CA 94080 Message: 25 Date: Fri, 23 Sep 2005 10:15:06 -0400 From: Geoff McAuliffe Subject: Re: [Histonet] Antigen retreival on mouse GFAP. To: Susan.Ferrigon@sanofi-aventis.com Cc: histonet@lists.utsouthwestern.edu Message-ID: <43340DEA.2080803@umdnj.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hi Susan: I don't know why you would need antigen retrieval for GFAP on mouse brain. Most people I know use DAKO's rabbit anti-cow antibody, catalogue # Z0334. It works very well on both fixed, frozen sections and on formalin-fixed wax embedded tissue. With a Vector ABC Elite kit my dilution for GFAP primary is 1:2000 or even more dilute, depending on how I intensify the DAB. I also get great results with fluorescence. Perhaps your trouble is somewere else? Geoff Susan.Ferrigon@sanofi-aventis.com wrote: >Hi > >Does anyone have any experience with using trypsin as the method of antigen >retreival when staining GFAP on mouse brains??? Any advice on >concentration, temperature and incubation time would be appreciated. > >Thanks > >Susan. > >---------------------------------------------------------- >Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est >exclusivement destin? au(x) destinataire(s), personnes physiques ou >morales, qu'il d?signe. >Il constitue de ce fait une correspondance ? caract?re priv? et peut >contenir des informations confidentielles. >Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser >imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le >d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de >copie. > >This message, including any attachment, is intended for the use of the >individual or entity to which it is addressed. >It is therefore to be considered as a private correspondence which may >contain confidential information. >If you are not the intended recipient, please advise the sender immediately >by reply e.mail and delete this message and any attachment thereto without >retaining a copy. >---------------------------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 28 **************************************** ------------------------------ Message: 8 Date: Fri, 23 Sep 2005 10:02:35 -0600 From: Gayle Callis Subject: Re: [Histonet] removing silver from counters, etc. To: "Hofecker, Jennifer L" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050923095708.01b7f9c0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Put a concentrated solution of i.e 10% or more sodium thiosulfate on top of the silver spot, could be a cloth or paper towel soaked in this, and check the progress. I recovered a damaged blouse this way years ago. I also suggest using tabletop "diapers" or bench protectors (disposable) to prevent spills onto surfaces on the future, a simple paper towel works too. At 07:23 AM 9/23/2005, you wrote: >Happy Friday everyone. >Can somebody remind me what solutions will remove silver nitrate from >cabinets, floors, counters, etc.? >Bleach just changes these spots to different shades of brown. I remember >using solution from hair perms in the past but I am sure that there are >other reliable ways. >Thanks for any advice. Our lab is starting to resemble a dalmation! You >know how silver solution likes to migrate from the coplin jars! > >Thoughts and prayers go out to Gulf Coast residents. > >Jennifer > > > >Jennifer Hofecker, HT (ASCP) >Vanderbilt University Medical Center >Division of Neuropathology >(615) 343-0083 >(615) 343-7089 fax >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 9 Date: Fri, 23 Sep 2005 10:03:28 -0600 From: Gayle Callis Subject: Re: [Histonet] antigen retrevial/ slides To: Steven Coakley , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050923100253.01b96650@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed What retrieval are you having problems with and what kind of slides do you use now? At 07:41 AM 9/23/2005, you wrote: >Are there any recommendations for a good adhesive slide to use for >IHC/antigen retrevial. Also method for drying these slides > >Steve > > >--------------------------------- >Yahoo! for Good > Click here to donate to the Hurricane Katrina relief effort. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 10 Date: Fri, 23 Sep 2005 10:04:47 -0600 From: Gayle Callis Subject: "Hypo" is RE: [Histonet] removing silver from counters, etc. To: "GUTIERREZ, JUAN" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050923100352.01c36d40@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed "Hypo" is sodium thiosulfate, folks!! At 08:03 AM 9/23/2005, you wrote: >I'm not sure about counters, but for skin I've always use potassium >ferrocyanide followed by Hypo. > >Juan C. Gutierrez, HT(ASCP) >Histology Laboratory Supervisor >(210)704-2533 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 11 Date: Fri, 23 Sep 2005 12:34:28 -0400 From: "Vinnie Della Speranza" Subject: [Histonet] Re:sectioning problems To: , , Message-ID: Content-Type: text/plain; charset=US-ASCII I completely agree with Gayle that your clearance angle sounds like the culprit. However, on the outside chance that you made have changed the brand of blades you use, the facet angle may be different which would also require you to adjust the clearance angle to get the tilt just right to avoid compression or sections lifting from the blade. I'm not convinced that the 64 degrees of your paraffin would have caused the problem you describe. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> Gayle Callis 09/23/05 11:41AM >>> First and foremost, if you are having the ribbon pulling up as chuck with block (Point #3) you need to make sure knife angle setting on the microtome is correct. Check your manufacturers instructions on HOW to set a correct knife angle for your particular microtome. it sound as though the clearance angle is incorrect. I don't think the paraffin is the problem, see last comments. You did not say what tissues you are sectioning animal? human? mouse? nor how the tissue is processed? If the tissues are dehydrated, cleared and infiltrated with paraffin - optimally for your particular tissue? then you should experience good sectioning with correct microtome settings. Beware of infiltrating tissues with excessively hot paraffin, 60C is all that we allow with all your animal tisues, over that - I think and with long infiltration times, you dry out and create some sectioning problems. 64C is pretty hot in my estimation for embedding, with ANY paraffin. We set ours at 60C only. You have hot and cold surfaces on your embedding center to help you out, these are usually preset at factory although we can change that with our Sakura embedding center. Too hot is an enemy, as you cook the tissues even more. Even the kind of blades you use will affect how your sectioning goes, and we all have our favorites - be sure to try others on the market!!! Dirrerent brand paraffins will have different sectioning qualities depending on what additives they add to make sectioning easier, etc. Some paraffins are sticky, others are harder, some you have more compression, but with care and correct knife angle setting, one can usually learn to work with ANY paraffin. These additives will cause the paraffins to look different too - opacity, clarity, hardness, ribboning ability, tackiness as you have noticed. Although I like flowers, I favor the brownies!!! At 11:20 PM 9/22/2005, you wrote: >I recently have been having problems pulling ribbons from my microtome and >was >wondering if anyone has advice. > >1.) I was having problems pulling ribbons from newly embeded blocks, but >ones that had >been embeded previously were very easy to pull ribbons from. The new ones >would simply >curl on the blade and wouldn't even grab on to the adjascent sections. > >2.) I changed the paraffin in the embedding station because I noticed all >the temeratures >were set to 64 C when the melting point for the Surgipath paraffin formula >'R' was 56-67C. >Could this have been a problem? > >3.) I switched to an old lot of Fisher embeding medium and now all the >section come off, but >on occasion when the chuck on the mictomome comes back up it seems to (via >static >electricity?) pull the ribbon from the microtome blade and interupt the >ribbon. > >4.) Also, visually the Fisher paraffin blocks look almost transparent >whereas the Surgipath >blocks seem opaque and white. > >I'm relatively new to the histology game and I realize that there is both >art and science >involved, but this is clearly unacceptable behavior from the paraffin, >something is surely >wrong. I'm sure you are all experienced and maybe you have advice? If >you do I would >apreciate it so much. The first and best advice get their choice of FTD >flowers and/ or >homemade brownies. > >Thank you--Tyler Wellington > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 29 **************************************** From cwscouten <@t> myneurolab.com Fri Sep 23 14:01:07 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 23 14:02:09 2005 Subject: [Histonet] cryostat decontamination Message-ID: <5784D843593D874C93E9BADCB87342AB916655@tpiserver03.Coretech-holdings.com> Consider a self decontaminating cryostat, or use the decontaminant used in it. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=4 75102&catdesc=Histology+Equipment&CatThreeID=732&CatOneID=4&subcatdesc=C ryostats&idsubcategory=182 Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Wednesday, September 21, 2005 9:09 AM To: Marirose.Satterfield@MercyMemorial.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] cryostat decontamination I shut down the cryo and decontaminate with Santimaster IV which is a broad spectrum and non corrosive, I leave it on for about 10 minutes, then I use a 70% ethanol, and then I use an absolute. Robyn OHSU >>> "Satterfield, Marirose" >>> 9/21/2005 6:54 AM >>> CAP Checklist Question ANP.24250 Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontaminated records evident? They note that the interior of the cryostat can be decontaminated with 70% ethanol. Later on they say it should be defrosted and decontaminated with a tuberculocidal disinfectant at an interval appropriate for the institution. Are they suggesting that after each use it get decontaminated with the 70% ethanol and only periodically shut down unit and use the TB disinfectant? Thanks M Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Fri Sep 23 14:17:24 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 23 14:18:10 2005 Subject: [Histonet] Frozen sections on cryoprotected CNS Message-ID: <5784D843593D874C93E9BADCB87342AB916657@tpiserver03.Coretech-holdings.com> You didn't mention, how long does the brain sit in the 30% sucrose? This should be 3-7 days, or until it sinks, indicating full penetration. Partial penetration might cause the symptoms you see. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Thomas Crowell Sent: Tuesday, September 20, 2005 3:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozen sections on cryoprotected CNS Dear neurohistology experts, Our laboratory is experiencing great frustration in preparing 10 to 20 micron cryosections from rat and mouse brains that have been cryoprotected in 30% sucrose. The exact sequence of specimen preparation is perfusion fix with 4%PFA, drop fix in PFA for an additional 8-10 hours, transfer to 30% sucrose in PBS, before embedding in OCT. Two events occur that make sectioning difficult. 1:( the OCT does not bond well to the sample causing separation. 2 :( sections that are clearly laying flat on the cryoplate blade holder become distorted and wrinkled when thaw mounted onto the slide (Surgipath X-tra), and do not spread out well on the slide as it dries leaving lots of folds and bubbles in the tissue section. We have tried leaving the cryoprotected sample in OCT at 4C overnight to no avail, and are considering trying some gradients of sucrose, say 5%, 10%, 15% and 20%. As the neurohistolgy component to this laboratory has dramatically increased, so has our frustration level; any help or suggestions to this problem would be greatly appreciated! Thanks Tom Crowell BiogenIdec Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From plaurie <@t> benaroyaresearch.org Fri Sep 23 14:28:35 2005 From: plaurie <@t> benaroyaresearch.org (Patrick Laurie) Date: Fri Sep 23 14:29:06 2005 Subject: [Histonet] RNAse decontamination in cryostats Message-ID: My question is similar to the current topic of discussion about decontaminating cryostats, except that I need to remove any potential RNAse activity. I have been cleaning the disposable blade and the blade holder with 70% alcohol, which hasn't been shown to eliminate the pernicious little buggers, but the commercial products such as RNAse away (Ambion) etc, are water based and would freeze on the blade/blade holder which would make me suspicious of their effectiveness. I am not able to continuously thaw the cryostat and then use the products, as I would like to do daily. I am wondering if anyone has any ideas or potential products that would work. Thanks Patrick Laurie, HT (ASCP) Neurogenomics Laboratory Benaroya Research Institute 1201 9th Ave Seattle, Wa 98101 (206) 341-0681 From TJJ <@t> Stowers-Institute.org Fri Sep 23 14:43:31 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 23 14:44:03 2005 Subject: [Histonet] Re: Integrin beta1 / CD29 -- cell surface or not?? Message-ID: Gustave, I can't offer a direct answer with regards to the molecular characteristics of CD29, but instead can just give specifics to IHC techniques. I would expect membrane staining using this particular antibody. If you are getting nuclear staining, I would recommend taking a look at your retrieval technique. It would be you are "over-retrieving" for this antibody in your samples. Many antibodies that errantly stain nuclei end up needing less antigen unmasking, and/or require further dilution. Make sure your secondary antibody is adsorbed to mouse. Rat and mouse are similar and sometimes anti-rat secondaries can cross react in mouse. (I doubt this is the cause of your problems). Perhaps you can work this up using control tissues instead of your sample; this marker is known to stain lymphocytes, so perhaps thymus or other lymphoid material might suffice and see if you get the same staining patterns. Finally, you might consider using a different fixative, or try staining on frozen section and see if you get the same results. Good luck! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From vazquezr <@t> ohsu.edu Fri Sep 23 15:52:30 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 23 15:53:17 2005 Subject: [Histonet] cryostat decontamination Message-ID: That is a good idea. I won't be getting one for about a year from now. We are going to be moving into a new facility and will be purchasing some new stuff. And a self -decon is on my list. Robyn OHSU >>> "Charles Scouten" 9/23/2005 12:01 PM >>> Consider a self decontaminating cryostat, or use the decontaminant used in it. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=4 75102&catdesc=Histology+Equipment&CatThreeID=732&CatOneID=4&subcatdesc=C ryostats&idsubcategory=182 Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Wednesday, September 21, 2005 9:09 AM To: Marirose.Satterfield@MercyMemorial.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] cryostat decontamination I shut down the cryo and decontaminate with Santimaster IV which is a broad spectrum and non corrosive, I leave it on for about 10 minutes, then I use a 70% ethanol, and then I use an absolute. Robyn OHSU >>> "Satterfield, Marirose" >>> 9/21/2005 6:54 AM >>> CAP Checklist Question ANP.24250 Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontaminated records evident? They note that the interior of the cryostat can be decontaminated with 70% ethanol. Later on they say it should be defrosted and decontaminated with a tuberculocidal disinfectant at an interval appropriate for the institution. Are they suggesting that after each use it get decontaminated with the 70% ethanol and only periodically shut down unit and use the TB disinfectant? Thanks M Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dellav <@t> musc.edu Fri Sep 23 16:39:29 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 23 16:38:54 2005 Subject: [Histonet] cryostat decontamination Message-ID: the implication here is that interior surfaces of the cryostat may be wiped down with 70% ethanol while the unit is cold, in between cases/uses or once daily, however you deem appropriate. full contamination with a tuberculocidal is to be done on a schedule you deem appropriate, which would require taking the unit out of service and returned to room temperature for the disinfecting procedure. others on the list will be more familiar than I with the disinfecting units offered by some manufacturers. I believe one uses UV light although I'd wanted to see some reports in the literature verifying the efficacy of UV at cryostat temperatures which I would assume the manufacturer could provide .. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Satterfield, Marirose" 09/21/05 09:54AM >>> CAP Checklist Question ANP.24250 Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontaminated records evident? They note that the interior of the cryostat can be decontaminated with 70% ethanol. Later on they say it should be defrosted and decontaminated with a tuberculocidal disinfectant at an interval appropriate for the institution. Are they suggesting that after each use it get decontaminated with the 70% ethanol and only periodically shut down unit and use the TB disinfectant? Thanks M Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dellav <@t> musc.edu Fri Sep 23 16:41:34 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 23 16:41:01 2005 Subject: [Histonet] cryostat decontamination Message-ID: can someone provide info on which manufacturers offer self- decontaminating units? Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Robyn Vazquez" 09/23/05 04:52PM >>> That is a good idea. I won't be getting one for about a year from now. We are going to be moving into a new facility and will be purchasing some new stuff. And a self -decon is on my list. Robyn OHSU >>> "Charles Scouten" 9/23/2005 12:01 PM >>> Consider a self decontaminating cryostat, or use the decontaminant used in it. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=4 75102&catdesc=Histology+Equipment&CatThreeID=732&CatOneID=4&subcatdesc=C ryostats&idsubcategory=182 Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Wednesday, September 21, 2005 9:09 AM To: Marirose.Satterfield@MercyMemorial.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] cryostat decontamination I shut down the cryo and decontaminate with Santimaster IV which is a broad spectrum and non corrosive, I leave it on for about 10 minutes, then I use a 70% ethanol, and then I use an absolute. Robyn OHSU >>> "Satterfield, Marirose" >>> 9/21/2005 6:54 AM >>> CAP Checklist Question ANP.24250 Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontaminated records evident? They note that the interior of the cryostat can be decontaminated with 70% ethanol. Later on they say it should be defrosted and decontaminated with a tuberculocidal disinfectant at an interval appropriate for the institution. Are they suggesting that after each use it get decontaminated with the 70% ethanol and only periodically shut down unit and use the TB disinfectant? Thanks M Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Fri Sep 23 16:47:26 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 23 16:48:22 2005 Subject: [Histonet] cryostat decontamination Message-ID: <5784D843593D874C93E9BADCB87342AB91665E@tpiserver03.Coretech-holdings.com> The Vibratome Decontaminating cryostat turns into a dishwasher, and jet sprays a solution of Perascope (periacetic acid plus other things) around the fast thawed interior. No UV. Then a water solution that rinses it all out and down the drain, then cold again. All automatically. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vinnie Della Speranza Sent: Friday, September 23, 2005 4:39 PM To: Marirose.Satterfield@MercyMemorial.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] cryostat decontamination the implication here is that interior surfaces of the cryostat may be wiped down with 70% ethanol while the unit is cold, in between cases/uses or once daily, however you deem appropriate. full contamination with a tuberculocidal is to be done on a schedule you deem appropriate, which would require taking the unit out of service and returned to room temperature for the disinfecting procedure. others on the list will be more familiar than I with the disinfecting units offered by some manufacturers. I believe one uses UV light although I'd wanted to see some reports in the literature verifying the efficacy of UV at cryostat temperatures which I would assume the manufacturer could provide .. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Satterfield, Marirose" >>> 09/21/05 09:54AM >>> CAP Checklist Question ANP.24250 Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontaminated records evident? They note that the interior of the cryostat can be decontaminated with 70% ethanol. Later on they say it should be defrosted and decontaminated with a tuberculocidal disinfectant at an interval appropriate for the institution. Are they suggesting that after each use it get decontaminated with the 70% ethanol and only periodically shut down unit and use the TB disinfectant? Thanks M Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From maria <@t> ski.org Fri Sep 23 17:19:40 2005 From: maria <@t> ski.org (Maria Mejia) Date: Fri Sep 23 17:20:09 2005 Subject: [Histonet] antigen retrieval for IHC Message-ID: <43347F7C.3070109@ski.org> Hello, I believe it was sometime in June of this year that there was a discussion on the histonet regarding "the simplest form" of reversing the effects for formalin fixation on tissue was to wash the tissue well (before) processing. Well, I just read the article "Antigen retrieval IHC: Past, Present, & Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google this article). It's a very interesting! Anyway, the article has a section titled non-heating AR method stating (this) same simple method by (Puchtler & Meloan). It also goes on to say that Elias JM (1990) adopted this method routinely by storing deparaffinized in fresh changes of 10% sucrose/PBS @ 4C overnight (before) IHC. My questions are, does anyone know or tried this method used by Elias? And, can anyone explain the mechanism of 10% sucrose/PBS play in this AR method? Just curious! Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 From hmorrow <@t> cogeco.ca Fri Sep 23 20:25:59 2005 From: hmorrow <@t> cogeco.ca (Marg & Vern Morrow) Date: Fri Sep 23 20:26:23 2005 Subject: [Histonet] C4d Message-ID: <000601c5c0a6$ebc3a9a0$305fe218@your4o5qm9xfjg> Our IP Pathologist's have recently decided to add C4d to to our renal protocol. Is anyone out there using this antibody? Do you use the direct or the in-direct method? What antibody do you use? Tx Marg Morrow From e.kaplan <@t> uaeu.ac.ae Fri Sep 23 22:28:25 2005 From: e.kaplan <@t> uaeu.ac.ae (Evelyn Kaplan) Date: Fri Sep 23 22:28:23 2005 Subject: [Histonet] Plants in lab In-Reply-To: <5.1.0.14.0.20050923043138.00c3da90@mail.jcu.edu.au> Message-ID: <000001c5c0b8$06051900$7c14a00a@aa.uaeu.ac.ae> Rather interestingly, some of the best African violets I have ever seen were grown in the cytology section of the laboratory in Sultan Qaboos University Hospital. Beautiful blooms all year round................. Regards, Evelyn Kaplan B.Sc. MLT Coordinator, Department of Medical Education, Faculty of Medicine & Health Sciences, PO Box 17666, United Arab Emirates University, Al Ain, UAE. Tel: 00 9713 7137216 (O) 00 971 0504717457 (Mobile) 00 9713 7672167 (Fax) e.kaplan@uaeu.ac.ae -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie Reilly Sent: Thursday, September 22, 2005 10:34 PM To: Marshall Terry Dr, Consultant Histopathologist; Cindy DuBois; tmmrosla@healtheast.org Cc: Histonet Subject: RE: [Histonet] Plants in lab Why not a Logwood tree, Haematoxylum campechianum, they make great bonsai specimens? Regards, Laurie. At 05:57 PM 22/09/2005 +0100, Marshall Terry Dr, Consultant Histopathologist wrote: >I'm all for plants in he lab. (or anywhere). > >A Norfolk pine seems a tad extravagant. I've seen them about 20 metres >tall! > >Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path > Consultant Pathologist > Rotherham General Hospital > South Yorkshire > England > terry.marshall@rothgen.nhs.uk > >-----Original Message----- >From: Cindy DuBois [mailto:dpahisto@yahoo.com] >Sent: 22 September 2005 17:51 >To: tmmrosla@healtheast.org >Cc: Histonet >Subject: [Histonet] Plants in lab > > >We have several plants in our lab. African Violets seem to do >extremely >well and bloom all year round. It adds some nice color to our environment. > >Cindy DuBois >Delta Pathology Assoc. >Stockton CA > > >--------------------------------- >Yahoo! for Good > Click here to donate to the Hurricane Katrina relief effort. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Sat Sep 24 10:56:01 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Sat Sep 24 10:56:22 2005 Subject: [Histonet] RNAse decontamination in cryostats Message-ID: Laurie, I have not done this for a number of years but what I did was to use the RNase-ZAP and then follow with a fresh - newly opened bottle of 100% ETOH - remember the most common source of RNase is usually your hands so wear gloves. Seemed to work well. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Patrick Laurie [mailto:plaurie@benaroyaresearch.org] Sent: Friday, September 23, 2005 12:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RNAse decontamination in cryostats My question is similar to the current topic of discussion about decontaminating cryostats, except that I need to remove any potential RNAse activity. I have been cleaning the disposable blade and the blade holder with 70% alcohol, which hasn't been shown to eliminate the pernicious little buggers, but the commercial products such as RNAse away (Ambion) etc, are water based and would freeze on the blade/blade holder which would make me suspicious of their effectiveness. I am not able to continuously thaw the cryostat and then use the products, as I would like to do daily. I am wondering if anyone has any ideas or potential products that would work. Thanks Patrick Laurie, HT (ASCP) Neurogenomics Laboratory Benaroya Research Institute 1201 9th Ave Seattle, Wa 98101 (206) 341-0681 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histomjans <@t> yahoo.com Sat Sep 24 13:08:36 2005 From: histomjans <@t> yahoo.com (Melissa Jans) Date: Sat Sep 24 13:08:56 2005 Subject: [Histonet] lab design Message-ID: <20050924180837.32315.qmail@web54709.mail.yahoo.com> I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From jstaruk <@t> masshistology.com Sat Sep 24 19:59:22 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Sat Sep 24 20:01:46 2005 Subject: [Histonet] lab design In-Reply-To: <20050924180837.32315.qmail@web54709.mail.yahoo.com> Message-ID: <0INC001DZLIAFRF3@vms046.mailsrvcs.net> This is exactly what I did when I designed our new facilities. I built a flame-resistant room which is well-ventilated directly to the outside. The ventilation unit is adjustable for varying amounts of cubic feet per minute air exchange. This unit is always on. There is a special cabinet for flammable chemicals, acid-resistant bench tops for the tissue processors and multiple, adjustable shelves for slide drying in this room. The solid wood passage door is always closed. I also built a fume hood directly on the other side of one of these walls and placed a blower unit (from an old, charcoal bench-top fume hood) through the wall. This is where we coverslip. When the blower is on, it sucks away all of the xylene-laden air into the vented room which, in turn, is vented directly to the outside. There is never any xylene fumes in the lab. On our main homepage, there's a link to photos of our new lab. Here, you will see the ventilated room as well as the coverslipping fume hood on the other side of the ventilated room. Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Melissa Jans Sent: Saturday, September 24, 2005 2:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab design I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vanann702 <@t> skmc.gov.ae Sun Sep 25 05:57:20 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Sun Sep 25 05:57:01 2005 Subject: [Histonet] unsubscribe Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CC08@SKMCEMAIL.skmc.gov.ae> From vanann702 <@t> skmc.gov.ae Sun Sep 25 05:57:20 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Sun Sep 25 05:57:04 2005 Subject: [Histonet] unsubscribe Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CC08@SKMCEMAIL.skmc.gov.ae> From jqb7 <@t> cdc.gov Sun Sep 25 07:22:08 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Sun Sep 25 07:22:27 2005 Subject: [Histonet] lab design Message-ID: Hi Jan: Currently we have a separate room containing our tissue processors and embedding centers. We also have a large flammable storage cabinet in this room. We have a separate room with our alcohol/solvent recycler and there is a large flammable storage cabinet in there as well. We have 2 smaller flammable cabinets in routine lab space as well. We are moving into a brand-new lab building this fall. In the new lab we also will have separate rooms; one for the processors and embedding cetners and another for recycling. I do not think there is flammable storage in the new processing room (it's pretty small) but there is in the recycling room. I believe there will be at least one under-counter flammable storage in a main lab space as well. Jeanine Bartlett CDC, Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Melissa Jans Sent: Sat 9/24/2005 2:08 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] lab design I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tlclab <@t> free.fr Sun Sep 25 10:03:01 2005 From: tlclab <@t> free.fr (TLCLab) Date: Sun Sep 25 10:03:31 2005 Subject: [Histonet] Mellitin Message-ID: <4336BC25.9050801@free.fr> Hello, Is there any - strict - histochemical method to caracterise mellitin (meaning not an immunohistochemical method) ? Regards, Florent. From AnthonyH <@t> chw.edu.au Sun Sep 25 18:33:09 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Sep 25 18:33:55 2005 Subject: [Histonet] plants in histology lab Message-ID: Sue, The reference is from The Sydney Morning Herald February 6th, 1997, an old edition but was an abstract from the book: "Eco-Friendly Houseplants" by B.C. Wolverton, published by Weidenfeld & Nicholson. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: O'Brien, Sue [mailto:sobrien@bthosp.com] Sent: Friday, 23 September 2005 1:31 AM To: Tony Henwood Subject: RE: [Histonet] plants in histology lab Tony, would you by any chance have references for that? I was aware of spider plants being useful for removing formalin fumes, but this is the first I've heard about the others you mention. (Are you by any chance a horticulturist as well?) Thank-you, Sue O'Brien, BS, HT/HTL (ASCP) Burdette Tomlin Memorial Hospital Cape may Court House, NJ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tony Henwood Sent: Wednesday, September 21, 2005 7:57 PM To: 'Mrosla, Tina M'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] plants in histology lab The plants I would suggest are: Peace Lily (Spathiphyllum) Gerbera (Gerbera jamesonii) Chrysanthemum (Chrysanthemum morifolium) - good for removing formaldehyde, benzene and ammonia from the air. Boston Fern (Nephrolepsis exaltata "Bostoniensis") - suggested to be the best for removing formaldehyde. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mrosla, Tina M Sent: Thursday, 22 September 2005 3:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] plants in histology lab We are trying to decide if we should have plants in our histology lab or not. It would be nice, for the formalin fumes and such, but we are wondering about contamination. Does anyone have any suggestions and would certain plants be better to have than others, or none at all? Thank you, Tina St. Joseph's Histology St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PKamalavenkatesh <@t> wockhardtin.com Mon Sep 26 00:18:45 2005 From: PKamalavenkatesh <@t> wockhardtin.com (PKamalavenkatesh@wockhardtin.com) Date: Mon Sep 26 00:15:26 2005 Subject: [Histonet] Ornithine Carbamyl Transferase in rat liver Message-ID: Dear All, I am wondering that i am not getting any feed back from histonetters regarding my posting on ornithine carbamyl transferase detection in rat liver. We need to standardize simple histochemical detection Ornithine Carbamyl Transferase in rat liver. We got some references which are mentioning a technique called "Mizutani technique for ornithine carbamoyltransferase detection in liver". If you are having any idea regarding this, we are in need of it. Regards kamalavenkatesh Wockhardt Research Center New Drug Discovery - Preclinical safety Evaluation India --------------------- Disclaimer ------------------------------------------------------------------------------ Information transmitted by this E-MAIL is proprietary to Wockhardt Ltd. and/ or its Group Companies and/or,its Customers and is intended for use only by the individual or entity to which it is addressed, and may contain information that is privileged, confidential or exempt from disclosure under applicable law. If you are not the intended recipient or it appears that this mail has been forwarded to you without proper authority, you are notified that any use or dissemination of this information in any manner is strictly prohibited. In such cases, please delete this mail from your records. --------------------------------------------------------------------------------------------------------------- From stevenk <@t> med.usyd.edu.au Mon Sep 26 00:43:51 2005 From: stevenk <@t> med.usyd.edu.au (stevenk) Date: Mon Sep 26 00:40:20 2005 Subject: [Histonet] Re:GFAP staining Message-ID: I was wondering if anyone can help with GFAP staining both at increasing the intensity and reducing the background. I was trying to stain astrocytes in crystat cut human brain sections, dissect them off the slide using PALM LCM system and then extract DNA from the resultant cells. So far the only fixative that allowed any staining was 5% Acetic Acid in 95%ETOH. That gave pale brown staining with about the same intensity background which makes distinguishing the cells difficult when not coverslipped. Sections were air dried and then fixed. Fixatives tried have been 30min and 16hr duration paraformaldehyde; 30min acetone; 10min 70% ethanol; 30min 5 acetic acid in 95% ethanol. The acetone fixed slides were air dried for an hour before applying the primary. The rest were washed in 0.1M Tris/saline and some were blocked with normal horse serum before applying GFAP. The positive control was formaldehyde fixed, paraffin embedded tissue which worked fine. the GFAP was rabbit anticow by DAKO. 1:400, 1:800, and 1:1500 dilution. Any assistance would be gratefully accepted. Stephen > -- Stephen Kum Jew Senior Technical Officer Neuropathology Department of Pathology Blackburn Building D06 University of Sydney NSW 2006 Australia Ph: 61 2 9351 6143 Fax: 61 2 9351 3429 From jkiernan <@t> uwo.ca Mon Sep 26 01:13:12 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Sep 26 01:13:37 2005 Subject: [Histonet] Ornithine Carbamyl Transferase in rat liver References: Message-ID: <43379178.4C6C008F@uwo.ca> If nobody has replied, it's probably because nobody understands your question. Volume 3 of the 4th edition of Pearse's Histochemistry (1991) includes discussion of an enzyme called ornithine carbamoyltransferase (EC 2.2.3.3). Is this the same as your "ornithine carbamyl transferase"? Pearse gives two lead-precipitation methods for ornithine carbamoyltransferase activity, on pages 577-578. Mituzani is cited (various mid-1960s papers) in Pearse's discussion of the techniques (pages 163-165), but Pearse's recommended methods are from 1969 and 1983 papers with different authors. Before doing anything in the lab, go to the library. Dig out Pearse vol 3 and check out all the references about the enzyme. There about 20; an afternon's work. Next, use Scopus or Web of Science to find recent publications that cite the major papers about histochemical localization of the enzyme. You will almost certainly encounter recent improvements to the method. John Kiernan Anatomy, UWO London, Canada. ----------------------- PKamalavenkatesh@wockhardtin.com wrote: > > Dear All, > > I am wondering that i am not getting any feed back from > histonetters regarding my posting on ornithine carbamyl transferase > detection in rat liver. We need to standardize simple histochemical > detection Ornithine Carbamyl Transferase in rat liver. We got some > references which are mentioning a technique called "Mizutani technique for > ornithine carbamoyltransferase detection in liver". If you are having any > idea regarding this, we are in need of it. > > Regards > kamalavenkatesh > Wockhardt Research Center > New Drug Discovery - Preclinical safety Evaluation > India > > --------------------- Disclaimer ------------------------------------------------------------------------------ > Information transmitted by this E-MAIL is proprietary to Wockhardt Ltd. and/ or its Group Companies > and/or,its Customers and is intended for use only by the individual or entity to which it is addressed, > and may contain information that is privileged, confidential or exempt from disclosure under applicable law. > If you are not the intended recipient or it appears that this mail has been forwarded to you without proper > authority, you are notified that any use or dissemination of this information in any manner is strictly > prohibited. In such cases, please delete this mail from your records. > --------------------------------------------------------------------------------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Sep 26 01:14:02 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Sep 26 01:14:14 2005 Subject: [Histonet] antigen retrieval for IHC References: <43347F7C.3070109@ski.org> Message-ID: <433791AA.6E4F2C17@uwo.ca> Maria, can you give chapter and verse for the Shi/Taylor and Puchrler/Meloan papers? Most papers with Shi and Taylor among the authors are about high temperature antigen retrieval (boiling water, with pH optima for various antigens). Holde Puchtler and Susan Meloan published many imortant papers about staining ad histochemistry in the 1970s=1980s. Susan M was a histonetter in the 1990s. ***************** Maria Mejia wrote: > > Hello, > > I believe it was sometime in June of this year that there was a > discussion on the > histonet regarding "the simplest form" of reversing the effects for formalin > fixation on tissue was to wash the tissue well (before) processing. > > Well, I just read the article "Antigen retrieval IHC: Past, Present, & > Future by > Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google this > article). It's > a very interesting! Anyway, the article has a section titled non-heating > AR method stating (this) same simple method by (Puchtler & Meloan). It also > goes on to say that Elias JM (1990) adopted this method routinely by storing > deparaffinized in fresh changes of 10% sucrose/PBS @ 4C overnight (before) > IHC. > > My questions are, does anyone know or tried this method used by Elias? > And, can anyone explain the mechanism of 10% sucrose/PBS play in this > AR method? > > Just curious! > > Maria Bartola Mejia > Smith-Kettlewell Eye Research Institute > San Francisco, CA 94115 > Email: maria@ski.org > Phone: (415)-345-2185 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bernaweston <@t> hotmail.com Mon Sep 26 06:21:08 2005 From: bernaweston <@t> hotmail.com (Bernadette Weston) Date: Mon Sep 26 06:21:25 2005 Subject: [Histonet] slide QC Message-ID: What method of QC checking slides(H&E) do you use, does the histologist check the slides microscopically before they take them to the pathologist or does the pathologist review them and give either a verbal or written evaluation of the day's work? If the histologist does the before check, how many slides are reviewed? Bernadette Weston HT Histology Supervisor Barberton Citizens Hospital Barberton,OH From mprice26 <@t> juno.com Mon Sep 26 08:13:34 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Mon Sep 26 08:16:19 2005 Subject: [Histonet] RE: Refurbished Histology Equipment Vendors Message-ID: <20050926.061415.296.602801@webmail33.nyc.untd.com> I am looking to buy a refurbished VIP processor. I have a VIP 2000 that is in good working condition that I would like to trade in on perhaps a newer model. I welcome all Vendors replies. Thank you. Marsha Price From cwscouten <@t> myneurolab.com Mon Sep 26 08:49:07 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Mon Sep 26 08:50:07 2005 Subject: [Histonet] Microwave PROCCESSING Message-ID: <5784D843593D874C93E9BADCB87342AB916663@tpiserver03.Coretech-holdings.com> Our position on "Microwave Processing" is as follows. Our position in the field comes from the fixation and processing of biological specimens for specimen preparation for transmission electron microscopy evaluation. There is no technology in the market place that is able to match our ability to prepare tissues to be examined ultrastructurally for electron microscopy evaluation. Our preservation of ultrastructure is equal to or better than traditional methods of specimen preparation for electron microscopy. Thus, we come to the field of histology looking at the preservation and processing of tissue for pathologic evaluation from a different aspect than any of the other microwave technologies present in the field of histology. The primary focus of our interest in histology is fixation using 10% neutral buffered formalin and bone decalcification using EDTA. We are able to give you fixation equal to a standard 24 hour fixation in 10% neutral buffered formalin in one hour. Fixation is the key to processing and all following results from routine H&E staining results to improved IHC results. Good fixation is the key to fast and excellent tissue processing. Specimens that are fixed using our microwave technology and protocol will allow the lab to process these microwave fixed specimens using a traditional tissue processor and chemicals using a short cycle processing protocol or perform rapid processing using a microwave processing schedule. We are able to decalcify bone specimens with EDTA in 1/10 the time it takes to decalcify under routine bone decalcification procedures. We are able to give you the quality results achieved using EDTA with the speed of formic acid protocols. Our microwave technology allows the lab to fix tissue in 1 hour. These tissues must be grossed to a thickness of 2 millimeters. 2 millimetes is the thickness that gives the best fixation of specimens in 24 hours and in our one hour fixation protocol. These microwave fixed specimens may then be processed on a routine tissue processor using standard reagents or processed rapidly in a microwave processing schedule. The H&E stain should equal or better a specimen processed on a routine fixation and precessing schedule and should result in improved IHC staining relults. Specimens greater than 2 millimeters will not achieve the quality fixation results using our microwave fixation protocol or using a routine 24 hour fixation protocol. Cordially, Lee Lee Dickey Product Marketing Manager McCormick Scientific 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 866-737-2540 Ledickey@mccormickscientific.com Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jesus Ellin Sent: Thursday, September 22, 2005 1:16 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave PROCCESSING Nice to see that everyone made it back from NSH and are doing well. I have a nice question that might start some serious discussion. Microwave proccessing!!!! My question is how many people out there are doing it and what issues did you have to work through to apply this technology to everyday use??? How are you handling the testing of your immuno's since you have to use tissue that has been run by the microwave proccessor??? How is the transcription being done??? What time are the pathologist coming in and how LATE are they staying?? Are you proccessing all tissue type or only small biopies, breast biopsies, etc.?? What road block have you come up against??? It this the rule to proccess all tissue or is this the exception?? How are you handling reference lab consultation with your tissue that is proccessed on the Microwave and sending it to a lab for instance AFIP for consult and they only use the conventional proccessing?? Is anyone out there doing both and how do they keep track of the different types of tissue that have been proccessed within the different proccessor, if you are indeed using both. What additional costs did you do you have, with there new product from the proccessor?? Our pathologist came back and are now foaming at the mouth for this technology. We are not against this, but there are avenues that need to be addressed, especially with IHC, FISH, and overall workflow. Any help would greatly be appreciated and needed at this time. Your in Formalin Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Mon Sep 26 08:50:16 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Mon Sep 26 08:50:46 2005 Subject: [Histonet] Free Ultra Microtome Message-ID: <6.1.1.1.2.20050926094853.019d30f0@mail.vet.upenn.edu> Denise Longworth Woodward Please contact me if you still want the microtome and let me know if you want to pick it up. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From gcallis <@t> montana.edu Mon Sep 26 09:44:17 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Sep 26 09:44:49 2005 Subject: [Histonet] RNAse decontamination in cryostats In-Reply-To: References: Message-ID: <6.0.0.22.1.20050926084101.01b699d0@gemini.msu.montana.edu> A friend who is an expert at LCM, dealing with collecting frozen sections for RNA work, wipes down the whole inside of the cryostat with 70% alcohol. You could make this up with RNAas away if you wanted to. She wipes down all surfaces, including under the sliding glass door/lid of the cryostat. If you wish I will put you in contact with her - she is not a Histonet subscriber, so you can discuss any fine tuning of cryostat RNase decontamination. 01:28 PM 9/23/2005, you wrote: >My question is similar to the current topic of discussion about >decontaminating cryostats, except that I need to remove any potential RNAse >activity. I have been cleaning the disposable blade and the blade holder >with 70% alcohol, which hasn't been shown to eliminate the pernicious little >buggers, but the commercial products such as RNAse away (Ambion) etc, are >water based and would freeze on the blade/blade holder which would make me >suspicious of their effectiveness. I am not able to continuously thaw the >cryostat and then use the products, as I would like to do daily. I am >wondering if anyone has any ideas or potential products that would work. >Thanks > >Patrick Laurie, HT (ASCP) >Neurogenomics Laboratory >Benaroya Research Institute >1201 9th Ave >Seattle, Wa 98101 >(206) 341-0681 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Traczyk7 <@t> aol.com Mon Sep 26 09:59:43 2005 From: Traczyk7 <@t> aol.com (Traczyk7@aol.com) Date: Mon Sep 26 10:00:03 2005 Subject: [Histonet] cryostat decontamination Message-ID: <1f0.44e1d1d4.306966df@aol.com> Greetings, I recently attended Gloria Limetti's seminar at NSH about cryostat and microtome features. She is from the University of Pitssburgh. Attendees were given an extensive spreadsheet listing which features were available on which models. In an effort not to come across as non vendor specific, Gloria assigned all the companies letters. Just looking over the features, it's fairly easy to decipher which company is which. I don't have my copy in front of me now but I believe there are 7 models with various types of decontamination systems offered. Hacker Instruments SL5000 is one of the units that is available with an automatic decontamination feature, perhaps Gloria will post the names of the other companies. As for UV decontamination, it is my understanding the the UV light is only effective on the surfaces of the cryostat and microtome chamber where the light actually comes into contact. Nooks and cranies would still need to be wiped down manually. As with Vinnie, I would be interested in reading a study or two on it's effectiveness in histology applications. Regards, Dorothy Murphy Traczyk National Sales Manager Hacker Instruments & Industries Inc. PO Box 1176 Winnsboro, SC 29180 1-800-442-2537 hackerlab@aol.com _www.hacker_ (http://www.hacker/) insruments.com From Jason.Thorsten <@t> us.crl.com Mon Sep 26 10:01:53 2005 From: Jason.Thorsten <@t> us.crl.com (Thorsten, Jason ) Date: Mon Sep 26 10:02:22 2005 Subject: [Histonet] unsubscribe Message-ID: Jason J. Thorsten, HT (ASCP) Charles River Laboratories Interventional and Surgical Services-Wisconsin Division 803 Prospect Avenue Osceola, Wisconsin 54020 715-294-4371, extension 258? 715-294-3723 fax jason.thorsten@us.crl.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Y. Wang Sent: Friday, July 29, 2005 3:08 PM To: Larry Woody Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] collagen precursors Larry, There are some abs against type I and type III procollagen available. There is a company called DiaSorin that has a radioimmunoassay kit for these too (human). They also may have the straight antibodies. I've never tied any of them but there are a few companies that have these abs (mainly for human). I believe Chemicon, mdbiosciences and novotec have them, to name a few. I hope this is of some help. Yak-Nam > Hi all: Does anyone out there know of any special stains or immuno > stains used for collagen precursors? Thanks Larry > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DCadoretteHall <@t> sprg.mercy.net Mon Sep 26 10:05:29 2005 From: DCadoretteHall <@t> sprg.mercy.net (Cadorette-Hall, Diane M) Date: Mon Sep 26 10:06:14 2005 Subject: [Histonet] Job Opening in Springfield Missouri Message-ID: <99648DDDA90EA647B3E8A1FD4AFD339801A2DA4D@sprgexelroy.sprg.mercy.net> I have a full time position available for a HT or HT eligible. Duties will be mainly in the gross room with rotation through other duty stations. Please email me for more details. Diane Cadorette-Hall, HT (ASCP) Pathology Laboratory Supervisor St. Johns Regional Health Center 1235 East Cherokee Springfield, Missouri 65804 (417) 820-6492 Fax: (417) 820-7790 DCadoretteHall@sprg.mercy.net From jbaez <@t> interscopepath.com Mon Sep 26 10:42:09 2005 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Mon Sep 26 10:42:45 2005 Subject: [Histonet] PA position Message-ID: <9E956D8FEB06C2408B08AC16498325E90139F8@adsl-67-113-77-28.dsl.lsan03.pacbell.net> We currently have a PA position available, great location in Southern California. Please e-mail for further details or fax resume. Janet Baez Histology Manager Interscope Pathology Canoga Park, Ca. Fax 818- 992-6654 From pruegg <@t> ihctech.net Mon Sep 26 10:58:46 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Sep 26 10:59:19 2005 Subject: [Histonet] antigen retrieval for IHC In-Reply-To: <433791AA.6E4F2C17@uwo.ca> Message-ID: <200509261558.j8QFwgAA013907@chip.viawest.net> Shi mentions in his book on pg.10 ANTIGEN RESTORATION "Simply bathing deparaffinized sections in a cold 20% sucrose-saline solution could, over several days, restore a certain amount of immunoreactivity." In Introduction to IHC by Polak and Van Noorden on pg 24 3.8.1 "The simplest form of reversing the effects of formalin is to wash the tissue well before processing" I am about to test these statements, as I just received tissues for an IHC project that have been in 10% NBF for over one year. I washed the tissues in running tap h20 for 2 hrs., I will now process them into paraffin. I am planning as well to put some of the sections in 20% sucrose at 4dc for 2 days if I have trouble getting good IHC signal. I will keep you all posted on this experiment. These are samples of mouse mammory tissue. I will be testing them with cleaved caspase 3 and Ki67 to start. I have both of these antibodies worked out very well for ffpe tissues fixed 24-48 hrs. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: Sunday, September 25, 2005 11:14 PM To: Maria Mejia Cc: pruegg@ihctech.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] antigen retrieval for IHC Maria, can you give chapter and verse for the Shi/Taylor and Puchrler/Meloan papers? Most papers with Shi and Taylor among the authors are about high temperature antigen retrieval (boiling water, with pH optima for various antigens). Holde Puchtler and Susan Meloan published many imortant papers about staining ad histochemistry in the 1970s=1980s. Susan M was a histonetter in the 1990s. ***************** Maria Mejia wrote: > > Hello, > > I believe it was sometime in June of this year that there was a > discussion on the histonet regarding "the simplest form" of reversing > the effects for formalin fixation on tissue was to wash the tissue > well (before) processing. > > Well, I just read the article "Antigen retrieval IHC: Past, Present, & > Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google > this article). It's a very interesting! Anyway, the article has a > section titled non-heating AR method stating (this) same simple method > by (Puchtler & Meloan). It also goes on to say that Elias JM (1990) > adopted this method routinely by storing deparaffinized in fresh > changes of 10% sucrose/PBS @ 4C overnight (before) IHC. > > My questions are, does anyone know or tried this method used by Elias? > And, can anyone explain the mechanism of 10% sucrose/PBS play in this > AR method? > > Just curious! > > Maria Bartola Mejia > Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 > Email: maria@ski.org > Phone: (415)-345-2185 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From maria <@t> ski.org Mon Sep 26 11:31:15 2005 From: maria <@t> ski.org (Maria Mejia) Date: Mon Sep 26 11:31:56 2005 Subject: [Histonet] antigen retrieval for IHC References: <43347F7C.3070109@ski.org> <433791AA.6E4F2C17@uwo.ca> Message-ID: <43382253.3070909@ski.org> Hello Everyone, Here's the information: "Antigen Retrieval IHC: Past, Present & Future" by Shan-Rong Shi, Richard J. Cote & Clive R. Taylor is in the Journal of Histochemistry & Cytochemistry, Vol. 45, 327-344, 1997 Puchtler H., Meloan S.N. (1985) "On the chemistry of formaldehyde fixation & its effects on IHC reactions. Histochemistry 82: 201-204 Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 John Kiernan wrote: >Maria, can you give chapter and verse for the Shi/Taylor >and Puchrler/Meloan papers? Most papers with Shi and Taylor >among the authors are about high temperature antigen retrieval >(boiling water, with pH optima for various antigens). > >Holde Puchtler and Susan Meloan published many imortant papers >about staining ad histochemistry in the 1970s=1980s. Susan M >was a histonetter in the 1990s. > >***************** > >Maria Mejia wrote: > > >>Hello, >> >>I believe it was sometime in June of this year that there was a >>discussion on the >>histonet regarding "the simplest form" of reversing the effects for formalin >>fixation on tissue was to wash the tissue well (before) processing. >> >>Well, I just read the article "Antigen retrieval IHC: Past, Present, & >>Future by >>Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google this >>article). It's >>a very interesting! Anyway, the article has a section titled non-heating >>AR method stating (this) same simple method by (Puchtler & Meloan). It also >>goes on to say that Elias JM (1990) adopted this method routinely by storing >>deparaffinized in fresh changes of 10% sucrose/PBS @ 4C overnight (before) >>IHC. >> >>My questions are, does anyone know or tried this method used by Elias? >>And, can anyone explain the mechanism of 10% sucrose/PBS play in this >>AR method? >> >>Just curious! >> >>Maria Bartola Mejia >>Smith-Kettlewell Eye Research Institute >>San Francisco, CA 94115 >>Email: maria@ski.org >>Phone: (415)-345-2185 >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> From m.wei <@t> biogenex.com Mon Sep 26 12:05:15 2005 From: m.wei <@t> biogenex.com (May Wei) Date: Mon Sep 26 12:05:53 2005 Subject: [Histonet] antigen retrieval for IHC Message-ID: <37DC9F93CF7F864182D0463EF93D571B0B5043@ISLETON2.california.biogenex.com> One of papers with Shi/Taylor is "Standardization of Immunohistochemistry Based on Antigen Retrieval Technique for Routine Formalin-fixed Tissue Sections" in Applied Immunohistochemistry 6(2): 89-96, 1998. I have the article. I also have the list of AR paper references. If you like to receive it, I can send to you. May Wei -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Sunday, September 25, 2005 11:14 PM To: Maria Mejia Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] antigen retrieval for IHC Maria, can you give chapter and verse for the Shi/Taylor and Puchrler/Meloan papers? Most papers with Shi and Taylor among the authors are about high temperature antigen retrieval (boiling water, with pH optima for various antigens). Holde Puchtler and Susan Meloan published many imortant papers about staining ad histochemistry in the 1970s=1980s. Susan M was a histonetter in the 1990s. ***************** Maria Mejia wrote: > > Hello, > > I believe it was sometime in June of this year that there was a > discussion on the histonet regarding "the simplest form" of reversing > the effects for formalin fixation on tissue was to wash the tissue > well (before) processing. > > Well, I just read the article "Antigen retrieval IHC: Past, Present, & > Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google > this article). It's a very interesting! Anyway, the article has a > section titled non-heating AR method stating (this) same simple method > by (Puchtler & Meloan). It also goes on to say that Elias JM (1990) > adopted this method routinely by storing deparaffinized in fresh > changes of 10% sucrose/PBS @ 4C overnight (before) IHC. > > My questions are, does anyone know or tried this method used by Elias? > And, can anyone explain the mechanism of 10% sucrose/PBS play in this > AR method? > > Just curious! > > Maria Bartola Mejia > Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 > Email: maria@ski.org > Phone: (415)-345-2185 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Mon Sep 26 12:12:32 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Mon Sep 26 12:12:53 2005 Subject: [Histonet] antigen retrieval for IHC Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D178@usca0082k08.labvision.apogent.com> It's true that most papers by Shi et al are on high-temperature antigen retrieval, but in their book on antigen retrieval (Antigen retrieval Techniques, Immunohistochemistry and Molecular Morphology, ed. Shan-Rong Shi et al, Eaton Publishing, 2000) they make clear that the term "antigen retrieval" should cover all types -enzyme, heat, or other. In fact they make a plea for everyone publishing methods to use the term "antigen retrieval" so matter what the method so that databases are easy to search. With the plethora of terms used to decribe the methods it is nearly impossible to find all the methods developed in the past 14 years. Part of the reason for people using all the different terms was the assumption by many that BioGenex had trademarked the term. BioGenex did apply for a trademark, but then withdrew it. So the term is open to all to use. However, BioGenex does have a patent on a very specific method of antigen retrieval, and some antigen retrieval buffer formulations. Tim Morken Lab Vision - Neomarkers www.labvision.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Sunday, September 25, 2005 11:14 PM To: Maria Mejia Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] antigen retrieval for IHC Maria, can you give chapter and verse for the Shi/Taylor and Puchrler/Meloan papers? Most papers with Shi and Taylor among the authors are about high temperature antigen retrieval (boiling water, with pH optima for various antigens). Holde Puchtler and Susan Meloan published many imortant papers about staining ad histochemistry in the 1970s=1980s. Susan M was a histonetter in the 1990s. ***************** Maria Mejia wrote: > > Hello, > > I believe it was sometime in June of this year that there was a > discussion on the histonet regarding "the simplest form" of reversing > the effects for formalin fixation on tissue was to wash the tissue > well (before) processing. > > Well, I just read the article "Antigen retrieval IHC: Past, Present, & > Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google > this article). It's > a very interesting! Anyway, the article has a section titled non-heating > AR method stating (this) same simple method by (Puchtler & Meloan). It also > goes on to say that Elias JM (1990) adopted this method routinely by storing > deparaffinized in fresh changes of 10% sucrose/PBS @ 4C overnight (before) > IHC. > > My questions are, does anyone know or tried this method used by Elias? > And, can anyone explain the mechanism of 10% sucrose/PBS play in this > AR method? > > Just curious! > > Maria Bartola Mejia > Smith-Kettlewell Eye Research Institute > San Francisco, CA 94115 > Email: maria@ski.org > Phone: (415)-345-2185 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Melissa.Gonzalez <@t> cellgenesys.com Mon Sep 26 12:50:34 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Mon Sep 26 12:51:54 2005 Subject: [Histonet] Fluorescent Illumination Systems: Suggestions please? Message-ID: Hello all, Sorry to re-post, but I only received one response to the following: I am currently in the market to upgrade our existing HB0 100 lamp house, which is very old, to several of new systems. Either the newest version of the HB0 100 from Zeiss, a mercury halide system called X-Cite from EXFO, or a xenon based burner called Lambda LS from Sutter Instruments. They are all comparable in price, but my main concern is increasing the brightness of my signal and evenly illuminating the field of view. I mostly look at DAPI, eGFP, FITC, and Alexa 594, which from spectral outputs, each one of these systems appears to have a different weakness in one of the above. On a plus, the Xcite rates their bulbs at 1500 hours, and the Lambda at 500-1000hrs. Does anyone out there have any input/comments/experience with these systems? (This would be hooked up to a Zeiss Axioplan) Please let me know if you have any info, as I have been unsuccessful so far with demos and need to place an order soon. Thanks a lot -I really appreciate it, Melissa From info <@t> instrumedics.com Mon Sep 26 12:53:16 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Mon Sep 26 12:53:47 2005 Subject: [Histonet] cryostat decontamination References: <1f0.44e1d1d4.306966df@aol.com> Message-ID: <01d401c5c2c3$336ba0f0$6401a8c0@INSTRUMEDICS22> Instrumedics' Cryo-Vac-Away keeps the cryostat free of trimming debris. The debris is suctioned away as it is generated at the block face when you trim the block. The debris is captured in a primary filter that is inside the cryostat and downstream of the primary filter is a viral/ bacterial filter that captures pathogens. The cryostat remains spotless and substantially reduces the hazards associated with frozen sectioning. Please visit www.instrumedics.com for details. Bernice ----- Original Message ----- From: To: Sent: Monday, September 26, 2005 10:59 AM Subject: Re: [Histonet] cryostat decontamination > Greetings, > I recently attended Gloria Limetti's seminar at NSH about cryostat and > microtome features. She is from the University of Pitssburgh. Attendees > were > given an extensive spreadsheet listing which features were available on > which > models. In an effort not to come across as non vendor specific, Gloria > assigned all the companies letters. Just looking over the features, it's > fairly > easy to decipher which company is which. I don't have my copy in front > of me > now but I believe there are 7 models with various types of > decontamination > systems offered. > Hacker Instruments SL5000 is one of the units that is available with an > automatic decontamination feature, perhaps Gloria will post the names of > the > other companies. > As for UV decontamination, it is my understanding the the UV light is only > effective on the surfaces of the cryostat and microtome chamber where the > light > actually comes into contact. Nooks and cranies would still need to be > wiped down manually. As with Vinnie, I would be interested in reading a > study or > two on it's effectiveness in histology applications. > Regards, > > Dorothy Murphy Traczyk > National Sales Manager > Hacker Instruments & Industries Inc. > PO Box 1176 > Winnsboro, SC 29180 > 1-800-442-2537 > hackerlab@aol.com > _www.hacker_ (http://www.hacker/) insruments.com > > From pmarcum <@t> vet.upenn.edu Mon Sep 26 13:01:50 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Mon Sep 26 13:02:23 2005 Subject: [Histonet] antigen retrieval for IHC In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D178@usca0082k08.labvi sion.apogent.com> References: <0556BE8AC5551E4E8AF6BB9E42509BA20328D178@usca0082k08.labvision.apogent.com> Message-ID: <6.1.1.1.2.20050926140018.01a53ce0@mail.vet.upenn.edu> Thanks for the update Tim. I know for years we have all been careful about using antigen retrieval due to the BioGenex issue. They do have a patent on the one tyoe and it was spread to all for a while. Pam Marcum At 01:12 PM 9/26/2005, Morken, Tim - Labvision wrote: >It's true that most papers by Shi et al are on high-temperature antigen >retrieval, but in their book on antigen retrieval (Antigen retrieval >Techniques, Immunohistochemistry and Molecular Morphology, ed. Shan-Rong Shi >et al, Eaton Publishing, 2000) they make clear that the term "antigen >retrieval" should cover all types -enzyme, heat, or other. In fact they make >a plea for everyone publishing methods to use the term "antigen retrieval" >so matter what the method so that databases are easy to search. With the >plethora of terms used to decribe the methods it is nearly impossible to >find all the methods developed in the past 14 years. Part of the reason for >people using all the different terms was the assumption by many that >BioGenex had trademarked the term. BioGenex did apply for a trademark, but >then withdrew it. So the term is open to all to use. However, BioGenex does >have a patent on a very specific method of antigen retrieval, and some >antigen retrieval buffer formulations. > >Tim Morken >Lab Vision - Neomarkers >www.labvision.com > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan >Sent: Sunday, September 25, 2005 11:14 PM >To: Maria Mejia >Cc: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] antigen retrieval for IHC > > >Maria, can you give chapter and verse for the Shi/Taylor >and Puchrler/Meloan papers? Most papers with Shi and Taylor among the >authors are about high temperature antigen retrieval (boiling water, with pH >optima for various antigens). > >Holde Puchtler and Susan Meloan published many imortant papers about >staining ad histochemistry in the 1970s=1980s. Susan M >was a histonetter in the 1990s. > >***************** > >Maria Mejia wrote: > > > > Hello, > > > > I believe it was sometime in June of this year that there was a > > discussion on the histonet regarding "the simplest form" of reversing > > the effects for formalin fixation on tissue was to wash the tissue > > well (before) processing. > > > > Well, I just read the article "Antigen retrieval IHC: Past, Present, & > > Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google > > this article). It's > > a very interesting! Anyway, the article has a section titled non-heating > > AR method stating (this) same simple method by (Puchtler & Meloan). It >also > > goes on to say that Elias JM (1990) adopted this method routinely by >storing > > deparaffinized in fresh changes of 10% sucrose/PBS @ 4C overnight (before) > > IHC. > > > > My questions are, does anyone know or tried this method used by Elias? > > And, can anyone explain the mechanism of 10% sucrose/PBS play in this > > AR method? > > > > Just curious! > > > > Maria Bartola Mejia > > Smith-Kettlewell Eye Research Institute > > San Francisco, CA 94115 > > Email: maria@ski.org > > Phone: (415)-345-2185 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From kmerriam2003 <@t> yahoo.com Mon Sep 26 13:34:06 2005 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Mon Sep 26 13:34:25 2005 Subject: [Histonet] Low temp Antigen Retrieval for Bone Sections Message-ID: <20050926183406.77615.qmail@web50312.mail.yahoo.com> Hello All, I will be performing some IHC on mouse femurs (about 12 or so different antibodies). In the past, I have had a lot of trouble keeping these sections on the slides during HIER. I did a quick search on the archives and there were lots of suggestions, but nothing definitive. I was reading an article about "low temp AR", overnight at 60C in TEG buffer, pH 9.0. Has anyone tried this method? Also - what does TEG buffer stand for? Any help would be greatly appreciated. Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From kgibbon <@t> qltinc.com Mon Sep 26 14:22:30 2005 From: kgibbon <@t> qltinc.com (kgibbon@qltinc.com) Date: Mon Sep 26 14:23:03 2005 Subject: [Histonet] Fluorescent Illumination Systems: Suggestions please? Message-ID: Hi Melissa, I have used the Sutter xenon Lambda unit for a few years now and really like it, The spectrum is much more even than a standard mercury bulb. I have the model with the extended bandwidth so I could use it well into the UV wavelengths but it does produce ozone which must be vented, I believe they have another model that does not do this but has a higher minimum wavelength. Your choice, depending what fluorochromes you want to light up. Kevin Gibbon QLT Inc. |---------+-------------------------------------------> | | "Melissa Gonzalez" | | | | | | Sent by: | | | | | | | | | | | | 09/26/2005 10:50 AM | | | | |---------+-------------------------------------------> >------------------------------------------------------------------------------------------------------------------------------| | | | To: | | cc: | | Subject: [Histonet] Fluorescent Illumination Systems: Suggestions please? | >------------------------------------------------------------------------------------------------------------------------------| Hello all, Sorry to re-post, but I only received one response to the following: I am currently in the market to upgrade our existing HB0 100 lamp house, which is very old, to several of new systems. Either the newest version of the HB0 100 from Zeiss, a mercury halide system called X-Cite from EXFO, or a xenon based burner called Lambda LS from Sutter Instruments. They are all comparable in price, but my main concern is increasing the brightness of my signal and evenly illuminating the field of view. I mostly look at DAPI, eGFP, FITC, and Alexa 594, which from spectral outputs, each one of these systems appears to have a different weakness in one of the above. On a plus, the Xcite rates their bulbs at 1500 hours, and the Lambda at 500-1000hrs. Does anyone out there have any input/comments/experience with these systems? (This would be hooked up to a Zeiss Axioplan) Please let me know if you have any info, as I have been unsuccessful so far with demos and need to place an order soon. Thanks a lot -I really appreciate it, Melissa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This electronic transmission (including any and all attachments) is intended solely for the use of the individual or entity to whom it is addressed and may contain information that is privileged and/or confidential. If you are not the intended recipient of this electronic transmission, you are hereby notified that any disclosure, copying or distribution, or the taking of any action in reliance upon the contents of this electronic transmission, is strictly prohibited, and you are further requested to purge this electronic transmission and all copies thereof from your computer system. From PMonfils <@t> Lifespan.org Mon Sep 26 14:59:28 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Sep 26 14:59:55 2005 Subject: [Histonet] Low temp Antigen Retrieval for Bone Sections Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175E3@lsexch.lsmaster.lifespan.org> I haven't tried this method of antigen retrieval, but TEG buffer is Tris-EGTA buffer. The usual formula (I have seen a couple of variations) is: Distilled water 1,000 ml Trisma base 1.21 gram EGTA 0.19 gram The usual target pH is between 8.95 and 9.0 EGTA is ethylene glycol bis(B-aminoethyl)-N,N,N',N' tetraacetic acid. (The capital "B" in the above name represents "beta". My email program doesn't support the symbol.) > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kim > Merriam > Sent: Monday, September 26, 2005 11:34 AM > To: Histonet > Subject: [Histonet] Low temp Antigen Retrieval for Bone Sections > > Hello All, > > I will be performing some IHC on mouse femurs (about 12 or so different > antibodies). In the past, I have had a lot of trouble keeping these > sections on the slides during HIER. I did a quick search on the archives > and there were lots of suggestions, but nothing definitive. > > I was reading an article about "low temp AR", overnight at 60C in TEG > buffer, pH 9.0. Has anyone tried this method? Also - what does TEG buffer > stand for? > > Any help would be greatly appreciated. > > Kim > > > Kim Merriam > Novartis > Cambridge, MA > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Barb.Richmond <@t> mckennan.org Mon Sep 26 15:29:00 2005 From: Barb.Richmond <@t> mckennan.org (Barb Richmond) Date: Mon Sep 26 15:29:16 2005 Subject: [Histonet] cpt codes for direct smears Message-ID: I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? From bhewlett <@t> cogeco.ca Mon Sep 26 15:31:28 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Mon Sep 26 15:31:56 2005 Subject: [Histonet] antigen retrieval for IHC References: <200509261558.j8QFwgAA013907@chip.viawest.net> Message-ID: <002101c5c2d9$467fce80$6400a8c0@mainbox> Patsy, You will need to wash MUCH longer than 2 hours in running tap water in order to obtain significant reversal after I year in NBF. The Helander paper quoted in my NSH handout gives about 6 days to effect a 90% reversal and that was following 24 hour fixation. Best regards, Bryan ----- Original Message ----- From: "Patsy Ruegg" To: ; "'Maria Mejia'" Cc: Sent: Monday, September 26, 2005 11:58 AM Subject: RE: [Histonet] antigen retrieval for IHC > Shi mentions in his book on pg.10 ANTIGEN RESTORATION "Simply > bathing deparaffinized sections in a cold 20% sucrose-saline solution > could, > over several days, restore a certain amount of immunoreactivity." > > In Introduction to IHC by Polak and Van Noorden on pg 24 3.8.1 "The > simplest > form of reversing the effects of formalin is to wash the tissue well > before > processing" > > I am about to test these statements, as I just received tissues for an IHC > project that have been in 10% NBF for over one year. I washed the tissues > in running tap h20 for 2 hrs., I will now process them into paraffin. I > am > planning as well to put some of the sections in 20% sucrose at 4dc for 2 > days if I have trouble getting good IHC signal. I will keep you all > posted > on this experiment. These are samples of mouse mammory tissue. I will be > testing them with cleaved caspase 3 and Ki67 to start. I have both of > these > antibodies worked out very well for ffpe tissues fixed 24-48 hrs. > > Patsy > > > Patsy Ruegg, HT(ASCP)QIHC > IHCtech, LLC > Fitzsimmons BioScience Park > 12635 Montview Blvd. Suite 216 > Aurora, CO 80010 > P-720-859-4060 > F-720-859-4110 > wk email pruegg@ihctech.net > web site www.ihctech.net > > > This email is confidential and intended solely for the use of the > Person(s) > ('the intended recipient') to whom it was addressed. Any views or opinions > presented are solely those of the author. It may contain information that > is > privileged & confidential within the meaning of applicable law. > Accordingly > any dissemination, distribution, copying, or other use of this message, or > any of its contents, by any person other than the intended recipient may > constitute a breach of civil or criminal law and is strictly prohibited. > If > you are NOT the intended recipient please contact the sender and dispose > of > this e-mail as soon as possible. > > > -----Original Message----- > From: John Kiernan [mailto:jkiernan@uwo.ca] > Sent: Sunday, September 25, 2005 11:14 PM > To: Maria Mejia > Cc: pruegg@ihctech.net; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] antigen retrieval for IHC > > Maria, can you give chapter and verse for the Shi/Taylor and > Puchrler/Meloan > papers? Most papers with Shi and Taylor among the authors are about high > temperature antigen retrieval (boiling water, with pH optima for various > antigens). > > Holde Puchtler and Susan Meloan published many imortant papers about > staining ad histochemistry in the 1970s=1980s. Susan M was a histonetter > in > the 1990s. > > ***************** > > Maria Mejia wrote: >> >> Hello, >> >> I believe it was sometime in June of this year that there was a >> discussion on the histonet regarding "the simplest form" of reversing >> the effects for formalin fixation on tissue was to wash the tissue >> well (before) processing. >> >> Well, I just read the article "Antigen retrieval IHC: Past, Present, & >> Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google >> this article). It's a very interesting! Anyway, the article has a >> section titled non-heating AR method stating (this) same simple method >> by (Puchtler & Meloan). It also goes on to say that Elias JM (1990) >> adopted this method routinely by storing deparaffinized in fresh >> changes of 10% sucrose/PBS @ 4C overnight (before) IHC. >> >> My questions are, does anyone know or tried this method used by Elias? >> And, can anyone explain the mechanism of 10% sucrose/PBS play in this >> AR method? >> >> Just curious! >> >> Maria Bartola Mejia >> Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 >> Email: maria@ski.org >> Phone: (415)-345-2185 >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gcallis <@t> montana.edu Mon Sep 26 16:04:48 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Sep 26 16:05:07 2005 Subject: slides and RE: [Histonet] Low temp Antigen Retrieval for Bone Sections In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE017175E3@lsexch.lsmaster.l ifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE017175E3@lsexch.lsmaster.lifespan.org> Message-ID: <6.0.0.22.1.20050926150104.01b40890@gemini.msu.montana.edu> I strongly suggest if you try this retrieval, that you use Erie Scientific EXCEL slides, which are coated specially to reduce the effects of chelator based retrieval solutions at high aka alkaline pH. Many people experience section loss with this pH and chelator, so Erie developed a new positively charged coating to counteract losing precious sections. You can access samples from them to see if they work out for you, contact your Erie sales rep or even Erie directly, they have a website. Biogenex also has a decalcified bone retrieval solution, another one to try out. At 01:59 PM 9/26/2005, you wrote: >I haven't tried this method of antigen retrieval, but TEG buffer is >Tris-EGTA buffer. The usual formula (I have seen a couple of variations) is: > >Distilled water 1,000 ml >Trisma base 1.21 gram >EGTA 0.19 gram > >The usual target pH is between 8.95 and 9.0 > >EGTA is ethylene glycol bis(B-aminoethyl)-N,N,N',N' tetraacetic acid. > >(The capital "B" in the above name represents "beta". My email program >doesn't support the symbol.) > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kim > > Merriam > > Sent: Monday, September 26, 2005 11:34 AM > > To: Histonet > > Subject: [Histonet] Low temp Antigen Retrieval for Bone Sections > > > > Hello All, > > > > I will be performing some IHC on mouse femurs (about 12 or so different > > antibodies). In the past, I have had a lot of trouble keeping these > > sections on the slides during HIER. I did a quick search on the archives > > and there were lots of suggestions, but nothing definitive. > > > > I was reading an article about "low temp AR", overnight at 60C in TEG > > buffer, pH 9.0. Has anyone tried this method? Also - what does TEG buffer > > stand for? > > > > Any help would be greatly appreciated. > > > > Kim > > > > > > Kim Merriam > > Novartis > > Cambridge, MA > > __________________________________________________ > > Do You Yahoo!? > > Tired of spam? Yahoo! Mail has the best spam protection around > > http://mail.yahoo.com > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From jbaez <@t> interscopepath.com Mon Sep 26 16:08:53 2005 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Mon Sep 26 16:09:19 2005 Subject: [Histonet] slide QC Message-ID: <9E956D8FEB06C2408B08AC16498325E90139FA@adsl-67-113-77-28.dsl.lsan03.pacbell.net> We run one slide of an appendix everyday and date the slide. The histologist evaluates the slide before running the rest of the workload. The Pathologists then documents the quality on our QC sheet. -----Original Message----- From: Bernadette Weston [mailto:bernaweston@hotmail.com] Sent: Monday, September 26, 2005 4:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] slide QC What method of QC checking slides(H&E) do you use, does the histologist check the slides microscopically before they take them to the pathologist or does the pathologist review them and give either a verbal or written evaluation of the day's work? If the histologist does the before check, how many slides are reviewed? Bernadette Weston HT Histology Supervisor Barberton Citizens Hospital Barberton,OH _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jan.Minshew <@t> leica-microsystems.com Mon Sep 26 17:14:38 2005 From: Jan.Minshew <@t> leica-microsystems.com (Jan.Minshew@leica-microsystems.com) Date: Mon Sep 26 17:14:48 2005 Subject: [Histonet] Cryostat decontamination Message-ID: Hi Dorothy and Histonet subscribers, I'm writing in response to comments I've seen in the past few days about UV disinfection procedures for cryostats. I'm not sure if the listing on the spreadsheet from Gloria's workshop referred to our instrument or not, but Leica recently released the CM1850 UV cryostat with ultraviolet light (UVC) disinfection...so I thought I should chime in. We had the same questions about cryostat disinfection that many of you have had. Therefore, as part of our development process we hired an independent laboratory to perform tests that would verify the efficacy of UVC exposure against specific pathogens in the CM1850 at cold temperatures. The results assured us that our system is a convenient and safe means of inactivating microorganisms in the air and on exposed surfaces at temperatures down to -20?C and that using the system significantly reduces the risk of infection to the operator. We proudly provide a CD that contains the certificate from the consultant, details of how the tests were performed and the results that were obtained. Whether a cryostat has a built-in disinfection system of any kind or not, there are several very important things to remember about disinfecting cryostats. 1. Before beginning a disinfection protocol, don personal protective equipment (puncture and penetration resistant gloves, gowns, etc). 2. Remove all debris from the cryostat and disposed of it according to the policies and procedures of your institution. The debris must be removed because organic material (blood and proteins) may contain high concentrations of microorganisms and could possibly inactivate chemical germicides or prevent access to contaminated surfaces. 3. Use 70% ethyl alcohol to clean the cryostat. The germicidal activity of ethyl alcohol is most effective in that range and it has an advantage over isopropyl alcohol of being able to kill hydrophilic viruses. 4. For those of us in the USA (other countries have access to other products), the EPA maintains a list of Antimicrobial Chemical/Registration Number Indexes and it is posted on their website http://www.epa.gov/oppad001/chemregindex.htm. From this link you can find agents effective against bloodborne pathogens such as Mycobacterium tuberculosis, human HIV-1 virus, Hepatitis B or Hepatitis C virus. It is critical to remember that NONE of these solutions have been tested at low temperatures and they can only be used at room temperature. 5. Do not create aerosols by spraying disinfectant (or anything else) in an open cryostat chamber. Pour disinfectants onto absorbent disposable towels and allow them to remain in contact with contaminated surfaces for the length of time specified in the instructions of the individual agents. I hope this information is useful. Please let me know if you have any questions. Best wishes to all, Jan Minshew HT, HTL(ASCP) Marketing Manager Leica Microsystems, Inc. 2345 Waukegan Rd Bannockburn, IL 60015 800.248.0123 x7051 Traczyk7@aol.com Sent by: To: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: (bcc: Jan Minshew/USDER/West/Leica) western.edu Subject: Re: [Histonet] cryostat decontamination 09/26/2005 09:59 AM Greetings, I recently attended Gloria Limetti's seminar at NSH about cryostat and microtome features. She is from the University of Pitssburgh. Attendees were given an extensive spreadsheet listing which features were available on which models. In an effort not to come across as non vendor specific, Gloria assigned all the companies letters. Just looking over the features, it's fairly easy to decipher which company is which. I don't have my copy in front of me now but I believe there are 7 models with various types of decontamination systems offered. Hacker Instruments SL5000 is one of the units that is available with an automatic decontamination feature, perhaps Gloria will post the names of the other companies. As for UV decontamination, it is my understanding the the UV light is only effective on the surfaces of the cryostat and microtome chamber where the light actually comes into contact. Nooks and cranies would still need to be wiped down manually. As with Vinnie, I would be interested in reading a study or two on it's effectiveness in histology applications. Regards, Dorothy Murphy Traczyk National Sales Manager Hacker Instruments & Industries Inc. PO Box 1176 Winnsboro, SC 29180 1-800-442-2537 hackerlab@aol.com _www.hacker_ (http://www.hacker/) insruments.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From Sandy.Nelson <@t> tenethealth.com Mon Sep 26 18:03:15 2005 From: Sandy.Nelson <@t> tenethealth.com (Nelson, Sandy - PPH) Date: Mon Sep 26 18:03:52 2005 Subject: [Histonet] RE: Posting for Histology List Serve Message-ID: > Subject: Histology Management Position > > A 500 bed hospital adjacent to the Houston Medical Center has an opening for a full time Histology Supervisor. This candidate should have strong leadership skills and prior management experience is preferred. Interested? Call 713.527.5292. > > Sandra Nelson > Director Laboratory Services > Park Plaza Hospital > Houston, Texas > 713.527.5292 > sandy.nelson@tenethealth.com > ** This is not intended to be a legally binding or legally effective signature > > > From anh2006 <@t> med.cornell.edu Mon Sep 26 19:27:30 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Mon Sep 26 19:23:23 2005 Subject: [Histonet] Staining fibrin - thrombus Message-ID: Hi All! If you wanted to stain a paraffin section (frozen also available) of mouse tissue and prove what you are seeing is a result of a thrombotic event what stains would you use? I was thinking to do an anti-fibrin immunostain (antibody suggestions welcome) and perhaps some sort of fibrin special stain (I came up with Carstairs' and Weigert's as probably being the best). Any recommendations for antibodies, techniques, protocols, words of wisdom? Please any information, detailed or not, will be put to good use ... and thanks! Andrea -- From katri <@t> cogeco.ca Mon Sep 26 20:09:38 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Mon Sep 26 20:09:49 2005 Subject: [Histonet] IHC background problems on chicken embryos Message-ID: <000b01c5c300$225661f0$6a9a9618@Katri> Hi Histonetters, A colleague of mine is trying to get VWF (Dakocytomation) and VEGF(R&D Systems) working on chicken tissue. The antibodies are performing well on human and rat tissue, but excessive background is the problem with the chicken. Would the blocking serums (normal goat and normal rabbit respectively) be the problem as well as the biotinylated secondary reagents raised in goat and rabbit? My experience is strictly with human tissues, so I am stumped. To my knowledge neither of these antibodies have been proven to work with chicken, so maybe that is the problem. Any advice from "chicken" people? Katri Katri Tuomala Hamilton, Ontario, Canada From ralphpu <@t> alleninstitute.org Mon Sep 26 20:29:53 2005 From: ralphpu <@t> alleninstitute.org (Ralph Puchalski) Date: Mon Sep 26 20:30:13 2005 Subject: [Histonet] Pink precipitate is monoformazan from BCIP/NBT reaction on ISH? Message-ID: I am trying to figure out the origin of the pink precipitate in the image I posted on http://www.histonet.org/site_images_frame.asp Please go to the image entitled: Pink Precipitate ver2.jpg. It is at the top of the list on 9/26/05, posted by Ralph Puchalski. To see the pink precipitate artifact, please open the image and set the scroll bars on the bottom and right side of the image at their 1/2 way points. It is ugly! I think this precipitate is the aggregation of the monoformazan intermediate generated from NBT (after dephosphorylation of BCIP by alkaline phosphatase) that is not completely reduced to diformazan, the insoluble dark blue or black precipitate that labels cells expressing target mRNAs in our in situ hybridization reactions. We don't know how the monoformazan adheres to the sections of tissue. It appears to be non-covalent due to the tendency of the monoformazan to migrate under the coverslip in the aqueous mounting medium that has yet to dry, and form clumps or aggregates or pools as seen in the picture. The monoformazan is soluble in ethanol so doesn't form pools or aggregates. But as soon as it is exposed to an aqueous medium, it precipitates. If we mount the post-ISH tissue sections with an organic based mounting medium like UV-CureMount (Instrumedics), the monoformazan might cause the entire section to turn to a brown tint as seen on http://www.histonet.org/site_images_frame.asp Please go to the image Brown Tint 1.jpg. It is 4th image from the top on 9/26, posted by Ralph Puchalski. The lower image is mounted with UV CureMount, and the upper was with aqueous Hydro Matrix. There is no pooling or precipitation of monoformazan with UV CureMount, but I think it does cause the entire section to turn brown. Question: How do we eliminate this problem, which I believe is monoformazan? If we reduce it fully using ascorbate, the section (later mounted with HyrdoMatrix) turns dark blue or purple, the same color as our ISH signal. We have tried washing off the monoformazan with 100% ethanol prior to coverslipping, but only small amounts are removed. 100% acetone also does not work effectively. Please let me know if you have any ideas that might help us eliminate this problem. Thank you, Ralph Ralph Puchalski, Ph.D. Manager, Process Engineering and Automation Allen Institute for Brain Science 551 N. 34th Street, Suite 200 Seattle, WA 98103 ralphpu@alleninstitute.org Tel: 206-548-7041 Fax: 206-548-7071 www.brainatlas.org From histology <@t> 2hosts.com Mon Sep 26 20:28:56 2005 From: histology <@t> 2hosts.com (Histology) Date: Mon Sep 26 20:30:48 2005 Subject: [Histonet] Microwave PROCCESSING References: Message-ID: <009f01c5c302$d761a9e0$c901fec0@FAMILYROOM> We are using microwave processing. We've had to process fatty tissues, lymph nodes and cell blocks on a standard VIP, but are using the Sakura Xpress for the rest. We've recently had a software and reagent update which will allow all tissues types to run on the standard 1.5 hour program. There was some adjustment (still more to be made) with the folks that gross specimens however. Sometimes the tissues are still a bit too thick. Our pathologists come in between 7 and 8 am and leave mid-afternoon through early evening depending on the individual. We haven't made any changes in IHC or Special Stains methods. Oh, and for those that like to have their tissues tinted, the new Sakura process allows eosin to be added to the preprocessing solution. Hope this helps. Saundra ----- Original Message ----- From: "Jesus Ellin" To: Sent: Thursday, September 22, 2005 11:15 AM Subject: [Histonet] Microwave PROCCESSING > Nice to see that everyone made it back from NSH and are doing well. I > have a nice question that might start some serious discussion. Microwave > proccessing!!!! My question is how many people out there are doing it and > what issues did you have to work through to apply this technology to > everyday use??? How are you handling the testing of your immuno's since > you have to use tissue that has been run by the microwave proccessor??? > How is the transcription being done??? What time are the pathologist > coming in and how LATE are they staying?? Are you proccessing all tissue > type or only small biopies, breast biopsies, etc.?? What road block have > you come up against??? It this the rule to proccess all tissue or is this > the exception?? How are you handling reference lab consultation with your > tissue that is proccessed on the Microwave and sending it to a lab for > instance AFIP for consult and they only use the conventional proccessing?? > Is anyone out there doing both and how do they keep track of the different > types of tissue that have been proccessed within the different proccessor, > if you are indeed using both. What additional costs did you do you have, > with there new product from the proccessor?? > > > Our pathologist came back and are now foaming at the mouth for this > technology. We are not against this, but there are avenues that need to > be addressed, especially with IHC, FISH, and overall workflow. Any help > would greatly be appreciated and needed at this time. > > Your in Formalin > > Jesus Ellin > Yuma Regional Medical Center > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From histology <@t> 2hosts.com Mon Sep 26 20:31:03 2005 From: histology <@t> 2hosts.com (Histology) Date: Mon Sep 26 20:32:50 2005 Subject: [Histonet] Embedding centers References: <84BE46B37B314D409C5A17B7BAB022D622500A@IDC-EX-VS01.shriners.cc> Message-ID: <00a801c5c303$2329b020$c901fec0@FAMILYROOM> My favorite is still the Shandon, but we have both Shandon and Leica. I really don't like either the Leica or the Sakura models. Saundra Ellis Kaiser Permanente Histology Supervisor ----- Original Message ----- From: "Snider, Deanna" To: Sent: Thursday, September 22, 2005 11:54 AM Subject: [Histonet] Embedding centers > Hi everyone! I once again am in need of your knowledge and guidance! My > existing embedding center is on the fritz. We are looking into purchasing > a > new one. Which brands do you recommend? Why? I am a low volume research > lab, working with clinically engineered skin and human on mouse grafts. I > looked at the archives but not much was there! > Thanks in advance, > > Deanna Snider HT ASCP > Shriners Hospital for Children > Research Dept. > Cincinnati, Oh > 513-872-6388 > > CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may > contain confidential and privileged information for the use of the > designated recipients. If you are not the intended recipient, (or > authorized to receive for the recipient) you are hereby notified that you > have received this communication in error and that any review, disclosure, > dissemination, distribution or copying of it or its contents is > prohibited. > If you have received this communication in error, please destroy all > copies > of this communication and any attachments and contact the sender by reply > e-mail or telephone (813) 281-0300. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From samvaughnhisto <@t> gmail.com Mon Sep 26 21:59:41 2005 From: samvaughnhisto <@t> gmail.com (Sam Vaughn) Date: Mon Sep 26 21:59:53 2005 Subject: [Histonet] section edges lifting Message-ID: <1582e1c05092619596afc6ce1@mail.gmail.com> I'm new to histonet and have a question about histo staining. I have 6 um paraffin sections of mouse knee joints on uncoated superfrost plus slides. I collect the sections from a distilled water bath and let them drain at least 30min before placing them at 37 degrees overnight before staining. After Safranin O/ Fast Green staining a very thin area around the edge at the edge of the tissue lifts off the slide rendering it impossible to visualize with the microscope. Of course this is the the articular surface which we are interested in! I've used the same slides in the past without any coating and it has worked fine. I'm in a new lab now and trying to get things running. Any help you could give me would be really appreciated! Thanks, Sam From jkiernan <@t> uwo.ca Mon Sep 26 23:41:56 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Sep 26 23:42:21 2005 Subject: [Histonet] Pink precipitate is monoformazan from BCIP/NBT reaction onISH? References: Message-ID: <4338CD94.F2BDE464@uwo.ca> Dear Ralph, Your email is full of abbreviations and jargon. Am I right in thinking it's a question about localizing alkaline phosphatase activity by an indoxyl-tetrazolium method? If so, please provide a reference to the original publication and let us all know if you followed it exactly or made any changes. Part of your email suggests that the alkaline phosphatase activity is not endogenous but part of an amplification system used in in situ hybridization. The pH optima of endogenous and label alkaline phosphatases differ. The later paragraphs of your message indicate that you may not fully understand the significance of the mono- and diformazan products of reduction of nitro-BT. Both colours result from reduction of the tetrazolium salt, but you need controls to prove that the reduction was by bromochloroindoxyl and not by other reducing agents (such as diaphorases) in the tissue. Non-enzymatic reduction of tetrazolium salts by -SH has been a well known artifact {"nothing dehydrogenase") for 40-45 years. John A. Kiernan Anatomy Dept, UWO London, Canada. ___________________________________________________ Ralph Puchalski wrote: > > I am trying to figure out the origin of the pink precipitate in the > image I posted on http://www.histonet.org/site_images_frame.asp > > Please go to the image entitled: Pink Precipitate ver2.jpg. It is at > the top of the list on 9/26/05, posted by Ralph Puchalski. To see the > pink precipitate artifact, please open the image and set the scroll bars > on the bottom and right side of the image at their 1/2 way points. It > is ugly! > > I think this precipitate is the aggregation of the monoformazan > intermediate generated from NBT (after dephosphorylation of BCIP by > alkaline phosphatase) that is not completely reduced to diformazan, the > insoluble dark blue or black precipitate that labels cells expressing > target mRNAs in our in situ hybridization reactions. > > We don't know how the monoformazan adheres to the sections of tissue. > It appears to be non-covalent due to the tendency of the monoformazan to > migrate under the coverslip in the aqueous mounting medium that has yet > to dry, and form clumps or aggregates or pools as seen in the picture. > The monoformazan is soluble in ethanol so doesn't form pools or > aggregates. But as soon as it is exposed to an aqueous medium, it > precipitates. > > If we mount the post-ISH tissue sections with an organic based mounting > medium like UV-CureMount (Instrumedics), the monoformazan might cause > the entire section to turn to a brown tint as seen on > http://www.histonet.org/site_images_frame.asp > > Please go to the image Brown Tint 1.jpg. It is 4th image from the top > on 9/26, posted by Ralph Puchalski. The lower image is mounted with UV > CureMount, and the upper was with aqueous Hydro Matrix. There is no > pooling or precipitation of monoformazan with UV CureMount, but I think > it does cause the entire section to turn brown. > > Question: How do we eliminate this problem, which I believe is > monoformazan? If we reduce it fully using ascorbate, the section (later > mounted with HyrdoMatrix) turns dark blue or purple, the same color as > our ISH signal. We have tried washing off the monoformazan with 100% > ethanol prior to coverslipping, but only small amounts are removed. > 100% acetone also does not work effectively. > > Please let me know if you have any ideas that might help us eliminate > this problem. > > Thank you, > > Ralph > > Ralph Puchalski, Ph.D. > Manager, Process Engineering and Automation > Allen Institute for Brain Science > 551 N. 34th Street, Suite 200 > Seattle, WA 98103 > > ralphpu@alleninstitute.org > Tel: 206-548-7041 Fax: 206-548-7071 > www.brainatlas.org > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue Sep 27 02:46:21 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Tue Sep 27 02:49:01 2005 Subject: [Histonet] Silver ISH Message-ID: Does anyone out there have any info on silver ISH they could share? We hope to trial it for a research project on our Bench Mark XT and are waiting for Ventana to get back to us re- reagents but would appreciate it if anyone with any knowledge / experience of it would get back to me. Cheers Jacqui Malam Histopath Royal Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From arvind <@t> nbrc.res.in Tue Sep 27 03:22:05 2005 From: arvind <@t> nbrc.res.in (Arvind) Date: Tue Sep 27 03:19:51 2005 Subject: [Histonet] querry References: Message-ID: <001601c5c33c$8fc7d7a0$aa00a8c0@nbrc.res.in> can any one please tell me how to do antigen retrieval on soft tissue, what i do is i took cryosections on slide for human fetal brain tissue age upto 7 months (prenatal) sections are 40 micron thick, when i do HIER these sections are comming off the slides at some points, i use gelatin subbed slides for this please guide what to do in detail thanking in advance Arvind Singh Pundir National Brain Research Centre gurgaon, Haryana, INDIA arvind@nbrc.res.in From dellav <@t> musc.edu Tue Sep 27 07:10:27 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Tue Sep 27 07:10:07 2005 Subject: [Histonet] querry Message-ID: Antigen retrieval is undertaken to "un-do" the masking of tissue antigens that results from formalin (formaldehyde) fixation. fresh frozen tissues would generally not require this sort of pre-treatment.. unfixed tissues will not be resilient enough to withstand the high heat or harsh effects of extreme pH or proteolysis (depending upon the antigen retrieval method you use), hence you will see the sections come off the slides or look "chewed up" are you certain that retrieval pre-treatment is necessary in fresh frozen, unfixed tissues, for the antigens you are studying ? also, in general, thicker sections often have a greater tendency to lift off the slides. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Arvind" 09/27/05 04:23 AM >>> can any one please tell me how to do antigen retrieval on soft tissue, what i do is i took cryosections on slide for human fetal brain tissue age upto 7 months (prenatal) sections are 40 micron thick, when i do HIER these sections are comming off the slides at some points, i use gelatin subbed slides for this please guide what to do in detail thanking in advance Arvind Singh Pundir National Brain Research Centre gurgaon, Haryana, INDIA arvind@nbrc.res.in _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Tue Sep 27 08:10:12 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Sep 27 08:10:36 2005 Subject: [Histonet] querry In-Reply-To: Message-ID: <20050927131012.86957.qmail@web61219.mail.yahoo.com> I never did HIER of frozen sections. Neither on destained pap-smears we used for IHC tests. We always worked thin, air dried and hydrated FS. Rene J. Buesa Vinnie Della Speranza wrote: Antigen retrieval is undertaken to "un-do" the masking of tissue antigens that results from formalin (formaldehyde) fixation. fresh frozen tissues would generally not require this sort of pre-treatment.. unfixed tissues will not be resilient enough to withstand the high heat or harsh effects of extreme pH or proteolysis (depending upon the antigen retrieval method you use), hence you will see the sections come off the slides or look "chewed up" are you certain that retrieval pre-treatment is necessary in fresh frozen, unfixed tissues, for the antigens you are studying ? also, in general, thicker sections often have a greater tendency to lift off the slides. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Arvind" 09/27/05 04:23 AM >>> can any one please tell me how to do antigen retrieval on soft tissue, what i do is i took cryosections on slide for human fetal brain tissue age upto 7 months (prenatal) sections are 40 micron thick, when i do HIER these sections are comming off the slides at some points, i use gelatin subbed slides for this please guide what to do in detail thanking in advance Arvind Singh Pundir National Brain Research Centre gurgaon, Haryana, INDIA arvind@nbrc.res.in _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From rjbuesa <@t> yahoo.com Tue Sep 27 08:18:41 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Sep 27 08:19:07 2005 Subject: [Histonet] section edges lifting In-Reply-To: <1582e1c05092619596afc6ce1@mail.gmail.com> Message-ID: <20050927131841.19876.qmail@web61224.mail.yahoo.com> Check on the expirartion date of the slides. After some months they don't work as they are supposed to. Slides should be drained/dried on edge (not flat). A last resort would be to coverslip by hand, looking at the edges (and even flattening them out with a hair brush before coverslipping). Rene J. Buesa Sam Vaughn wrote: I'm new to histonet and have a question about histo staining. I have 6 um paraffin sections of mouse knee joints on uncoated superfrost plus slides. I collect the sections from a distilled water bath and let them drain at least 30min before placing them at 37 degrees overnight before staining. After Safranin O/ Fast Green staining a very thin area around the edge at the edge of the tissue lifts off the slide rendering it impossible to visualize with the microscope. Of course this is the the articular surface which we are interested in! I've used the same slides in the past without any coating and it has worked fine. I'm in a new lab now and trying to get things running. Any help you could give me would be really appreciated! Thanks, Sam _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From rjbuesa <@t> yahoo.com Tue Sep 27 08:21:54 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Sep 27 08:22:13 2005 Subject: [Histonet] IHC background problems on chicken embryos In-Reply-To: <000b01c5c300$225661f0$6a9a9618@Katri> Message-ID: <20050927132154.9220.qmail@web61217.mail.yahoo.com> I think that you should seek advise first from the DakoCytomation technical department. They are very helpful and for sure had experience with chicken (in Denmark). Rene J. Buesa Katri Tuomala wrote: Hi Histonetters, A colleague of mine is trying to get VWF (Dakocytomation) and VEGF(R&D Systems) working on chicken tissue. The antibodies are performing well on human and rat tissue, but excessive background is the problem with the chicken. Would the blocking serums (normal goat and normal rabbit respectively) be the problem as well as the biotinylated secondary reagents raised in goat and rabbit? My experience is strictly with human tissues, so I am stumped. To my knowledge neither of these antibodies have been proven to work with chicken, so maybe that is the problem. Any advice from "chicken" people? Katri Katri Tuomala Hamilton, Ontario, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From TJJ <@t> Stowers-Institute.org Tue Sep 27 08:37:06 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Sep 27 08:59:55 2005 Subject: [Histonet] Re: Pink precipitate... from BCIP/NBT reaction on ISH? Message-ID: Ralph, When we use BCIP/NBT, we do our color reaction development in the 37 degree C incubator in the dark. We change the solution frequently (every several hours, or when the solution turns pink). We do not lay the slides flat for their incubation, instead we use the 5-slide plastic mailers and have them standing upright. This keeps precipitate from settling on to the glass. Many of the chromogens for Alk. Phos. tend to form a precipitate, and the manufacturers don't recommend filtering. Your signal appears to be quite strong. Perhaps you can also decrease the time in chromogen? Best of luck to you, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From Dorothy.L.Webb <@t> HealthPartners.Com Tue Sep 27 09:03:32 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Tue Sep 27 09:04:04 2005 Subject: [Histonet] Iron stain Message-ID: <0E394B648E5284478A6CCB78E5AFDA2718C44E@hpes1.HealthPartners.int> I am having problems with the iron stain on bone marrow cores. We routinely run our irons on the Ventana Nexus, but, recently have had problems with the core biopsies picking up the "blue" staining in the histiocytes and other areas, making it hard for th pathologist to be confident in the diagnosis. Any suggestions as to why this is happening? I have tried running them down to sterile water and running the stain by hand using Mallory's Modification for iron and I am still seeing the staining being picked up in areas that should not be staining. HELP , and thanks!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From jnocito <@t> pathreflab.com Tue Sep 27 09:04:40 2005 From: jnocito <@t> pathreflab.com (Joe Nocito) Date: Tue Sep 27 09:05:32 2005 Subject: [Histonet] cpt codes for direct smears In-Reply-To: Message-ID: We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gsmith <@t> confocal.com Tue Sep 27 09:06:48 2005 From: gsmith <@t> confocal.com (Glenn Smith) Date: Tue Sep 27 09:07:32 2005 Subject: [Histonet] RE: Fluorescent Illumination Systems: Suggestions please? In-Reply-To: <200509270448.j8R4mL0F009876@mailstore2.execulink.net> Message-ID: <003301c5c36c$b83eb9d0$1f05a8c0@confocal.com> Melissa, If you haven't already, you may also want to post your question to a microscopy listserve too. For example, try http://listserv.acsu.buffalo.edu/cgi-bin/wa?A0=confocal. Glenn Smith, P.Eng. 519.886.9013 x38 (office) 519.498.0614 (mobile) Biomedical Photometrics Inc/GeneFocus Fluorescence and Brightfield Laser Scanning Instruments and Software for Pathology -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, September 27, 2005 12:48 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 22, Issue 33 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: antigen retrieval for IHC (May Wei) 2. RE: antigen retrieval for IHC (Morken, Tim - Labvision) 3. Fluorescent Illumination Systems: Suggestions please? (Melissa Gonzalez) 4. Re: cryostat decontamination (Instrumedics) 5. RE: antigen retrieval for IHC (Pamela Marcum) 6. Low temp Antigen Retrieval for Bone Sections (Kim Merriam) 7. Re: Fluorescent Illumination Systems: Suggestions please? (kgibbon@qltinc.com) 8. RE: Low temp Antigen Retrieval for Bone Sections (Monfils, Paul) 9. cpt codes for direct smears (Barb Richmond) 10. Re: antigen retrieval for IHC (Bryan Hewlett) 11. slides and RE: [Histonet] Low temp Antigen Retrieval for Bone Sections (Gayle Callis) 12. RE: slide QC (Baez, Janet) 13. Cryostat decontamination (Jan.Minshew@leica-microsystems.com) 14. RE: Posting for Histology List Serve (Nelson, Sandy - PPH) 15. Staining fibrin - thrombus (Andrea T. Hooper) 16. IHC background problems on chicken embryos (Katri Tuomala) 17. Pink precipitate is monoformazan from BCIP/NBT reaction on ISH? (Ralph Puchalski) 18. Re: Microwave PROCCESSING (Histology) 19. Re: Embedding centers (Histology) 20. section edges lifting (Sam Vaughn) 21. Re: Pink precipitate is monoformazan from BCIP/NBT reaction onISH? (John Kiernan) ---------------------------------------------------------------------- Message: 1 Date: Mon, 26 Sep 2005 10:05:15 -0700 From: "May Wei" Subject: RE: [Histonet] antigen retrieval for IHC To: Cc: histonet@lists.utsouthwestern.edu Message-ID: <37DC9F93CF7F864182D0463EF93D571B0B5043@ISLETON2.california.biogenex.com> Content-Type: text/plain; charset="us-ascii" One of papers with Shi/Taylor is "Standardization of Immunohistochemistry Based on Antigen Retrieval Technique for Routine Formalin-fixed Tissue Sections" in Applied Immunohistochemistry 6(2): 89-96, 1998. I have the article. I also have the list of AR paper references. If you like to receive it, I can send to you. May Wei -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Sunday, September 25, 2005 11:14 PM To: Maria Mejia Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] antigen retrieval for IHC Maria, can you give chapter and verse for the Shi/Taylor and Puchrler/Meloan papers? Most papers with Shi and Taylor among the authors are about high temperature antigen retrieval (boiling water, with pH optima for various antigens). Holde Puchtler and Susan Meloan published many imortant papers about staining ad histochemistry in the 1970s=1980s. Susan M was a histonetter in the 1990s. ***************** Maria Mejia wrote: > > Hello, > > I believe it was sometime in June of this year that there was a > discussion on the histonet regarding "the simplest form" of reversing > the effects for formalin fixation on tissue was to wash the tissue > well (before) processing. > > Well, I just read the article "Antigen retrieval IHC: Past, Present, & > Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google > this article). It's a very interesting! Anyway, the article has a > section titled non-heating AR method stating (this) same simple method > by (Puchtler & Meloan). It also goes on to say that Elias JM (1990) > adopted this method routinely by storing deparaffinized in fresh > changes of 10% sucrose/PBS @ 4C overnight (before) IHC. > > My questions are, does anyone know or tried this method used by Elias? > And, can anyone explain the mechanism of 10% sucrose/PBS play in this > AR method? > > Just curious! > > Maria Bartola Mejia > Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 > Email: maria@ski.org > Phone: (415)-345-2185 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Mon, 26 Sep 2005 13:12:32 -0400 From: "Morken, Tim - Labvision" Subject: RE: [Histonet] antigen retrieval for IHC To: histonet@lists.utsouthwestern.edu Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D178@usca0082k08.labvision.apogent.com> Content-Type: text/plain It's true that most papers by Shi et al are on high-temperature antigen retrieval, but in their book on antigen retrieval (Antigen retrieval Techniques, Immunohistochemistry and Molecular Morphology, ed. Shan-Rong Shi et al, Eaton Publishing, 2000) they make clear that the term "antigen retrieval" should cover all types -enzyme, heat, or other. In fact they make a plea for everyone publishing methods to use the term "antigen retrieval" so matter what the method so that databases are easy to search. With the plethora of terms used to decribe the methods it is nearly impossible to find all the methods developed in the past 14 years. Part of the reason for people using all the different terms was the assumption by many that BioGenex had trademarked the term. BioGenex did apply for a trademark, but then withdrew it. So the term is open to all to use. However, BioGenex does have a patent on a very specific method of antigen retrieval, and some antigen retrieval buffer formulations. Tim Morken Lab Vision - Neomarkers www.labvision.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Sunday, September 25, 2005 11:14 PM To: Maria Mejia Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] antigen retrieval for IHC Maria, can you give chapter and verse for the Shi/Taylor and Puchrler/Meloan papers? Most papers with Shi and Taylor among the authors are about high temperature antigen retrieval (boiling water, with pH optima for various antigens). Holde Puchtler and Susan Meloan published many imortant papers about staining ad histochemistry in the 1970s=1980s. Susan M was a histonetter in the 1990s. ***************** Maria Mejia wrote: > > Hello, > > I believe it was sometime in June of this year that there was a > discussion on the histonet regarding "the simplest form" of reversing > the effects for formalin fixation on tissue was to wash the tissue > well (before) processing. > > Well, I just read the article "Antigen retrieval IHC: Past, Present, & > Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can google > this article). It's > a very interesting! Anyway, the article has a section titled non-heating > AR method stating (this) same simple method by (Puchtler & Meloan). It also > goes on to say that Elias JM (1990) adopted this method routinely by storing > deparaffinized in fresh changes of 10% sucrose/PBS @ 4C overnight > (before) IHC. > > My questions are, does anyone know or tried this method used by Elias? > And, can anyone explain the mechanism of 10% sucrose/PBS play in this > AR method? > > Just curious! > > Maria Bartola Mejia > Smith-Kettlewell Eye Research Institute > San Francisco, CA 94115 > Email: maria@ski.org > Phone: (415)-345-2185 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 26 Sep 2005 10:50:34 -0700 From: "Melissa Gonzalez" Subject: [Histonet] Fluorescent Illumination Systems: Suggestions please? To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello all, Sorry to re-post, but I only received one response to the following: I am currently in the market to upgrade our existing HB0 100 lamp house, which is very old, to several of new systems. Either the newest version of the HB0 100 from Zeiss, a mercury halide system called X-Cite from EXFO, or a xenon based burner called Lambda LS from Sutter Instruments. They are all comparable in price, but my main concern is increasing the brightness of my signal and evenly illuminating the field of view. I mostly look at DAPI, eGFP, FITC, and Alexa 594, which from spectral outputs, each one of these systems appears to have a different weakness in one of the above. On a plus, the Xcite rates their bulbs at 1500 hours, and the Lambda at 500-1000hrs. Does anyone out there have any input/comments/experience with these systems? (This would be hooked up to a Zeiss Axioplan) Please let me know if you have any info, as I have been unsuccessful so far with demos and need to place an order soon. Thanks a lot -I really appreciate it, Melissa ------------------------------ Message: 4 Date: Mon, 26 Sep 2005 13:53:16 -0400 From: "Instrumedics" Subject: Re: [Histonet] cryostat decontamination To: Cc: HistoNet Server Message-ID: <01d401c5c2c3$336ba0f0$6401a8c0@INSTRUMEDICS22> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Instrumedics' Cryo-Vac-Away keeps the cryostat free of trimming debris. The debris is suctioned away as it is generated at the block face when you trim the block. The debris is captured in a primary filter that is inside the cryostat and downstream of the primary filter is a viral/ bacterial filter that captures pathogens. The cryostat remains spotless and substantially reduces the hazards associated with frozen sectioning. Please visit www.instrumedics.com for details. Bernice ----- Original Message ----- From: To: Sent: Monday, September 26, 2005 10:59 AM Subject: Re: [Histonet] cryostat decontamination > Greetings, > I recently attended Gloria Limetti's seminar at NSH about cryostat and > microtome features. She is from the University of Pitssburgh. Attendees > were > given an extensive spreadsheet listing which features were available > on > which > models. In an effort not to come across as non vendor specific, Gloria > assigned all the companies letters. Just looking over the features, it's > fairly > easy to decipher which company is which. I don't have my copy in front > of me > now but I believe there are 7 models with various types of > decontamination > systems offered. > Hacker Instruments SL5000 is one of the units that is available with an > automatic decontamination feature, perhaps Gloria will post the names of > the > other companies. > As for UV decontamination, it is my understanding the the UV light is only > effective on the surfaces of the cryostat and microtome chamber where the > light > actually comes into contact. Nooks and cranies would still need to be > wiped down manually. As with Vinnie, I would be interested in reading a > study or > two on it's effectiveness in histology applications. > Regards, > > Dorothy Murphy Traczyk > National Sales Manager > Hacker Instruments & Industries Inc. > PO Box 1176 > Winnsboro, SC 29180 > 1-800-442-2537 > hackerlab@aol.com > _www.hacker_ (http://www.hacker/) insruments.com > > ------------------------------ Message: 5 Date: Mon, 26 Sep 2005 14:01:50 -0400 From: Pamela Marcum Subject: RE: [Histonet] antigen retrieval for IHC To: "Morken, Tim - Labvision" , histonet@lists.utsouthwestern.edu Message-ID: <6.1.1.1.2.20050926140018.01a53ce0@mail.vet.upenn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Thanks for the update Tim. I know for years we have all been careful about using antigen retrieval due to the BioGenex issue. They do have a patent on the one tyoe and it was spread to all for a while. Pam Marcum At 01:12 PM 9/26/2005, Morken, Tim - Labvision wrote: >It's true that most papers by Shi et al are on high-temperature antigen >retrieval, but in their book on antigen retrieval (Antigen retrieval >Techniques, Immunohistochemistry and Molecular Morphology, ed. >Shan-Rong Shi et al, Eaton Publishing, 2000) they make clear that the >term "antigen retrieval" should cover all types -enzyme, heat, or >other. In fact they make a plea for everyone publishing methods to use >the term "antigen retrieval" so matter what the method so that >databases are easy to search. With the plethora of terms used to >decribe the methods it is nearly impossible to find all the methods >developed in the past 14 years. Part of the reason for people using all >the different terms was the assumption by many that BioGenex had >trademarked the term. BioGenex did apply for a trademark, but then >withdrew it. So the term is open to all to use. However, BioGenex does >have a patent on a very specific method of antigen retrieval, and some >antigen retrieval buffer formulations. > >Tim Morken >Lab Vision - Neomarkers >www.labvision.com > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John >Kiernan >Sent: Sunday, September 25, 2005 11:14 PM >To: Maria Mejia >Cc: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] antigen retrieval for IHC > > >Maria, can you give chapter and verse for the Shi/Taylor >and Puchrler/Meloan papers? Most papers with Shi and Taylor among the >authors are about high temperature antigen retrieval (boiling water, >with pH optima for various antigens). > >Holde Puchtler and Susan Meloan published many imortant papers about >staining ad histochemistry in the 1970s=1980s. Susan M was a >histonetter in the 1990s. > >***************** > >Maria Mejia wrote: > > > > Hello, > > > > I believe it was sometime in June of this year that there was a > > discussion on the histonet regarding "the simplest form" of > > reversing the effects for formalin fixation on tissue was to wash > > the tissue well (before) processing. > > > > Well, I just read the article "Antigen retrieval IHC: Past, Present, > > & Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can > > google this article). It's a very interesting! Anyway, the article > > has a section titled non-heating AR method stating (this) same > > simple method by (Puchtler & Meloan). It >also > > goes on to say that Elias JM (1990) adopted this method routinely by >storing > > deparaffinized in fresh changes of 10% sucrose/PBS @ 4C overnight > > (before) IHC. > > > > My questions are, does anyone know or tried this method used by > > Elias? And, can anyone explain the mechanism of 10% sucrose/PBS play > > in this AR method? > > > > Just curious! > > > > Maria Bartola Mejia > > Smith-Kettlewell Eye Research Institute > > San Francisco, CA 94115 > > Email: maria@ski.org > > Phone: (415)-345-2185 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu ------------------------------ Message: 6 Date: Mon, 26 Sep 2005 11:34:06 -0700 (PDT) From: Kim Merriam Subject: [Histonet] Low temp Antigen Retrieval for Bone Sections To: Histonet Message-ID: <20050926183406.77615.qmail@web50312.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello All, I will be performing some IHC on mouse femurs (about 12 or so different antibodies). In the past, I have had a lot of trouble keeping these sections on the slides during HIER. I did a quick search on the archives and there were lots of suggestions, but nothing definitive. I was reading an article about "low temp AR", overnight at 60C in TEG buffer, pH 9.0. Has anyone tried this method? Also - what does TEG buffer stand for? Any help would be greatly appreciated. Kim Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 7 Date: Mon, 26 Sep 2005 12:22:30 -0700 From: kgibbon@qltinc.com Subject: Re: [Histonet] Fluorescent Illumination Systems: Suggestions please? To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi Melissa, I have used the Sutter xenon Lambda unit for a few years now and really like it, The spectrum is much more even than a standard mercury bulb. I have the model with the extended bandwidth so I could use it well into the UV wavelengths but it does produce ozone which must be vented, I believe they have another model that does not do this but has a higher minimum wavelength. Your choice, depending what fluorochromes you want to light up. Kevin Gibbon QLT Inc. |---------+-------------------------------------------> | | "Melissa Gonzalez" | | | | | | Sent by: | | | | | | | | | | | | 09/26/2005 10:50 AM | | | | |---------+-------------------------------------------> >--------------------------------------------------------------------------- ---------------------------------------------------| | | | To: | | cc: | | Subject: [Histonet] Fluorescent Illumination Systems: Suggestions please? | >--------------------------------------------------------------------------- ---------------------------------------------------| Hello all, Sorry to re-post, but I only received one response to the following: I am currently in the market to upgrade our existing HB0 100 lamp house, which is very old, to several of new systems. Either the newest version of the HB0 100 from Zeiss, a mercury halide system called X-Cite from EXFO, or a xenon based burner called Lambda LS from Sutter Instruments. They are all comparable in price, but my main concern is increasing the brightness of my signal and evenly illuminating the field of view. I mostly look at DAPI, eGFP, FITC, and Alexa 594, which from spectral outputs, each one of these systems appears to have a different weakness in one of the above. On a plus, the Xcite rates their bulbs at 1500 hours, and the Lambda at 500-1000hrs. Does anyone out there have any input/comments/experience with these systems? (This would be hooked up to a Zeiss Axioplan) Please let me know if you have any info, as I have been unsuccessful so far with demos and need to place an order soon. Thanks a lot -I really appreciate it, Melissa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This electronic transmission (including any and all attachments) is intended solely for the use of the individual or entity to whom it is addressed and may contain information that is privileged and/or confidential. If you are not the intended recipient of this electronic transmission, you are hereby notified that any disclosure, copying or distribution, or the taking of any action in reliance upon the contents of this electronic transmission, is strictly prohibited, and you are further requested to purge this electronic transmission and all copies thereof from your computer system. ------------------------------ Message: 8 Date: Mon, 26 Sep 2005 15:59:28 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] Low temp Antigen Retrieval for Bone Sections To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175E3@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" I haven't tried this method of antigen retrieval, but TEG buffer is Tris-EGTA buffer. The usual formula (I have seen a couple of variations) is: Distilled water 1,000 ml Trisma base 1.21 gram EGTA 0.19 gram The usual target pH is between 8.95 and 9.0 EGTA is ethylene glycol bis(B-aminoethyl)-N,N,N',N' tetraacetic acid. (The capital "B" in the above name represents "beta". My email program doesn't support the symbol.) > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kim > Merriam > Sent: Monday, September 26, 2005 11:34 AM > To: Histonet > Subject: [Histonet] Low temp Antigen Retrieval for Bone Sections > > Hello All, > > I will be performing some IHC on mouse femurs (about 12 or so > different antibodies). In the past, I have had a lot of trouble > keeping these sections on the slides during HIER. I did a quick > search on the archives and there were lots of suggestions, but nothing > definitive. > > I was reading an article about "low temp AR", overnight at 60C in TEG > buffer, pH 9.0. Has anyone tried this method? Also - what does TEG > buffer stand for? > > Any help would be greatly appreciated. > > Kim > > > Kim Merriam > Novartis > Cambridge, MA __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 9 Date: Mon, 26 Sep 2005 15:29:00 -0500 From: "Barb Richmond" Subject: [Histonet] cpt codes for direct smears To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? ------------------------------ Message: 10 Date: Mon, 26 Sep 2005 16:31:28 -0400 From: "Bryan Hewlett" Subject: Re: [Histonet] antigen retrieval for IHC To: "Patsy Ruegg" , , "'Maria Mejia'" Cc: histonet@lists.utsouthwestern.edu Message-ID: <002101c5c2d9$467fce80$6400a8c0@mainbox> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Patsy, You will need to wash MUCH longer than 2 hours in running tap water in order to obtain significant reversal after I year in NBF. The Helander paper quoted in my NSH handout gives about 6 days to effect a 90% reversal and that was following 24 hour fixation. Best regards, Bryan ----- Original Message ----- From: "Patsy Ruegg" To: ; "'Maria Mejia'" Cc: Sent: Monday, September 26, 2005 11:58 AM Subject: RE: [Histonet] antigen retrieval for IHC > Shi mentions in his book on pg.10 ANTIGEN RESTORATION > "Simply bathing deparaffinized sections in a cold 20% sucrose-saline > solution could, over several days, restore a certain amount of > immunoreactivity." > > In Introduction to IHC by Polak and Van Noorden on pg 24 3.8.1 "The > simplest > form of reversing the effects of formalin is to wash the tissue well > before > processing" > > I am about to test these statements, as I just received tissues for an > IHC project that have been in 10% NBF for over one year. I washed the > tissues in running tap h20 for 2 hrs., I will now process them into > paraffin. I am planning as well to put some of the sections in 20% > sucrose at 4dc for 2 days if I have trouble getting good IHC signal. > I will keep you all posted > on this experiment. These are samples of mouse mammory tissue. I will be > testing them with cleaved caspase 3 and Ki67 to start. I have both of > these > antibodies worked out very well for ffpe tissues fixed 24-48 hrs. > > Patsy > > > Patsy Ruegg, HT(ASCP)QIHC > IHCtech, LLC > Fitzsimmons BioScience Park > 12635 Montview Blvd. Suite 216 > Aurora, CO 80010 > P-720-859-4060 > F-720-859-4110 > wk email pruegg@ihctech.net > web site www.ihctech.net > > > This email is confidential and intended solely for the use of the > Person(s) > ('the intended recipient') to whom it was addressed. Any views or opinions > presented are solely those of the author. It may contain information that > is > privileged & confidential within the meaning of applicable law. > Accordingly > any dissemination, distribution, copying, or other use of this message, or > any of its contents, by any person other than the intended recipient may > constitute a breach of civil or criminal law and is strictly prohibited. > If > you are NOT the intended recipient please contact the sender and dispose > of > this e-mail as soon as possible. > > > -----Original Message----- > From: John Kiernan [mailto:jkiernan@uwo.ca] > Sent: Sunday, September 25, 2005 11:14 PM > To: Maria Mejia > Cc: pruegg@ihctech.net; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] antigen retrieval for IHC > > Maria, can you give chapter and verse for the Shi/Taylor and > Puchrler/Meloan > papers? Most papers with Shi and Taylor among the authors are about high > temperature antigen retrieval (boiling water, with pH optima for various > antigens). > > Holde Puchtler and Susan Meloan published many imortant papers about > staining ad histochemistry in the 1970s=1980s. Susan M was a > histonetter in the 1990s. > > ***************** > > Maria Mejia wrote: >> >> Hello, >> >> I believe it was sometime in June of this year that there was a >> discussion on the histonet regarding "the simplest form" of reversing >> the effects for formalin fixation on tissue was to wash the tissue >> well (before) processing. >> >> Well, I just read the article "Antigen retrieval IHC: Past, Present, >> & Future by Shan-Rong Shi, richard J. Cote & Clive R. Taylor (can >> google this article). It's a very interesting! Anyway, the article >> has a section titled non-heating AR method stating (this) same simple >> method by (Puchtler & Meloan). It also goes on to say that Elias JM >> (1990) adopted this method routinely by storing deparaffinized in >> fresh changes of 10% sucrose/PBS @ 4C overnight (before) IHC. >> >> My questions are, does anyone know or tried this method used by >> Elias? And, can anyone explain the mechanism of 10% sucrose/PBS play >> in this AR method? >> >> Just curious! >> >> Maria Bartola Mejia >> Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 >> Email: maria@ski.org >> Phone: (415)-345-2185 >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 11 Date: Mon, 26 Sep 2005 15:04:48 -0600 From: Gayle Callis Subject: slides and RE: [Histonet] Low temp Antigen Retrieval for Bone Sections To: "Monfils, Paul" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050926150104.01b40890@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I strongly suggest if you try this retrieval, that you use Erie Scientific EXCEL slides, which are coated specially to reduce the effects of chelator based retrieval solutions at high aka alkaline pH. Many people experience section loss with this pH and chelator, so Erie developed a new positively charged coating to counteract losing precious sections. You can access samples from them to see if they work out for you, contact your Erie sales rep or even Erie directly, they have a website. Biogenex also has a decalcified bone retrieval solution, another one to try out. At 01:59 PM 9/26/2005, you wrote: >I haven't tried this method of antigen retrieval, but TEG buffer is >Tris-EGTA buffer. The usual formula (I have seen a couple of >variations) is: > >Distilled water 1,000 ml >Trisma base 1.21 gram >EGTA 0.19 gram > >The usual target pH is between 8.95 and 9.0 > >EGTA is ethylene glycol bis(B-aminoethyl)-N,N,N',N' tetraacetic acid. > >(The capital "B" in the above name represents "beta". My email program >doesn't support the symbol.) > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kim > > Merriam > > Sent: Monday, September 26, 2005 11:34 AM > > To: Histonet > > Subject: [Histonet] Low temp Antigen Retrieval for Bone Sections > > > > Hello All, > > > > I will be performing some IHC on mouse femurs (about 12 or so > > different antibodies). In the past, I have had a lot of trouble > > keeping these sections on the slides during HIER. I did a quick > > search on the archives and there were lots of suggestions, but > > nothing definitive. > > > > I was reading an article about "low temp AR", overnight at 60C in > > TEG buffer, pH 9.0. Has anyone tried this method? Also - what does > > TEG buffer stand for? > > > > Any help would be greatly appreciated. > > > > Kim > > > > > > Kim Merriam > > Novartis > > Cambridge, MA __________________________________________________ > > Do You Yahoo!? > > Tired of spam? Yahoo! Mail has the best spam protection around > > http://mail.yahoo.com > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 12 Date: Mon, 26 Sep 2005 14:08:53 -0700 From: "Baez, Janet" Subject: RE: [Histonet] slide QC To: "Bernadette Weston" , Message-ID: <9E956D8FEB06C2408B08AC16498325E90139FA@adsl-67-113-77-28.dsl.lsan03.pacbell .net> Content-Type: text/plain; charset="us-ascii" We run one slide of an appendix everyday and date the slide. The histologist evaluates the slide before running the rest of the workload. The Pathologists then documents the quality on our QC sheet. -----Original Message----- From: Bernadette Weston [mailto:bernaweston@hotmail.com] Sent: Monday, September 26, 2005 4:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] slide QC What method of QC checking slides(H&E) do you use, does the histologist check the slides microscopically before they take them to the pathologist or does the pathologist review them and give either a verbal or written evaluation of the day's work? If the histologist does the before check, how many slides are reviewed? Bernadette Weston HT Histology Supervisor Barberton Citizens Hospital Barberton,OH _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Mon, 26 Sep 2005 17:14:38 -0500 From: Jan.Minshew@leica-microsystems.com Subject: [Histonet] Cryostat decontamination To: Traczyk7@aol.com Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi Dorothy and Histonet subscribers, I'm writing in response to comments I've seen in the past few days about UV disinfection procedures for cryostats. I'm not sure if the listing on the spreadsheet from Gloria's workshop referred to our instrument or not, but Leica recently released the CM1850 UV cryostat with ultraviolet light (UVC) disinfection...so I thought I should chime in. We had the same questions about cryostat disinfection that many of you have had. Therefore, as part of our development process we hired an independent laboratory to perform tests that would verify the efficacy of UVC exposure against specific pathogens in the CM1850 at cold temperatures. The results assured us that our system is a convenient and safe means of inactivating microorganisms in the air and on exposed surfaces at temperatures down to -20?C and that using the system significantly reduces the risk of infection to the operator. We proudly provide a CD that contains the certificate from the consultant, details of how the tests were performed and the results that were obtained. Whether a cryostat has a built-in disinfection system of any kind or not, there are several very important things to remember about disinfecting cryostats. 1. Before beginning a disinfection protocol, don personal protective equipment (puncture and penetration resistant gloves, gowns, etc). 2. Remove all debris from the cryostat and disposed of it according to the policies and procedures of your institution. The debris must be removed because organic material (blood and proteins) may contain high concentrations of microorganisms and could possibly inactivate chemical germicides or prevent access to contaminated surfaces. 3. Use 70% ethyl alcohol to clean the cryostat. The germicidal activity of ethyl alcohol is most effective in that range and it has an advantage over isopropyl alcohol of being able to kill hydrophilic viruses. 4. For those of us in the USA (other countries have access to other products), the EPA maintains a list of Antimicrobial Chemical/Registration Number Indexes and it is posted on their website http://www.epa.gov/oppad001/chemregindex.htm. From this link you can find agents effective against bloodborne pathogens such as Mycobacterium tuberculosis, human HIV-1 virus, Hepatitis B or Hepatitis C virus. It is critical to remember that NONE of these solutions have been tested at low temperatures and they can only be used at room temperature. 5. Do not create aerosols by spraying disinfectant (or anything else) in an open cryostat chamber. Pour disinfectants onto absorbent disposable towels and allow them to remain in contact with contaminated surfaces for the length of time specified in the instructions of the individual agents. I hope this information is useful. Please let me know if you have any questions. Best wishes to all, Jan Minshew HT, HTL(ASCP) Marketing Manager Leica Microsystems, Inc. 2345 Waukegan Rd Bannockburn, IL 60015 800.248.0123 x7051 Traczyk7@aol.com Sent by: To: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: (bcc: Jan Minshew/USDER/West/Leica) western.edu Subject: Re: [Histonet] cryostat decontamination 09/26/2005 09:59 AM Greetings, I recently attended Gloria Limetti's seminar at NSH about cryostat and microtome features. She is from the University of Pitssburgh. Attendees were given an extensive spreadsheet listing which features were available on which models. In an effort not to come across as non vendor specific, Gloria assigned all the companies letters. Just looking over the features, it's fairly easy to decipher which company is which. I don't have my copy in front of me now but I believe there are 7 models with various types of decontamination systems offered. Hacker Instruments SL5000 is one of the units that is available with an automatic decontamination feature, perhaps Gloria will post the names of the other companies. As for UV decontamination, it is my understanding the the UV light is only effective on the surfaces of the cryostat and microtome chamber where the light actually comes into contact. Nooks and cranies would still need to be wiped down manually. As with Vinnie, I would be interested in reading a study or two on it's effectiveness in histology applications. Regards, Dorothy Murphy Traczyk National Sales Manager Hacker Instruments & Industries Inc. PO Box 1176 Winnsboro, SC 29180 1-800-442-2537 hackerlab@aol.com _www.hacker_ (http://www.hacker/) insruments.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com ------------------------------ Message: 14 Date: Mon, 26 Sep 2005 18:03:15 -0500 From: "Nelson, Sandy - PPH" Subject: [Histonet] RE: Posting for Histology List Serve To: "Histology List Serve" Message-ID: Content-Type: text/plain; charset=iso-8859-1 > Subject: Histology Management Position > > A 500 bed hospital adjacent to the Houston Medical Center has an opening for a full time Histology Supervisor. This candidate should have strong leadership skills and prior management experience is preferred. Interested? Call 713.527.5292. > > Sandra Nelson > Director Laboratory Services > Park Plaza Hospital > Houston, Texas > 713.527.5292 > sandy.nelson@tenethealth.com > ** This is not intended to be a legally binding or legally effective signature > > > ------------------------------ Message: 15 Date: Mon, 26 Sep 2005 20:27:30 -0400 From: "Andrea T. Hooper" Subject: [Histonet] Staining fibrin - thrombus To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Hi All! If you wanted to stain a paraffin section (frozen also available) of mouse tissue and prove what you are seeing is a result of a thrombotic event what stains would you use? I was thinking to do an anti-fibrin immunostain (antibody suggestions welcome) and perhaps some sort of fibrin special stain (I came up with Carstairs' and Weigert's as probably being the best). Any recommendations for antibodies, techniques, protocols, words of wisdom? Please any information, detailed or not, will be put to good use ... and thanks! Andrea -- ------------------------------ Message: 16 Date: Mon, 26 Sep 2005 21:09:38 -0400 From: "Katri Tuomala" Subject: [Histonet] IHC background problems on chicken embryos To: "HistoNet Server" Message-ID: <000b01c5c300$225661f0$6a9a9618@Katri> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Hi Histonetters, A colleague of mine is trying to get VWF (Dakocytomation) and VEGF(R&D Systems) working on chicken tissue. The antibodies are performing well on human and rat tissue, but excessive background is the problem with the chicken. Would the blocking serums (normal goat and normal rabbit respectively) be the problem as well as the biotinylated secondary reagents raised in goat and rabbit? My experience is strictly with human tissues, so I am stumped. To my knowledge neither of these antibodies have been proven to work with chicken, so maybe that is the problem. Any advice from "chicken" people? Katri Katri Tuomala Hamilton, Ontario, Canada ------------------------------ Message: 17 Date: Mon, 26 Sep 2005 18:29:53 -0700 From: "Ralph Puchalski" Subject: [Histonet] Pink precipitate is monoformazan from BCIP/NBT reaction on ISH? To: Message-ID: Content-Type: text/plain; charset="us-ascii" I am trying to figure out the origin of the pink precipitate in the image I posted on http://www.histonet.org/site_images_frame.asp Please go to the image entitled: Pink Precipitate ver2.jpg. It is at the top of the list on 9/26/05, posted by Ralph Puchalski. To see the pink precipitate artifact, please open the image and set the scroll bars on the bottom and right side of the image at their 1/2 way points. It is ugly! I think this precipitate is the aggregation of the monoformazan intermediate generated from NBT (after dephosphorylation of BCIP by alkaline phosphatase) that is not completely reduced to diformazan, the insoluble dark blue or black precipitate that labels cells expressing target mRNAs in our in situ hybridization reactions. We don't know how the monoformazan adheres to the sections of tissue. It appears to be non-covalent due to the tendency of the monoformazan to migrate under the coverslip in the aqueous mounting medium that has yet to dry, and form clumps or aggregates or pools as seen in the picture. The monoformazan is soluble in ethanol so doesn't form pools or aggregates. But as soon as it is exposed to an aqueous medium, it precipitates. If we mount the post-ISH tissue sections with an organic based mounting medium like UV-CureMount (Instrumedics), the monoformazan might cause the entire section to turn to a brown tint as seen on http://www.histonet.org/site_images_frame.asp Please go to the image Brown Tint 1.jpg. It is 4th image from the top on 9/26, posted by Ralph Puchalski. The lower image is mounted with UV CureMount, and the upper was with aqueous Hydro Matrix. There is no pooling or precipitation of monoformazan with UV CureMount, but I think it does cause the entire section to turn brown. Question: How do we eliminate this problem, which I believe is monoformazan? If we reduce it fully using ascorbate, the section (later mounted with HyrdoMatrix) turns dark blue or purple, the same color as our ISH signal. We have tried washing off the monoformazan with 100% ethanol prior to coverslipping, but only small amounts are removed. 100% acetone also does not work effectively. Please let me know if you have any ideas that might help us eliminate this problem. Thank you, Ralph Ralph Puchalski, Ph.D. Manager, Process Engineering and Automation Allen Institute for Brain Science 551 N. 34th Street, Suite 200 Seattle, WA 98103 ralphpu@alleninstitute.org Tel: 206-548-7041 Fax: 206-548-7071 www.brainatlas.org ------------------------------ Message: 18 Date: Mon, 26 Sep 2005 18:28:56 -0700 From: "Histology" Subject: Re: [Histonet] Microwave PROCCESSING To: Message-ID: <009f01c5c302$d761a9e0$c901fec0@FAMILYROOM> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original We are using microwave processing. We've had to process fatty tissues, lymph nodes and cell blocks on a standard VIP, but are using the Sakura Xpress for the rest. We've recently had a software and reagent update which will allow all tissues types to run on the standard 1.5 hour program. There was some adjustment (still more to be made) with the folks that gross specimens however. Sometimes the tissues are still a bit too thick. Our pathologists come in between 7 and 8 am and leave mid-afternoon through early evening depending on the individual. We haven't made any changes in IHC or Special Stains methods. Oh, and for those that like to have their tissues tinted, the new Sakura process allows eosin to be added to the preprocessing solution. Hope this helps. Saundra ----- Original Message ----- From: "Jesus Ellin" To: Sent: Thursday, September 22, 2005 11:15 AM Subject: [Histonet] Microwave PROCCESSING > Nice to see that everyone made it back from NSH and are doing well. I > have a nice question that might start some serious discussion. Microwave > proccessing!!!! My question is how many people out there are doing it and > what issues did you have to work through to apply this technology to > everyday use??? How are you handling the testing of your immuno's since > you have to use tissue that has been run by the microwave proccessor??? > How is the transcription being done??? What time are the pathologist > coming in and how LATE are they staying?? Are you proccessing all tissue > type or only small biopies, breast biopsies, etc.?? What road block have > you come up against??? It this the rule to proccess all tissue or is this > the exception?? How are you handling reference lab consultation with your > tissue that is proccessed on the Microwave and sending it to a lab for > instance AFIP for consult and they only use the conventional proccessing?? > Is anyone out there doing both and how do they keep track of the different > types of tissue that have been proccessed within the different proccessor, > if you are indeed using both. What additional costs did you do you have, > with there new product from the proccessor?? > > > Our pathologist came back and are now foaming at the mouth for this > technology. We are not against this, but there are avenues that need to > be addressed, especially with IHC, FISH, and overall workflow. Any help > would greatly be appreciated and needed at this time. > > Your in Formalin > > Jesus Ellin > Yuma Regional Medical Center > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 19 Date: Mon, 26 Sep 2005 18:31:03 -0700 From: "Histology" Subject: Re: [Histonet] Embedding centers To: Message-ID: <00a801c5c303$2329b020$c901fec0@FAMILYROOM> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original My favorite is still the Shandon, but we have both Shandon and Leica. I really don't like either the Leica or the Sakura models. Saundra Ellis Kaiser Permanente Histology Supervisor ----- Original Message ----- From: "Snider, Deanna" To: Sent: Thursday, September 22, 2005 11:54 AM Subject: [Histonet] Embedding centers > Hi everyone! I once again am in need of your knowledge and guidance! My > existing embedding center is on the fritz. We are looking into purchasing > a > new one. Which brands do you recommend? Why? I am a low volume research > lab, working with clinically engineered skin and human on mouse grafts. I > looked at the archives but not much was there! > Thanks in advance, > > Deanna Snider HT ASCP > Shriners Hospital for Children > Research Dept. > Cincinnati, Oh > 513-872-6388 > > CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may > contain confidential and privileged information for the use of the > designated recipients. If you are not the intended recipient, (or > authorized to receive for the recipient) you are hereby notified that you > have received this communication in error and that any review, disclosure, > dissemination, distribution or copying of it or its contents is > prohibited. > If you have received this communication in error, please destroy all > copies > of this communication and any attachments and contact the sender by reply > e-mail or telephone (813) 281-0300. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 20 Date: Mon, 26 Sep 2005 19:59:41 -0700 From: Sam Vaughn Subject: [Histonet] section edges lifting To: histonet@lists.utsouthwestern.edu Message-ID: <1582e1c05092619596afc6ce1@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 I'm new to histonet and have a question about histo staining. I have 6 um paraffin sections of mouse knee joints on uncoated superfrost plus slides. I collect the sections from a distilled water bath and let them drain at least 30min before placing them at 37 degrees overnight before staining. After Safranin O/ Fast Green staining a very thin area around the edge at the edge of the tissue lifts off the slide rendering it impossible to visualize with the microscope. Of course this is the the articular surface which we are interested in! I've used the same slides in the past without any coating and it has worked fine. I'm in a new lab now and trying to get things running. Any help you could give me would be really appreciated! Thanks, Sam ------------------------------ Message: 21 Date: Tue, 27 Sep 2005 00:41:56 -0400 From: John Kiernan Subject: Re: [Histonet] Pink precipitate is monoformazan from BCIP/NBT reaction onISH? To: Ralph Puchalski Cc: histonet@lists.utsouthwestern.edu Message-ID: <4338CD94.F2BDE464@uwo.ca> Content-Type: text/plain; charset=us-ascii Dear Ralph, Your email is full of abbreviations and jargon. Am I right in thinking it's a question about localizing alkaline phosphatase activity by an indoxyl-tetrazolium method? If so, please provide a reference to the original publication and let us all know if you followed it exactly or made any changes. Part of your email suggests that the alkaline phosphatase activity is not endogenous but part of an amplification system used in in situ hybridization. The pH optima of endogenous and label alkaline phosphatases differ. The later paragraphs of your message indicate that you may not fully understand the significance of the mono- and diformazan products of reduction of nitro-BT. Both colours result from reduction of the tetrazolium salt, but you need controls to prove that the reduction was by bromochloroindoxyl and not by other reducing agents (such as diaphorases) in the tissue. Non-enzymatic reduction of tetrazolium salts by -SH has been a well known artifact {"nothing dehydrogenase") for 40-45 years. John A. Kiernan Anatomy Dept, UWO London, Canada. ___________________________________________________ Ralph Puchalski wrote: > > I am trying to figure out the origin of the pink precipitate in the > image I posted on http://www.histonet.org/site_images_frame.asp > > Please go to the image entitled: Pink Precipitate ver2.jpg. It is at > the top of the list on 9/26/05, posted by Ralph Puchalski. To see the > pink precipitate artifact, please open the image and set the scroll bars > on the bottom and right side of the image at their 1/2 way points. It > is ugly! > > I think this precipitate is the aggregation of the monoformazan > intermediate generated from NBT (after dephosphorylation of BCIP by > alkaline phosphatase) that is not completely reduced to diformazan, the > insoluble dark blue or black precipitate that labels cells expressing > target mRNAs in our in situ hybridization reactions. > > We don't know how the monoformazan adheres to the sections of tissue. > It appears to be non-covalent due to the tendency of the monoformazan to > migrate under the coverslip in the aqueous mounting medium that has yet > to dry, and form clumps or aggregates or pools as seen in the picture. > The monoformazan is soluble in ethanol so doesn't form pools or > aggregates. But as soon as it is exposed to an aqueous medium, it > precipitates. > > If we mount the post-ISH tissue sections with an organic based mounting > medium like UV-CureMount (Instrumedics), the monoformazan might cause > the entire section to turn to a brown tint as seen on > http://www.histonet.org/site_images_frame.asp > > Please go to the image Brown Tint 1.jpg. It is 4th image from the top > on 9/26, posted by Ralph Puchalski. The lower image is mounted with UV > CureMount, and the upper was with aqueous Hydro Matrix. There is no > pooling or precipitation of monoformazan with UV CureMount, but I think > it does cause the entire section to turn brown. > > Question: How do we eliminate this problem, which I believe is > monoformazan? If we reduce it fully using ascorbate, the section (later > mounted with HyrdoMatrix) turns dark blue or purple, the same color as > our ISH signal. We have tried washing off the monoformazan with 100% > ethanol prior to coverslipping, but only small amounts are removed. > 100% acetone also does not work effectively. > > Please let me know if you have any ideas that might help us eliminate > this problem. > > Thank you, > > Ralph > > Ralph Puchalski, Ph.D. > Manager, Process Engineering and Automation > Allen Institute for Brain Science > 551 N. 34th Street, Suite 200 > Seattle, WA 98103 > > ralphpu@alleninstitute.org > Tel: 206-548-7041 Fax: 206-548-7071 > www.brainatlas.org > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 33 **************************************** From gsmith <@t> confocal.com Tue Sep 27 09:10:37 2005 From: gsmith <@t> confocal.com (Glenn Smith) Date: Tue Sep 27 09:11:00 2005 Subject: [Histonet] Fluorescent Illumination Systems: Suggestions please? In-Reply-To: <200509270448.j8R4mL0F009876@mailstore2.execulink.net> Message-ID: <003801c5c36d$4104c390$1f05a8c0@confocal.com> Melissa, if you haven't already, I would suggest you also post the question on a microscopy listserve such as http://listserv.acsu.buffalo.edu/cgi-bin/wa?A0=confocal. Glenn Smith, P.Eng. 519.886.9013 x38 (office) 519.498.0614 (mobile) Biomedical Photometrics Inc/GeneFocus Widefield Confocal Scanning Instruments and Software -----Original Message----- Message: 3 Date: Mon, 26 Sep 2005 10:50:34 -0700 From: "Melissa Gonzalez" Subject: [Histonet] Fluorescent Illumination Systems: Suggestions please? To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello all, Sorry to re-post, but I only received one response to the following: I am currently in the market to upgrade our existing HB0 100 lamp house, which is very old, to several of new systems. Either the newest version of the HB0 100 from Zeiss, a mercury halide system called X-Cite from EXFO, or a xenon based burner called Lambda LS from Sutter Instruments. They are all comparable in price, but my main concern is increasing the brightness of my signal and evenly illuminating the field of view. I mostly look at DAPI, eGFP, FITC, and Alexa 594, which from spectral outputs, each one of these systems appears to have a different weakness in one of the above. On a plus, the Xcite rates their bulbs at 1500 hours, and the Lambda at 500-1000hrs. Does anyone out there have any input/comments/experience with these systems? (This would be hooked up to a Zeiss Axioplan) Please let me know if you have any info, as I have been unsuccessful so far with demos and need to place an order soon. Thanks a lot -I really appreciate it, Melissa ------------------------------ From bwhitaker <@t> brownpathology.com Tue Sep 27 09:22:49 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Tue Sep 27 09:18:33 2005 Subject: [Histonet] cpt codes for direct smears In-Reply-To: <200509270907.aa04099@MessagEX.iapc.net> Message-ID: <000001c5c36e$f20eeb10$3601a8c0@brownpathology.net> According to our billing company, we were being denied 88160 or 88161 if permanent sections were being done on the same specimen. We are billing an 88329 for those. What has everyone else's experience been? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 27, 2005 9:05 AM To: 'Barb Richmond'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cpt codes for direct smears We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Sep 27 10:02:24 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Sep 27 10:02:47 2005 Subject: cartilage lift up, mouse Re: [Histonet] section edges lifting In-Reply-To: <20050927131841.19876.qmail@web61224.mail.yahoo.com> References: <1582e1c05092619596afc6ce1@mail.gmail.com> <20050927131841.19876.qmail@web61224.mail.yahoo.com> Message-ID: <6.0.0.22.1.20050927085057.01b7bd18@gemini.msu.montana.edu> If one tries to flatten cartilage with a brush after it comes out of clearing agent, the cartilage often tears rather than flattening - been there, done that way too many times. It could be the cartilage is too dry to begin with, as it has a high content of water and overdehydration during processing may be a culprit. Check your processing schedule, in general, mouse knee joints will be totally processed in either 1 hour per station to 1 1/2 hours per station starting with 70%, 80% 95% X 2, 100% x 2, xylene x 2 and 3 paraffin changes at 60C using vacuum and pressure, automated processing is a huge help. If you have to make the cartilage lay down , it is better to do that after a water rinse and before alcohols/clearing agents in dehydration as solvents only make this tiny strip of articular cartilage brittle. It also helps to soak the trimmed bone block on ice with water on top before you start sectioning, just don't soak so long that the tissue swells out of the block and use the sharpest blade you have. The sections must be totally flat, compression free when picked up on Plus charge slides. After picking up on Plus charge (silane) we drain slides for 15 min or so, and then ALWAYS dry bone sections FLAT, at 37C to 40C to ensure the sections stay down. Flat drying in an oven is done over a period of days, 36 to 48 hours might be a ball park for time in terms of days, rather than overnight. This is for murine knees, paws, ankle joints, skulls, tibias, femurs and we do experience cartilage lift up. Gentle rinsing is a must. At 07:18 AM 9/27/2005, you wrote: >Check on the expirartion date of the slides. After some months they don't >work as they are supposed to. Slides should be drained/dried on edge (not >flat). A last resort would be to coverslip by hand, looking at the edges >(and even flattening them out with a hair brush before coverslipping). >Rene J. Buesa > >Sam Vaughn wrote: >I'm new to histonet and have a question about histo staining. >I have 6 um paraffin sections of mouse knee joints on uncoated superfrost >plus slides. I collect the sections from a distilled water bath and let them >drain at least 30min before placing them at 37 degrees overnight before >staining. After Safranin O/ Fast Green staining a very thin area around the >edge at the edge of the tissue lifts off the slide rendering it impossible >to visualize with the microscope. Of course this is the the articular >surface which we are interested in! I've used the same slides in the past >without any coating and it has worked fine. I'm in a new lab now and trying >to get things running. Any help you could give me would be really >appreciated! >Thanks, >Sam >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >--------------------------------- >Yahoo! for Good > Click here to donate to the Hurricane Katrina relief effort. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Sep 27 10:13:08 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Sep 27 10:13:25 2005 Subject: [Histonet] Re: Pink precipitate... from BCIP/NBT reaction on ISH? In-Reply-To: References: Message-ID: <6.0.0.22.1.20050927090322.01b6f6a0@gemini.msu.montana.edu> We have used DAKO's ready to use BCIP/NBT and always develop the color using a microscope. This particular product is fast, with color development in a shorter time, but that is the differences we experience as compared to Teri's longer time in the substrate. So while you are developing the color, be sure to monitor its progress with a microscope, pull your positive and look at it at different time increments, you can return it to the chromogen to proceed to color endpoint if not dark enough. We develop with this DAKO product on slides laying flat, but are careful to have microscopic monitoring, and do not experience ppt with this ready to use. Teri's suggestion is excellent and one that we use with AEC, the van der Loos way, so as to not get ppt on sections with that chromogen either. Vertical slides in a coplin jar and or slide mailer works too, but we do pull the positive for monitoring color development. Not only less time in chromogen but also check the concentration of your primary, if your color is coming up way too fast with too dark a ppt, you may need to adjust your primary concentration - BCIP/NBT is very sensitive, up there with DAB. Hopefully you did a dilution panel of the primary for optimal working concentration with this chromogen. At 07:37 AM 9/27/2005, you wrote: >Ralph, > >When we use BCIP/NBT, we do our color reaction development in the 37 >degree C incubator in the dark. We change the solution frequently >(every several hours, or when the solution turns pink). We do not lay >the slides flat for their incubation, instead we use the 5-slide plastic >mailers and have them standing upright. This keeps precipitate from >settling on to the glass. Many of the chromogens for Alk. Phos. tend to >form a precipitate, and the manufacturers don't recommend filtering. > >Your signal appears to be quite strong. Perhaps you can also decrease >the time in chromogen? > >Best of luck to you, > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From plucas <@t> biopath.org Tue Sep 27 10:06:15 2005 From: plucas <@t> biopath.org (Paula Lucas) Date: Tue Sep 27 10:15:01 2005 Subject: [Histonet] Holder for Sakura Staining Buckets Message-ID: <001f01c5c375$027f8a30$7c01a8c0@biopath.org> Our lab will stain two different methods on our Sakura DRS 2000 stainer. Pap staining and H&E staining. I want two separate staining buckets for some of the solutions, so buckets will be switched out and be replaced with another set. I'm searching for a product that will hold the buckets safely without the chance of it tipping over. Does anyone have any grand ideas? I appreciate it if you would pass on the information. Paula Lucas Bio-Path Medical Group From BRIAN.CHELACK <@t> usask.ca Tue Sep 27 10:46:25 2005 From: BRIAN.CHELACK <@t> usask.ca (Brian Chelack) Date: Tue Sep 27 10:48:44 2005 Subject: [Histonet] IHC background problems on chicken embryos Message-ID: <43396951.8AEE40B8@sask.usask.ca> Hello Katri; Your problem is related to the high levels of avidin in your tissues. Remember avidin is sourced from "chicken eggs"!!! In my veterinary IHC world I have pretty well given up on using avidin-biotin based IHC on avian samples, due to the excessive non-specific binding to various tissues, especially liver. My recommendation is to switch your detection system to a non-ABC based system such as Dako's Envision or even Peroxidase-anti-peroxidase. In general the amplification is a bit less using Envision or PAP, but the slides are much better signal to noise wise. regards Brian Chelack Prairie Diagnostic Services Saskatoon From m.wei <@t> biogenex.com Tue Sep 27 11:37:40 2005 From: m.wei <@t> biogenex.com (May Wei) Date: Tue Sep 27 11:38:29 2005 Subject: [Histonet] querry Message-ID: <37DC9F93CF7F864182D0463EF93D571B0B5086@ISLETON2.california.biogenex.com> Please visit http://www.ihcworld.com/_protocols/epitope_retrieval/frozen_section_ar.h tm Antigen Retrieval Method for Cryostat Frozen Tissue Sections Description: This is a simple method for antigen retrieval on aldehyde-fixed cryostat tissue sections or cultured cells. In many case, a brief 5 minutes pretreatment with 1% sodium dodecyl sulfate (SDS) produced a dramatic increase in staining intensity by immunohistochemistry and immunofluorescence. Solutions and Reagents: 1% Sodium Dodecyl Sulfate (SDS) in PBS: SDS --------------------------------------- 1 g 0.01M PBS (pH 7.4) ---------------- 100 ml Mix to dissolve. Procedure: Rinse sections three times for 5 min each in PBS. Cover sections with 1% SDS solution and incubate for 5 minutes at room temperature. Rinse sections three times for 5 min each in PBS. It is important to wash sections well, otherwise residual SDS will denature the antibodies subsequently applied to sections. Incubate sections in serum blocking solution. Incubate in the primary antibody and complete immunohistochemical staining steps as desired. References: Brown D, Lydon J, McLaughlin M, Stuart-Tilley A, Tyszkowski R, Alper S (1996) Antigen retrieval in cryostat tissue sections and cultured cells by treatment with sodium dodecyl sulfate (SDS). Histochem Cell Biol. 105(4):261-7. PubMed Abstract -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Arvind Sent: Tuesday, September 27, 2005 1:22 AM To: histonet@lists.utsouthwestern.edu Cc: arvind@nbrc.res.in Subject: [Histonet] querry can any one please tell me how to do antigen retrieval on soft tissue, what i do is i took cryosections on slide for human fetal brain tissue age upto 7 months (prenatal) sections are 40 micron thick, when i do HIER these sections are comming off the slides at some points, i use gelatin subbed slides for this please guide what to do in detail thanking in advance Arvind Singh Pundir National Brain Research Centre gurgaon, Haryana, INDIA arvind@nbrc.res.in _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcresor <@t> lcpath.com Tue Sep 27 11:51:47 2005 From: jcresor <@t> lcpath.com (Jennifer N. Cresor) Date: Tue Sep 27 11:52:15 2005 Subject: [Histonet] fatty tissue processing Message-ID: <200509271651.j8RGppt20673@plus34.host4u.net> Hello, We are currently in the process of setting up a processor specifically for processing fatty tissues. I would really like to get people's protocols on processing fatty tissues that have worked well for them. Thank you, Jennifer jcresor@icpath.com From shive003 <@t> umn.edu Tue Sep 27 11:55:53 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Tue Sep 27 11:56:14 2005 Subject: [Histonet] IHC background problems on chicken embryos References: <000b01c5c300$225661f0$6a9a9618@Katri> Message-ID: <002501c5c384$527054c0$41065486@auxs.umn.edu> I would second Brian Chelack's suggestion to use a NON-biotinylated IHC staining system with avian tissue. I use EnVision+/HRP exclusively (I stain for a few avian viruses here). I would also suggest adding 2% normal chicken serum per volume to the EnVision+/HRP reagent. It helps block nonspecific binding of the EnVision to endogenous chicken Igs and serum components. Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "Katri Tuomala" To: "HistoNet Server" Sent: Monday, September 26, 2005 8:09 PM Subject: [Histonet] IHC background problems on chicken embryos > Hi Histonetters, > A colleague of mine is trying to get VWF (Dakocytomation) and VEGF(R&D > Systems) working on chicken tissue. The antibodies are performing well on > human and rat tissue, but excessive background is the problem with the > chicken. Would the blocking serums (normal goat and normal rabbit > respectively) be the problem as well as the biotinylated secondary reagents > raised in goat and rabbit? My experience is strictly with human tissues, so > I am stumped. To my knowledge neither of these antibodies have been proven > to work with chicken, so maybe that is the problem. > Any advice from "chicken" people? > > Katri > > Katri Tuomala > Hamilton, Ontario, Canada > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From bmunk <@t> uwyo.edu Tue Sep 27 12:22:06 2005 From: bmunk <@t> uwyo.edu (Brandon Munk) Date: Tue Sep 27 12:22:36 2005 Subject: [Histonet] 1% gelatin/40% etOH mounting solution Message-ID: Histonetters, I am trying to get fixed tissue sections to stick onto slides better. I have tried subbed slides, plus slides (subbed and un-subbed) with only mediocre results. I was recently given a recipe for a 1% gelatin in 40% etOH mounting solution. This seems to work wonderfully on un-subbed plus slides, but I am having much difficulty in keeping the gelatin in solution long enough for it to be useful for mounting my sections. The recipe consists of heating distilled water (not over 60C) dissolving gelatin (enough for a 1% final concentration) and adding 80% etOH in two steps to the cooling gelatin solution, half soon after gelatin goes into solution and the other half sometime into the cooling of the solution. The recipe is vague since there is probably some voodoo that goes along with it. I have tried about 10 different variations on this recipe, none work consistently. Does anyone have some tips, pointers or advice to help me out, or even another recipe that may work better. Thanks in advance! Brandon Munk Dept. Zoology and Physiology University of Wyoming From jkiernan <@t> uwo.ca Tue Sep 27 12:18:14 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Tue Sep 27 12:28:37 2005 Subject: [Histonet] querry [SDS and IHC] References: <37DC9F93CF7F864182D0463EF93D571B0B5086@ISLETON2.california.biogenex.com> Message-ID: <43397ED6.8FAF7B6E@uwo.ca> That's a useful reference (Brown et al., 1996). Thanks for sharing it. I'm probably being a bit fussy, but treating frozen sections or cell cultures with a detergent isn't really antigen retrieval, despite the title of the paper. The SDS makes holes in the cell membranes, so that the big antibody molecules can get inside. Triton-X (a non-ionic detergent) is often used for this purpose on thick free-floating frozen sections of minimally formaldehyde-fixed brain. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ May Wei wrote: > > Please visit > http://www.ihcworld.com/_protocols/epitope_retrieval/frozen_section_ar.h > tm > > Antigen Retrieval Method for Cryostat Frozen Tissue Sections > > Description: This is a simple method for antigen retrieval on > aldehyde-fixed cryostat tissue sections or cultured cells. In many case, > a brief 5 minutes pretreatment with 1% sodium dodecyl sulfate (SDS) > produced a dramatic increase in staining intensity by > immunohistochemistry and immunofluorescence. > > Solutions and Reagents: > > 1% Sodium Dodecyl Sulfate (SDS) in PBS: > SDS --------------------------------------- 1 g > 0.01M PBS (pH 7.4) ---------------- 100 ml > Mix to dissolve. > > Procedure: > > Rinse sections three times for 5 min each in PBS. > Cover sections with 1% SDS solution and incubate for 5 minutes at room > temperature. > Rinse sections three times for 5 min each in PBS. It is important to > wash sections well, otherwise residual SDS will denature the antibodies > subsequently applied to sections. > Incubate sections in serum blocking solution. > Incubate in the primary antibody and complete immunohistochemical > staining steps as desired. > > References: > > Brown D, Lydon J, McLaughlin M, Stuart-Tilley A, Tyszkowski R, Alper S > (1996) Antigen retrieval in cryostat tissue sections and cultured cells > by treatment with sodium dodecyl sulfate (SDS). Histochem Cell Biol. > 105(4):261-7. PubMed Abstract > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Arvind > Sent: Tuesday, September 27, 2005 1:22 AM > To: histonet@lists.utsouthwestern.edu > Cc: arvind@nbrc.res.in > Subject: [Histonet] querry > > can any one please tell me how to do antigen retrieval on soft tissue, > what i do is i took cryosections on slide for human fetal brain tissue > age upto > 7 months (prenatal) sections are 40 micron thick, when i do HIER these > sections are comming off the slides at some points, i use gelatin > subbed slides for this please guide what to do in detail > > thanking in advance > > Arvind Singh Pundir > > National Brain Research Centre > gurgaon, Haryana, INDIA > arvind@nbrc.res.in > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Tue Sep 27 12:34:39 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Tue Sep 27 12:34:59 2005 Subject: [Histonet] fatty tissue processing Message-ID: <20050927173440.66575.qmail@web30403.mail.mud.yahoo.com> Hi Jennifer, I am sure you will get good suggestions for protocols from the many experienced histologists here. If you want any of them to wotk well, the tissue must be cut no thicker than 3 mm across its full face (thinner if possible) and these pieces should be given time for thourough fixation in formalin. Make sure your processor is not overloaded and solutions are changed regularly. Stephen From maria <@t> ski.org Tue Sep 27 12:51:34 2005 From: maria <@t> ski.org (Maria Mejia) Date: Tue Sep 27 12:51:56 2005 Subject: [Histonet] 1% gelatin/40% etOH mounting solution References: Message-ID: <433986A6.2000004@ski.org> Brandon, I have a few questions for you. Please, tell me what was the tissue fixed in? What's the tissue that your working with? How thick are these sections? Are these sections FFPE (formalin-fixed paraffin embedded)? And, finally what do you plan to do with the sections, i.e. IHC or other? Regards Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA, 94115 Email: maria@ski.org Phone: (415)-345-2185 Brandon Munk wrote: >Histonetters, > >I am trying to get fixed tissue sections to stick onto slides better. I have >tried subbed slides, plus slides (subbed and un-subbed) with only mediocre >results. I was recently given a recipe for a 1% gelatin in 40% etOH mounting >solution. This seems to work wonderfully on un-subbed plus slides, but I am >having much difficulty in keeping the gelatin in solution long enough for it >to be useful for mounting my sections. The recipe consists of heating >distilled water (not over 60C) dissolving gelatin (enough for a 1% final >concentration) and adding 80% etOH in two steps to the cooling gelatin >solution, half soon after gelatin goes into solution and the other half >sometime into the cooling of the solution. The recipe is vague since there >is probably some voodoo that goes along with it. I have tried about 10 >different variations on this recipe, none work consistently. Does anyone >have some tips, pointers or advice to help me out, or even another recipe >that may work better. Thanks in advance! > >Brandon Munk >Dept. Zoology and Physiology >University of Wyoming > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From dford <@t> deltapathology.com Tue Sep 27 13:17:09 2005 From: dford <@t> deltapathology.com (D. Ford) Date: Tue Sep 27 13:11:06 2005 Subject: [Histonet] Positions in Louisiana Message-ID: Delta Pathology, in Shreveport Louisiana, currently has three open histotech positions. If you have been displaced by the hurricanes and want to return to your home state or if you are looking to relocate please contact us for more information. Darlene Ford, B.S. CT, HT (ASCP) Technical Supervisor 2915 Missouri Avenue Shreveport, LA 71109 318-621-8820 phone 318-671-5922 fax From c.m.vanderloos <@t> amc.uva.nl Tue Sep 27 13:54:50 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Tue Sep 27 13:55:34 2005 Subject: [Histonet] RE: antigen retrieval for IHC Message-ID: <94534e93fa70.93fa7094534e@amc.uva.nl> Dear Patsy and others, Recently we did something similar with a tonsil that was cut in 10 pieces and formalin-fixed from 12 hours up to one year. Per fixation time three samples were taken for a tissue array. For example, our staining results showed that at least caspase-3 nicely stained the 1 year-fixed tissue sample. Ki67-staining "died" after 3 months of fixation. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Mon, 26 Sep 2005 09:58:46 -0600 From: "Patsy Ruegg" Subject: RE: antigen retrieval for IHC Shi mentions in his book on pg.10 ANTIGEN RESTORATION "Simply bathing deparaffinized sections in a cold 20% sucrose-saline solution could, over several days, restore a certain amount of immunoreactivity." In Introduction to IHC by Polak and Van Noorden on pg 24 3.8.1 "The simplest form of reversing the effects of formalin is to wash the tissue well before processing" I am about to test these statements, as I just received tissues for an IHC project that have been in 10% NBF for over one year. I washed the tissues in running tap h20 for 2 hrs., I will now process them into paraffin. I am planning as well to put some of the sections in 20% sucrose at 4dc for 2 days if I have trouble getting good IHC signal. I will keep you all posted on this experiment. These are samples of mouse mammory tissue. I will be testing them with cleaved caspase 3 ! and Ki67 From c.m.vanderloos <@t> amc.uva.nl Tue Sep 27 13:57:44 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Tue Sep 27 13:58:05 2005 Subject: [Histonet] RE: Low temp Antigen Retrieval for Bone Sections Message-ID: <943bbe941cd5.941cd5943bbe@amc.uva.nl> Dear Kim and others, We have been testing (not for bone unfortunately) the overnight antigen retrieval at 60-70C with both citrate pH6.0 and Tris-EDTA pH9.0 and compared the staining results with the short boiling method (15 min + 20 min cool-down). After overnight antigen retrieval using citrate6.0 the staining intensity was much less than with the boiling method. However, after overnight antigen retrieval using Tris-EDTA9.0 compared with the short boiling procedure the staining intensity was sometimes similar, sometimes less, sometimes even stronger. As ever, it varied per antigen. Interestingly, during the overnight procedure the "fatty tissue parts" stayed much better on the glass than after the short boiling procedure. Nearly the same was described by Junn Chavez and Tim Morken in their NSH-poster. Overnight antigen retrieval at 60-70C is to my opinion something worthwhile to investigate further. It's definitely interesting for those working with tissues that tends to fall off during antigen retrieval using the short boiling procedure. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Mon, 26 Sep 2005 11:34:06 -0700 (PDT) From: Kim Merriam Subject: [Histonet] Low temp Antigen Retrieval for Bone Sections To: Histonet Hello All, I will be performing some IHC on mouse femurs (about 12 or so different antibodies). In the past, I have had a lot of trouble keeping these sections on the slides during HIER. I did a quick search on the archives and there were lots of suggestions, but nothing definitive. I was reading an article about "low temp AR", overnight at 60C in TEG buffer, pH 9.0. Has anyone tried this method? Also - what does TEG buffer stand for? Any help would be greatly appreciated. Kim Kim Merriam Novartis Cambridge, MA From jkiernan <@t> uwo.ca Tue Sep 27 14:07:55 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Tue Sep 27 14:08:30 2005 Subject: [Histonet] 1% gelatin/40% etOH mounting solution References: Message-ID: <4339988B.CCE7317F@uwo.ca> Putting an adhesive on positively charged slides seems rather extravagant. The positive charge is there to attract the predominantly negatively charged carboxylate groups of the tissue's structural proteins. Covering the surface of the glass with a layer of gelatin or chrome-gelatin (subbing) will occlude the positive charges that you paid extra for. http://publish.uwo.ca/~jkiernan/adhesivs.htm for more information. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Brandon Munk wrote: > > Histonetters, > > I am trying to get fixed tissue sections to stick onto slides better. I have > tried subbed slides, plus slides (subbed and un-subbed) with only mediocre > results. I was recently given a recipe for a 1% gelatin in 40% etOH mounting > solution. This seems to work wonderfully on un-subbed plus slides, but I am > having much difficulty in keeping the gelatin in solution long enough for it > to be useful for mounting my sections. The recipe consists of heating > distilled water (not over 60C) dissolving gelatin (enough for a 1% final > concentration) and adding 80% etOH in two steps to the cooling gelatin > solution, half soon after gelatin goes into solution and the other half > sometime into the cooling of the solution. The recipe is vague since there > is probably some voodoo that goes along with it. I have tried about 10 > different variations on this recipe, none work consistently. Does anyone > have some tips, pointers or advice to help me out, or even another recipe > that may work better. Thanks in advance! > > Brandon Munk > Dept. Zoology and Physiology > University of Wyoming > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JPCOLEMA <@t> sentara.com Tue Sep 27 14:13:27 2005 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Tue Sep 27 14:14:13 2005 Subject: [Histonet] What's mellitin? Message-ID: I never heard of mellitin. If you mean melanin, there is a bleach method to remove it and compare to unbleached h&e slides and there is a Fontana Masson which is a melanin silver stain. >>> histonet-request@lists.utsouthwestern.edu 09/25/05 1:03 PM >>> Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. lab design (Melissa Jans) 2. RE: lab design (Jim Staruk) 3. unsubscribe (Anne Van Binsbergen) 4. unsubscribe (Anne Van Binsbergen) 5. RE: lab design (Bartlett, Jeanine) 6. Mellitin (TLCLab) ---------------------------------------------------------------------- Message: 1 Date: Sat, 24 Sep 2005 11:08:36 -0700 (PDT) From: Melissa Jans Subject: [Histonet] lab design To: histonet@lists.utsouthwestern.edu Message-ID: <20050924180837.32315.qmail@web54709.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 2 Date: Sat, 24 Sep 2005 20:59:22 -0400 From: "Jim Staruk" Subject: RE: [Histonet] lab design To: "'Melissa Jans'" , Message-ID: <0INC001DZLIAFRF3@vms046.mailsrvcs.net> Content-Type: text/plain; charset=us-ascii This is exactly what I did when I designed our new facilities. I built a flame-resistant room which is well-ventilated directly to the outside. The ventilation unit is adjustable for varying amounts of cubic feet per minute air exchange. This unit is always on. There is a special cabinet for flammable chemicals, acid-resistant bench tops for the tissue processors and multiple, adjustable shelves for slide drying in this room. The solid wood passage door is always closed. I also built a fume hood directly on the other side of one of these walls and placed a blower unit (from an old, charcoal bench-top fume hood) through the wall. This is where we coverslip. When the blower is on, it sucks away all of the xylene-laden air into the vented room which, in turn, is vented directly to the outside. There is never any xylene fumes in the lab. On our main homepage, there's a link to photos of our new lab. Here, you will see the ventilated room as well as the coverslipping fume hood on the other side of the ventilated room. Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Melissa Jans Sent: Saturday, September 24, 2005 2:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab design I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Sun, 25 Sep 2005 14:57:20 +0400 From: "Anne Van Binsbergen" Subject: [Histonet] unsubscribe To: , Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CC08@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" ------------------------------ Message: 4 Date: Sun, 25 Sep 2005 14:57:20 +0400 From: "Anne Van Binsbergen" Subject: [Histonet] unsubscribe To: , Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CC08@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" ------------------------------ Message: 5 Date: Sun, 25 Sep 2005 08:22:08 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] lab design To: "Melissa Jans" , Message-ID: Content-Type: text/plain; charset="utf-8" Hi Jan: Currently we have a separate room containing our tissue processors and embedding centers. We also have a large flammable storage cabinet in this room. We have a separate room with our alcohol/solvent recycler and there is a large flammable storage cabinet in there as well. We have 2 smaller flammable cabinets in routine lab space as well. We are moving into a brand-new lab building this fall. In the new lab we also will have separate rooms; one for the processors and embedding cetners and another for recycling. I do not think there is flammable storage in the new processing room (it's pretty small) but there is in the recycling room. I believe there will be at least one under-counter flammable storage in a main lab space as well. Jeanine Bartlett CDC, Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Melissa Jans Sent: Sat 9/24/2005 2:08 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] lab design I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Sun, 25 Sep 2005 17:03:01 +0200 From: TLCLab Subject: [Histonet] Mellitin To: histonet@lists.utsouthwestern.edu Message-ID: <4336BC25.9050801@free.fr> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hello, Is there any - strict - histochemical method to caracterise mellitin (meaning not an immunohistochemical method) ? Regards, Florent. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 31 **************************************** From JPCOLEMA <@t> sentara.com Tue Sep 27 14:15:38 2005 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Tue Sep 27 14:16:08 2005 Subject: [Histonet] Ah- mellitin Message-ID: Do you mean mellitin the bee venom component? >>> histonet-request@lists.utsouthwestern.edu 09/25/05 1:03 PM >>> Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. lab design (Melissa Jans) 2. RE: lab design (Jim Staruk) 3. unsubscribe (Anne Van Binsbergen) 4. unsubscribe (Anne Van Binsbergen) 5. RE: lab design (Bartlett, Jeanine) 6. Mellitin (TLCLab) ---------------------------------------------------------------------- Message: 1 Date: Sat, 24 Sep 2005 11:08:36 -0700 (PDT) From: Melissa Jans Subject: [Histonet] lab design To: histonet@lists.utsouthwestern.edu Message-ID: <20050924180837.32315.qmail@web54709.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 2 Date: Sat, 24 Sep 2005 20:59:22 -0400 From: "Jim Staruk" Subject: RE: [Histonet] lab design To: "'Melissa Jans'" , Message-ID: <0INC001DZLIAFRF3@vms046.mailsrvcs.net> Content-Type: text/plain; charset=us-ascii This is exactly what I did when I designed our new facilities. I built a flame-resistant room which is well-ventilated directly to the outside. The ventilation unit is adjustable for varying amounts of cubic feet per minute air exchange. This unit is always on. There is a special cabinet for flammable chemicals, acid-resistant bench tops for the tissue processors and multiple, adjustable shelves for slide drying in this room. The solid wood passage door is always closed. I also built a fume hood directly on the other side of one of these walls and placed a blower unit (from an old, charcoal bench-top fume hood) through the wall. This is where we coverslip. When the blower is on, it sucks away all of the xylene-laden air into the vented room which, in turn, is vented directly to the outside. There is never any xylene fumes in the lab. On our main homepage, there's a link to photos of our new lab. Here, you will see the ventilated room as well as the coverslipping fume hood on the other side of the ventilated room. Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Melissa Jans Sent: Saturday, September 24, 2005 2:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] lab design I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Sun, 25 Sep 2005 14:57:20 +0400 From: "Anne Van Binsbergen" Subject: [Histonet] unsubscribe To: , Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CC08@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" ------------------------------ Message: 4 Date: Sun, 25 Sep 2005 14:57:20 +0400 From: "Anne Van Binsbergen" Subject: [Histonet] unsubscribe To: , Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CC08@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" ------------------------------ Message: 5 Date: Sun, 25 Sep 2005 08:22:08 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] lab design To: "Melissa Jans" , Message-ID: Content-Type: text/plain; charset="utf-8" Hi Jan: Currently we have a separate room containing our tissue processors and embedding centers. We also have a large flammable storage cabinet in this room. We have a separate room with our alcohol/solvent recycler and there is a large flammable storage cabinet in there as well. We have 2 smaller flammable cabinets in routine lab space as well. We are moving into a brand-new lab building this fall. In the new lab we also will have separate rooms; one for the processors and embedding cetners and another for recycling. I do not think there is flammable storage in the new processing room (it's pretty small) but there is in the recycling room. I believe there will be at least one under-counter flammable storage in a main lab space as well. Jeanine Bartlett CDC, Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Melissa Jans Sent: Sat 9/24/2005 2:08 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] lab design I hope this email finds everyone well, I have the opportunity to do some redesigning of our laboratory space. I would like some feedback from some of the larger labs as to whether they have a separate room for their tissue processors and/or flammable storage. If so, how is it separate from the main laboratory working space...wall, door, both? Any feedback would be appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Sun, 25 Sep 2005 17:03:01 +0200 From: TLCLab Subject: [Histonet] Mellitin To: histonet@lists.utsouthwestern.edu Message-ID: <4336BC25.9050801@free.fr> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hello, Is there any - strict - histochemical method to caracterise mellitin (meaning not an immunohistochemical method) ? Regards, Florent. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 31 **************************************** From meligroc <@t> zgi.com Tue Sep 27 14:47:56 2005 From: meligroc <@t> zgi.com (CRME (Criss Meligro)) Date: Tue Sep 27 14:48:25 2005 Subject: [Histonet] Hydrogen Peroxide for IHC Message-ID: Just wondering if anyone has purchased 3% Hydrogen Peroxide in the 500 ml bottle from the local drug store at $1.00 +/- and used it for IHC instead of the $66 bottle from Ventana or similar company? Do you add tween? thanks....Criss From rcharles <@t> state.pa.us Tue Sep 27 14:55:58 2005 From: rcharles <@t> state.pa.us (Charles, Roger) Date: Tue Sep 27 14:56:27 2005 Subject: [Histonet] Hydrogen Peroxide for IHC Message-ID: <12E4E17FEF6EBE4BAE95BEB3CDCB5700025F117C@enhbgpri04.pa.lcl> We use store bought 3% hydrogen peroxide with no problems. Roger Charles Pa Veterinary Laboratory Harrisburg, PA. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of CRME (Criss Meligro) Sent: Tuesday, September 27, 2005 3:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hydrogen Peroxide for IHC Just wondering if anyone has purchased 3% Hydrogen Peroxide in the 500 ml bottle from the local drug store at $1.00 +/- and used it for IHC instead of the $66 bottle from Ventana or similar company? Do you add tween? thanks....Criss _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Linke_Noelle <@t> Allergan.com Tue Sep 27 14:58:22 2005 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Tue Sep 27 14:59:08 2005 Subject: [Histonet] Hydrogen Peroxide for IHC Message-ID: I go middle of the road.....I use the 500mL bottles from VWR at about $12 a bottle, and I don't add anything to it. Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of CRME (Criss Meligro) Sent: Tuesday, September 27, 2005 12:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hydrogen Peroxide for IHC Just wondering if anyone has purchased 3% Hydrogen Peroxide in the 500 ml bottle from the local drug store at $1.00 +/- and used it for IHC instead of the $66 bottle from Ventana or similar company? Do you add tween? thanks....Criss _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From japoteete <@t> saintfrancis.com Tue Sep 27 15:04:15 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Tue Sep 27 15:07:06 2005 Subject: [Histonet] Hydrogen Peroxide for IHC Message-ID: I don't trust drug store Hydrogen Peroxide. You never know how long it has been on the shelf or how long it had been sitting in a hot warehouse. Jacquie Poteete MT(ASCP)QIHC Lead technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: CRME (Criss Meligro) [mailto:meligroc@zgi.com] Sent: Tuesday, September 27, 2005 2:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hydrogen Peroxide for IHC Just wondering if anyone has purchased 3% Hydrogen Peroxide in the 500 ml bottle from the local drug store at $1.00 +/- and used it for IHC instead of the $66 bottle from Ventana or similar company? Do you add tween? thanks....Criss _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From gcallis <@t> montana.edu Tue Sep 27 15:33:19 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Sep 27 15:33:32 2005 Subject: [Histonet] 1% gelatin/40% etOH mounting solution In-Reply-To: References: Message-ID: <6.0.0.22.1.20050927142502.01b5a500@gemini.msu.montana.edu> Brandon, I have never heard of using alcohol with gelatin to make up a section adhesive. If you see ppt or gelatin coming out of solution, I would strongly suspect the gelatin is being "fixed" by the alcohol since gelatin is a protein and alcohol fixes proteins, what you see is probably precipitated (denatured) gelatin, not voodoo, just chemistry. Just an educated guess here. Chrome gelatin subbing solution: Dissolve 10 g gelatin in 1 liter WARM distilled water, add 1 gm chromium potassium sulfate, stire until dissolved, add a thymol crystal for preservation. Final concentration of gelatin is 1%. You can also make this a 0.5% solution if you wish. Use 100 bloom gelatin for most tissures. For bone, use 275 bloom gelatin made from pig collagen. Add 10 mls of this solution to 2 liter water bath or sub clean, uncoated microscope slides. Reduce background staining by dipping dry subbed slides in NBF 10 times, rinse well, air dry, store. Do NOT coat Plus Charge slides as these slides are already coated with silane to attract negatively charged tissue protein components. At 11:22 AM 9/27/2005, you wrote: >Histonetters, > >I am trying to get fixed tissue sections to stick onto slides better. I have >tried subbed slides, plus slides (subbed and un-subbed) with only mediocre >results. I was recently given a recipe for a 1% gelatin in 40% etOH mounting >solution. This seems to work wonderfully on un-subbed plus slides, but I am >having much difficulty in keeping the gelatin in solution long enough for it >to be useful for mounting my sections. The recipe consists of heating >distilled water (not over 60C) dissolving gelatin (enough for a 1% final >concentration) and adding 80% etOH in two steps to the cooling gelatin >solution, half soon after gelatin goes into solution and the other half >sometime into the cooling of the solution. The recipe is vague since there >is probably some voodoo that goes along with it. I have tried about 10 >different variations on this recipe, none work consistently. Does anyone >have some tips, pointers or advice to help me out, or even another recipe >that may work better. Thanks in advance! > >Brandon Munk >Dept. Zoology and Physiology >University of Wyoming > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From thomas.riddle <@t> hotmail.com Tue Sep 27 15:33:30 2005 From: thomas.riddle <@t> hotmail.com (Thomas Riddle) Date: Tue Sep 27 15:33:56 2005 Subject: [Histonet] Tissue Processor Message-ID: Hello Histonetters, our lab is currently looking into purchasing a Peloris Tissue Processor and I was wondering what the likes and dislikes for this product are befoer we make this big step! Please feel free to contact me directly... Your help is greatly appreciated! Tom _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From rjbuesa <@t> yahoo.com Tue Sep 27 16:09:41 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Sep 27 16:10:07 2005 Subject: [Histonet] Hydrogen Peroxide for IHC In-Reply-To: Message-ID: <20050927210941.44171.qmail@web61224.mail.yahoo.com> Our lab we always used 3% hydrogen peroxide manufactured by Fisher. We were not willing to pay for "brand" 3% H202 like Ventana. Rene J. "Poteete, Jacquie A." wrote: I don't trust drug store Hydrogen Peroxide. You never know how long it has been on the shelf or how long it had been sitting in a hot warehouse. Jacquie Poteete MT(ASCP)QIHC Lead technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: CRME (Criss Meligro) [mailto:meligroc@zgi.com] Sent: Tuesday, September 27, 2005 2:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hydrogen Peroxide for IHC Just wondering if anyone has purchased 3% Hydrogen Peroxide in the 500 ml bottle from the local drug store at $1.00 +/- and used it for IHC instead of the $66 bottle from Ventana or similar company? Do you add tween? thanks....Criss _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From MEeva <@t> mednet.ucla.edu Tue Sep 27 16:19:05 2005 From: MEeva <@t> mednet.ucla.edu (Eeva, Mervi) Date: Tue Sep 27 16:19:33 2005 Subject: [Histonet] Methyl Green Message-ID: <1304F6F98344D61190180002A51311C0085AB02B@medmail4.mednet.ucla.edu> Hi All, I tried Methyl Green counterstaining on my double stained immunohistochemistry slides and didn't get any visible counterstaining. Tissue was FFPE brain and otherwise I used mostly Vector reagents with DAB and AP-red detection. Any ideas what went wrong? - Mervi Eeva, UCLA ---------------------------------------------------------- IMPORTANT WARNING: This email (and any attachments) is only intended for the use of the person or entity to which it is addressed, and may contain information that is privileged and confidential. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Unauthorized redisclosure or failure to maintain confidentiality may subject you to federal and state penalties. If you are not the intended recipient, please immediately notify us by return email, and delete this message from your computer. ---------------------------------------------------------- From maria <@t> ski.org Tue Sep 27 17:08:47 2005 From: maria <@t> ski.org (Maria Mejia) Date: Tue Sep 27 17:09:02 2005 Subject: [Histonet] 1% gelatin/40% etOH mounting solution References: Message-ID: <4339C2EF.6050700@ski.org> Brandon, Taught yourself some histology techniques - good for your!!! I use gelatin sub slide that I do in-house. It works for me. I start with clean slides and then I wash them...again...myself with a soft brush & liquid-Nox. I wash well with plenty of tap H20 (running water - 30mins to 1hr.) Then rinse well with DH20. While cleaned slide are setting in DH20, I make the gelatin solution using Sigma 300 bloom (since your using brain sections at 40 ums thick). > cold distilled water - 500ml > 300 bloom gelatin 5gm Mix well w/stirring bar & heat - make sure that solution is not over heated above 60C (cause it will denature the protein in the gelatin). I keep the temp at about 58C. **If gelatin solution starts to get cold, just put back into oven to keep warm and use when ready. > add 0.2 gm of chromium potassium sulfate. Mix well. > add a few grains of thymol for preservation. Filter solution using #4 filter paper and store in oven at about the same temp - 58C. Can do single or double coat of gelatin on slides: > dip slides either using slide rack or can also do by hand each slide individually dip each slide 10 times and dry slides up-right. > once slides are completely dry store in clean slide boxes and date. I keep gelatin dip slides only for 6 months. **If still having problems with sections falling off, repeat the procedure twice, so there is a second coat of gelatin on each slide with drying time in-between. But, I'm sure this will work for you! If you get any staining background, I'd give Gayle's method of reducing background a try! I sure will. Hope's this helps. Maria bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 Brandon Munk wrote: >I am a behavioral ecologist learning histology for a research project >sectioning bird brains and staining with cresyl violet. Birds were >transcardially perfused with 4% paraformaldehyde in the field, brains >removed and post-fixed for about a week before being stored in PB until >sectioning. Cryoprotected in 20% sucrose/PB solution until brains sink, >embedding in OCT and sectioned on freezing stage microtome at 40 um taking >every fifth section. Mounted on plus slides and stained with cresyl violet >for volumetric measurements of brain regions. I spent much of my summer >practicing sectioning and staining on house sparrows to fine tune my >protocol which got me to the microtome (over cryostat) and non-subbed plus >slides seemingly working best (now obviously not the case!). > >Brandon > >P.S. I tried your cresyl violet staining protocol and it worked wonderfully, >thanks! > > > > >>From: Maria Mejia >>Date: Tue, 27 Sep 2005 10:51:34 -0700 >>To: Brandon Munk >>Cc: >>Subject: Re: [Histonet] 1% gelatin/40% etOH mounting solution >> >>Brandon, >> >>I have a few questions for you. Please, tell me what was the tissue >>fixed in? >>What's the tissue that your working with? How thick are these sections? Are >>these sections FFPE (formalin-fixed paraffin embedded)? And, finally what >>do you plan to do with the sections, i.e. IHC or other? >> >>Regards >> >>Maria Bartola Mejia >>Smith-Kettlewell Eye Research Institute >>San Francisco, CA, 94115 >>Email: maria@ski.org >>Phone: (415)-345-2185 >> >> >> >>Brandon Munk wrote: >> >> >> >>>Histonetters, >>> >>>I am trying to get fixed tissue sections to stick onto slides better. I have >>>tried subbed slides, plus slides (subbed and un-subbed) with only mediocre >>>results. I was recently given a recipe for a 1% gelatin in 40% etOH mounting >>>solution. This seems to work wonderfully on un-subbed plus slides, but I am >>>having much difficulty in keeping the gelatin in solution long enough for it >>>to be useful for mounting my sections. The recipe consists of heating >>>distilled water (not over 60C) dissolving gelatin (enough for a 1% final >>>concentration) and adding 80% etOH in two steps to the cooling gelatin >>>solution, half soon after gelatin goes into solution and the other half >>>sometime into the cooling of the solution. The recipe is vague since there >>>is probably some voodoo that goes along with it. I have tried about 10 >>>different variations on this recipe, none work consistently. Does anyone >>>have some tips, pointers or advice to help me out, or even another recipe >>>that may work better. Thanks in advance! >>> >>>Brandon Munk >>>Dept. Zoology and Physiology >>>University of Wyoming >>> >>> >>>_______________________________________________ >>>Histonet mailing list >>>Histonet@lists.utsouthwestern.edu >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >>> >>> >>> >> >> > > > From cfranci <@t> rigel.com Tue Sep 27 18:38:42 2005 From: cfranci <@t> rigel.com (Christian Franci) Date: Tue Sep 27 18:39:05 2005 Subject: [Histonet] processing skin and intestines Message-ID: <0d17748db6262fc12891e50715c9c973@rigel.com> Howdy folks! We've been asked by some scientists to process skin, cheek pouches and intestines from mice for FF/paraffin embedding for IHC. I'm just wondering if any of you happen to have a good protocol for doing so. I know that for rat tissues one tends to use longer processing times.. We just got a Citadel processing machine and I'd like to set up a program for this. thanks a bunch in advance! C From katri <@t> cogeco.ca Tue Sep 27 19:48:07 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Tue Sep 27 19:48:30 2005 Subject: [Histonet] IHC background problems on chicken embryos References: <000b01c5c300$225661f0$6a9a9618@Katri> <002501c5c384$527054c0$41065486@auxs.umn.edu> Message-ID: <00a401c5c3c6$4af29540$6a9a9618@Katri> I knew you guys would not let me down! Thank you for all excellent replies. I will convey these pearls of wisdom to my colleague... Katri ----- Original Message ----- From: "Jan Shivers" To: "Katri Tuomala" Cc: "histonet" Sent: Tuesday, September 27, 2005 12:55 PM Subject: Re: [Histonet] IHC background problems on chicken embryos >I would second Brian Chelack's suggestion to use a NON-biotinylated IHC > staining system with avian tissue. I use EnVision+/HRP exclusively (I > stain > for a few avian viruses here). I would also suggest adding 2% normal > chicken serum per volume to the EnVision+/HRP reagent. It helps block > nonspecific binding of the EnVision to endogenous chicken Igs and serum > components. > > Jan Shivers > U of MN Vet Diag Lab > > ----- Original Message ----- > From: "Katri Tuomala" > To: "HistoNet Server" > Sent: Monday, September 26, 2005 8:09 PM > Subject: [Histonet] IHC background problems on chicken embryos > > >> Hi Histonetters, >> A colleague of mine is trying to get VWF (Dakocytomation) and VEGF(R&D >> Systems) working on chicken tissue. The antibodies are performing well on >> human and rat tissue, but excessive background is the problem with the >> chicken. Would the blocking serums (normal goat and normal rabbit >> respectively) be the problem as well as the biotinylated secondary > reagents >> raised in goat and rabbit? My experience is strictly with human tissues, > so >> I am stumped. To my knowledge neither of these antibodies have been >> proven >> to work with chicken, so maybe that is the problem. >> Any advice from "chicken" people? >> >> Katri >> >> Katri Tuomala >> Hamilton, Ontario, Canada >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From WWmn916 <@t> aol.com Tue Sep 27 20:01:28 2005 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Tue Sep 27 20:01:48 2005 Subject: [Histonet] Frozen tissue washing from slide Message-ID: I'm hoping someone can give me some idea why frozen muscle sections would suddenly start washing away from slide while trying to do PAS w/wo stain? Other muscle panel stains do okay in staining process.....just the PAS w/wo is recently having problems. -We use superfrost plus slides -Sections are fixed in 10% formalin prior to stain -Most water rinses are DI. Only one tap water rinse step right after amylase digest. -Does periodic acid have to be room temp prior to staining? -Could Shiffs reagent be too cold? -does temperature of water rinses affect adherance of sections to slides? -We use a 10% acid water rinse after heme staining. I recall if acid water is made incorrectly, tissue sections could wash much like they do if acid water in H+E stain is too strong. Thanks for any thoughts and help. From M.J.Vonk <@t> lumc.nl Wed Sep 28 00:59:31 2005 From: M.J.Vonk <@t> lumc.nl (Vonk, M.J. (KNO)) Date: Wed Sep 28 00:59:53 2005 Subject: [Histonet] Holder for Sakura Staining Buckets Message-ID: Hello Paula, We had the same problem. We have chosen for a very simple but effective method. Our technical support made a simple basket holder out of wood. Now the surface of the staining basket is larger and it will not tip over. Just 4 pieces of wood is enough. make them in a square so the basket will fit it. Greetings, Marcel Vonk Leiden Netherlands From c.m.vanderloos <@t> amc.uva.nl Wed Sep 28 01:55:54 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Wed Sep 28 01:56:15 2005 Subject: [Histonet] RE: Methyl Green Message-ID: <950f24952f1d.952f1d950f24@amc.uva.nl> Hi Mervi, The brown-red color combination you mentioned allows a simple hematoxilin counterstain. I would advise you to work with a diluted hematoxilin solution and keep the time short. Check out the counterstaining result microscopically and if needed repeat.You don't want intensely blue nuclei as this will mask your double staining. If you really wish to counterstain with methylgreen soak your slides in 100 mM Na-acetate buffer pH 5.2 for 15 min. Don't wash. Cover sections for 5 min with 0.1% methylgreen (Sigma M6776) dissolved in the acetate buffer as above. Wash briefly with with distilled water and dry the slides completely at a hot plate. Mount organically with VectaMount. This procedure circumvents dehydration by alcohols, which isn't good for your red alk. phos. reaction product. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Tue, 27 Sep 2005 14:19:05 -0700 From: "Eeva, Mervi" Subject: [Histonet] Methyl Green To: histonet@lists.utsouthwestern.edu Hi All, I tried Methyl Green counterstaining on my double stained immunohistochemistry slides and didn't get any visible counterstaining. Tissue was FFPE brain and otherwise I used mostly Vector reagents with DAB and AP-red detection. Any ideas what went wrong? - Mervi Eeva, UCLA From L.Driessen <@t> orthop.umcn.nl Wed Sep 28 02:46:45 2005 From: L.Driessen <@t> orthop.umcn.nl (L.Driessen@orthop.umcn.nl) Date: Wed Sep 28 02:47:13 2005 Subject: [Histonet] Dissolving Alizarin complexon Message-ID: <2E2AC813A84E054C8E8E572131082BE2145510@umcnet14.umcn.nl> Hello, Is there anyone who is familiar with dissolving alizarin complexon for bonelabeling? We inject this fluorochrome in our testanimals to monitor new boneformation. Because of that we need to sterilize the solution. For this we push the solution through a 0,2 um millipore-filter. The problem is that the filter gets clogged very fast. We've tried prefiltering through a paperfilter, but that doesn't solve the problem. Does anyone have a a solution for this? Greetings, L?on Driessen, Orthopaedic Research Lab UMCN St. Radboud, Nijmegen, The Netherlands From Kemlo.Rogerson <@t> elht.nhs.uk Wed Sep 28 03:05:12 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Wed Sep 28 03:05:32 2005 Subject: [Histonet] fatty tissue processing Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD58B@elht-exch1.xelht.nhs.uk> This question is a regular; I don't have a protocol as I am a Manager/ Cytologist but from memory fatty tissues are best cut thin, well fixed, dehydrated and then put into a clearing agent/ degreasant. After the fat has been 'removed' return to dyhydration, then 'clear' again and impregnate with wax. You can use many degreasants but I used xylene, Mr. Lillie 'waxes' eloquently about such matters. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer N. Cresor Sent: Tuesday, September 27, 2005 5:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fatty tissue processing Hello, We are currently in the process of setting up a processor specifically for processing fatty tissues. I would really like to get people's protocols on processing fatty tissues that have worked well for them. Thank you, Jennifer jcresor@icpath.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cmalc <@t> unimelb.edu.au Wed Sep 28 03:33:11 2005 From: cmalc <@t> unimelb.edu.au (Cathy Malcontenti-Wilson) Date: Wed Sep 28 03:33:35 2005 Subject: [Histonet] Superfrost Plus slides Message-ID: <6.2.1.2.2.20050928182518.02658550@mail.staff.unimelb.edu.au> Dear All, OK so question regarding the superfrost slides. I use them for immunostaining and I remember a few years ago when cutting sections and orientating them in the waterbath, if I got it wrong well bad luck because the sections were well and truly stuck within a few seconds. But now I can orientate them no problem, they lift off easily. Even though they look like most of the sections are still attached, I am getting all this strange background immunostaining around the edges of the tissues and sometimes it looks as though something has been damaging the tissue around the edges as well. I remember a while ago there was some discussion on superfrost slides on histonet but I cant remember what the conclusion was. Has anyone noticed any changes with the superfrost slides, or do they recommend one brand over another? We use Menzel brand and have tried two suppliers, and both the superfrost and the superfrost plus slides. Thanks in advance. Cathy From lpwenk <@t> sbcglobal.net Wed Sep 28 03:53:36 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Wed Sep 28 03:54:11 2005 Subject: [Histonet] Hydrogen Peroxide for IHC In-Reply-To: <20050927210941.44171.qmail@web61224.mail.yahoo.com> Message-ID: <1084225373-373221435@pathology.swmed.edu> We use "store bought". However, we buy it through our hospital pharmacy, so it comes in fresh and cheaper than off the drug store shelf. Plus, we look at the expiration date (which is always a year down the road). Peggy Wenk.HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, September 27, 2005 5:10 PM To: Poteete, Jacquie A.; 'CRME (Criss Meligro)'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Hydrogen Peroxide for IHC Our lab we always used 3% hydrogen peroxide manufactured by Fisher. We were not willing to pay for "brand" 3% H202 like Ventana. Rene J. "Poteete, Jacquie A." wrote: I don't trust drug store Hydrogen Peroxide. You never know how long it has been on the shelf or how long it had been sitting in a hot warehouse. Jacquie Poteete MT(ASCP)QIHC Lead technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: CRME (Criss Meligro) [mailto:meligroc@zgi.com] Sent: Tuesday, September 27, 2005 2:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hydrogen Peroxide for IHC Just wondering if anyone has purchased 3% Hydrogen Peroxide in the 500 ml bottle from the local drug store at $1.00 +/- and used it for IHC instead of the $66 bottle from Ventana or similar company? Do you add tween? thanks....Criss _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ree3 <@t> leicester.ac.uk Wed Sep 28 04:59:03 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Wed Sep 28 04:59:32 2005 Subject: [Histonet] RE: haemoxygenase I or II Message-ID: Wanted antibodies to the above that work on mouse paraffin processed tissues...thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K..... From stema_cba <@t> yahoo.it Wed Sep 28 07:47:23 2005 From: stema_cba <@t> yahoo.it (Stefano Mantero) Date: Wed Sep 28 07:47:40 2005 Subject: [Histonet] SEM on mouse tissue Message-ID: <20050928124723.41442.qmail@web25903.mail.ukl.yahoo.com> I need Histo help: I?m looking for a protocol for a Scanning Electron Microscopy sample preparation on embryonal mouse tissue. Please help ! Thanks in advance Stefano --------------------------------- Yahoo! Mail: gratis 1GB per i messaggi, antispam, antivirus, POP3 From Jeffery-Smith <@t> ouhsc.edu Wed Sep 28 09:07:15 2005 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Wed Sep 28 09:09:58 2005 Subject: [Histonet] Superfrost Plus slides Message-ID: We are using VWR Superfrost Plus. We do hundreds of Immunos per week and have had no such problems. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Cathy Malcontenti-Wilson Sent: Wed 9/28/2005 3:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Superfrost Plus slides Dear All, OK so question regarding the superfrost slides. I use them for immunostaining and I remember a few years ago when cutting sections and orientating them in the waterbath, if I got it wrong well bad luck because the sections were well and truly stuck within a few seconds. But now I can orientate them no problem, they lift off easily. Even though they look like most of the sections are still attached, I am getting all this strange background immunostaining around the edges of the tissues and sometimes it looks as though something has been damaging the tissue around the edges as well. I remember a while ago there was some discussion on superfrost slides on histonet but I cant remember what the conclusion was. Has anyone noticed any changes with the superfrost slides, or do they recommend one brand over another? We use Menzel brand and have tried two suppliers, and both the superfrost and the superfrost plus slides. Thanks in advance. Cathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Wed Sep 28 09:24:09 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Wed Sep 28 09:24:42 2005 Subject: [Histonet] Superfrost Plus slides Message-ID: <092820051424.17323.433AA7890005BBA8000043AB2207001641CECE030E9D0C9A03@comcast.net> Cathy, You should also be sure you are roatating your slides so the oldest lot numbers are used first. The coatings are not permenant and can lose surface attraction over time and with exposure to light, heat and air. The expiration date is from the time they are manufactured, not when they arrive to you. Be sure your supplier is sending you fresh slides that have not been sitting in a warehouse for months with varying temperatures and conditions. Hope this helps. Pam Marcum -------------- Original message -------------- > We are using VWR Superfrost Plus. We do hundreds of Immunos per week and have > had no such problems. > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Cathy > Malcontenti-Wilson > Sent: Wed 9/28/2005 3:33 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Superfrost Plus slides > > > > Dear All, > OK so question regarding the superfrost slides. > I use them for immunostaining and I remember a few years ago when cutting > sections and orientating them in the waterbath, if I got it wrong well bad > luck because the sections were well and truly stuck within a few seconds. > But now I can orientate them no problem, they lift off easily. > Even though they look like most of the sections are still attached, I am > getting all this strange background immunostaining around the edges of the > tissues and sometimes it looks as though something has been damaging the > tissue around the edges as well. I remember a while ago there was some > discussion on superfrost slides on histonet but I cant remember what the > conclusion was. > Has anyone noticed any changes with the superfrost slides, or do they > recommend one brand over another? > We use Menzel brand and have tried two suppliers, and both the superfrost > and the superfrost plus slides. > Thanks in advance. > Cathy > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-aventis.com Wed Sep 28 09:41:56 2005 From: Stephen.Eyres <@t> sanofi-aventis.com (Stephen.Eyres@sanofi-aventis.com) Date: Wed Sep 28 09:42:24 2005 Subject: [Histonet] Source of DIFCO trypsin in the UK Message-ID: Hi, Could someone give me a source for DIFCO trypsin (0152 or 21530-product code change???)? Many thanks Steve ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From gcallis <@t> montana.edu Wed Sep 28 10:00:08 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Sep 28 10:00:23 2005 Subject: [Histonet] processing skin and intestines In-Reply-To: <0d17748db6262fc12891e50715c9c973@rigel.com> References: <0d17748db6262fc12891e50715c9c973@rigel.com> Message-ID: <6.0.0.22.1.20050928085212.01b7f430@gemini.msu.montana.edu> Isn't the Citadel a carousel (sp?) style with vacuum on last station of paraffin? I am not totally familiar with this processor. With our VIP (vacuum/pressure throughout), 45 minutes per station of dehyrant, clearing and paraffin (60C) at most but we get superb exchange of solvents with VIP processing, and we never add heat to dehydration/clearing steps - dries out the tissue even more. For rat, I would do 1 hour on VIP. If you do not have vacuum and pressure throughout for each station, then mouse tissue will survive 1 hour timing on a carousel style nicely. Keep in mind animal are less fatty than human, and can easily be overdehydrated, dried out. At 05:38 PM 9/27/2005, you wrote: >Howdy folks! >We've been asked by some scientists to process skin, cheek pouches and >intestines from mice for FF/paraffin embedding for IHC. >I'm just wondering if any of you happen to have a good protocol for doing >so. I know that for rat tissues one tends to use longer processing times.. >We just got a Citadel processing machine and I'd like to set up a program >for this. >thanks a bunch in advance! > >C Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From DeBrosse_Beatrice <@t> Allergan.com Wed Sep 28 10:01:23 2005 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Wed Sep 28 10:02:12 2005 Subject: [Histonet] Frozen tissue washing from slide Message-ID: How long do you air dry the slides? Have you tried to fix the slides in acetone? Is the 10% formalin cold when you fix the slides? How thick are the sections? I really don't see a problem with the Periodic acid, the Schiffs or the water rinses, but I believe your acid solution is too concentrated after your counterstaining. Have you tried 1% acid/alcohol (70% alcohol)? Sincerely, Beatrice DeBrosse-Serra BS, HT(ASCP) Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of WWmn916@aol.com Sent: Tuesday, September 27, 2005 6:01 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Frozen tissue washing from slide I'm hoping someone can give me some idea why frozen muscle sections would suddenly start washing away from slide while trying to do PAS w/wo stain? Other muscle panel stains do okay in staining process.....just the PAS w/wo is recently having problems. -We use superfrost plus slides -Sections are fixed in 10% formalin prior to stain -Most water rinses are DI. Only one tap water rinse step right after amylase digest. -Does periodic acid have to be room temp prior to staining? -Could Shiffs reagent be too cold? -does temperature of water rinses affect adherance of sections to slides? -We use a 10% acid water rinse after heme staining. I recall if acid water is made incorrectly, tissue sections could wash much like they do if acid water in H+E stain is too strong. Thanks for any thoughts and help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Sep 28 10:08:59 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Sep 28 10:09:14 2005 Subject: [Histonet] Dissolving Alizarin complexon In-Reply-To: <2E2AC813A84E054C8E8E572131082BE2145510@umcnet14.umcn.nl> References: <2E2AC813A84E054C8E8E572131082BE2145510@umcnet14.umcn.nl> Message-ID: <6.0.0.22.1.20050928090319.01b71a78@gemini.msu.montana.edu> Have you tried a larger pore size, 0.45 um.or larger. One could always get the big particles first, then refilter again through smaller pore size if you felt it necessary. I have had to do this with normal serums before when 0.22um clogs up. Possibly, you could try centrifuging the stock solution, decant off the top what you need and microfilter that portion just to eliminate the biggest particles before you go to microfilter. Just suggestions here At 01:46 AM 9/28/2005, you wrote: >Hello, > >Is there anyone who is familiar with dissolving alizarin complexon for >bonelabeling? >We inject this fluorochrome in our testanimals to monitor new >boneformation. Because of that we need to sterilize the solution. For this >we push the solution through a 0,2 um millipore-filter. The problem is >that the filter gets clogged very fast. We've tried prefiltering through a >paperfilter, but that doesn't solve the problem. Does anyone have a a >solution for this? > >Greetings, > >L?on Driessen, >Orthopaedic Research Lab >UMCN St. Radboud, Nijmegen, >The Netherlands > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From anh2006 <@t> med.cornell.edu Wed Sep 28 10:29:57 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Wed Sep 28 10:25:49 2005 Subject: [Histonet] Pap Pen Blues Message-ID: Hello All, I have used Zymed Mini Pap pens for years now as through trials and tribulations and talking with coworkers we agreed it was the best out there. The pen rarely came up or leaked and gave beautiful results. Even withstood heat retrieval in DAKO Target Retrieval Solution with no problems. The stuff was indestructible!! This is until recently ... I have noticed with the past several orders that the formula has changed. The pap pen "ink" is now a blue green in color (used to be more generic yellow/beige) and it does not stay on well at all. I end up going home in a state of frustration every day as I pap pen hundreds of slides with little success for the entirety of my experiment. Arggggh! So I have two questions: (1) Has anyone else noticed this change in formula over the past year or so? (It may have been longer as I was working with a stock of pap pens before that) (2) Who makes your favorite pap pen? THANKS! Andrea -- From gcallis <@t> montana.edu Wed Sep 28 10:29:10 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Sep 28 10:29:32 2005 Subject: [Histonet] Superfrost Plus slides In-Reply-To: <6.2.1.2.2.20050928182518.02658550@mail.staff.unimelb.edu.a u> References: <6.2.1.2.2.20050928182518.02658550@mail.staff.unimelb.edu.au> Message-ID: <6.0.0.22.1.20050928090945.01b54cf0@gemini.msu.montana.edu> Cathy, A couple of things, for reorientation of sections, we use the Erie Polysine slides (poly l lysine coating) that lets us manipulate sections on slide at waterbath. Plus Charge are silane and once the section is attracted to slide surface, you are done! Do you use Tween or a detergent in your buffers/diluents before chromogen application and throughout the staining procedure? If not what you may be experiencing is hydrophobic bonding interactions between tissue and reagent proteins (BSA, serums) and ionic interaction of chromogen to slide surface, which the detergent will eliminate giving cleaner results. 0.05% Tween 20 is usually adequate to eliminate this problem. We have used Erie's SuperFrost Plus Charge, their trademark and also Polysine for section manipulation problems for years, and love them. We have used Plus charge slides stored long past expiration date without any problems with IHC or other routine staining. What other manufacturers do for coating slides may vary from Erie products, one can only try them out to see if they work well. As for tissue section damage, can you describe in further detail please? I doubt this is caused by the slide coatings but some other problem. At 02:33 AM 9/28/2005, you wrote: >Dear All, >OK so question regarding the superfrost slides. >I use them for immunostaining and I remember a few years ago when cutting >sections and orientating them in the waterbath, if I got it wrong well bad >luck because the sections were well and truly stuck within a few seconds. >But now I can orientate them no problem, they lift off easily. >Even though they look like most of the sections are still attached, I am >getting all this strange background immunostaining around the edges of the >tissues and sometimes it looks as though something has been damaging the >tissue around the edges as well. I remember a while ago there was some >discussion on superfrost slides on histonet but I cant remember what the >conclusion was. >Has anyone noticed any changes with the superfrost slides, or do they >recommend one brand over another? >We use Menzel brand and have tried two suppliers, and both the superfrost >and the superfrost plus slides. >Thanks in advance. >Cathy > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Sep 28 10:46:26 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Sep 28 10:46:45 2005 Subject: PAS staining RE: [Histonet] Frozen tissue washing from slide In-Reply-To: References: Message-ID: <6.0.0.22.1.20050928093504.01b8d3b8@gemini.msu.montana.edu> With frozen sections, a gentle rinse is advisable. We cut frozen sections destined for routine stains and go directly to room temperature NBF - air drying is not really necessary for routine stains but is for immunostaiing. Let your frozen sections fix longer than 10 minutes to ensure good adherance to slide and fixed proteins. You can proceed with a PAS stain just as you do a paraffin section, at room temperature. Is the 10% acid glacial acetic or hydrochloric acid? You did not specify which one. If you do acid/alcohol rinse, 0.5 to 1% HCl in 70% for regressive or Harris hematoxylin , or if you use acetic acid, 4% in water is sufficient with progressive Gill type hematoxylin. >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >WWmn916@aol.com >Sent: Tuesday, September 27, 2005 6:01 PM >To: histonet@pathology.swmed.edu >Subject: [Histonet] Frozen tissue washing from slide > >I'm hoping someone can give me some idea why frozen muscle sections >would >suddenly start washing away from slide while trying to do PAS w/wo >stain? >Other muscle panel stains do okay in staining process.....just the PAS >w/wo is >recently having problems. >-We use superfrost plus slides >-Sections are fixed in 10% formalin prior to stain >-Most water rinses are DI. Only one tap water rinse step right after >amylase digest. >-Does periodic acid have to be room temp prior to staining? >-Could Shiffs reagent be too cold? >-does temperature of water rinses affect adherance of sections to >slides? >-We use a 10% acid water rinse after heme staining. I recall if acid >water >is made incorrectly, tissue sections could wash much like they do if >acid >water in H+E stain is too strong. > >Thanks for any thoughts and help. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Sep 28 11:04:23 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Sep 28 11:04:35 2005 Subject: [Histonet] Pap Pen Blues In-Reply-To: References: Message-ID: <6.0.0.22.1.20050928095641.01b98148@gemini.msu.montana.edu> Andrea, We now use Vectors ImmEdge pens - doesn't come up, and has a hint of color. Cheaper, two to a set. Can remark a slide when wet, although we do blot away moisture, but the pen goo does not lift nor flow across a section if it contacts buffer - been there and have experienced the "Pap pen oil slick syndrome" that ruins IHC when section gets goo-coated. To use ImmEdge, be sure to shake pens very vigorously to redistribute the goo (per their instructions). We vortex them hard to do that job. Maybe the pens you have now need a good vortexing - worth a try for pricey little pens. Pressing the point to start goo is still something that requires gentle tough, but I have not had too many problems with that. Flow is generally excellent, right amount. At 09:29 AM 9/28/2005, you wrote: >Hello All, > >I have used Zymed Mini Pap pens for years now as through trials and >tribulations and talking with coworkers we agreed it was the best out >there. The pen rarely came up or leaked and gave beautiful results. Even >withstood heat retrieval in DAKO Target Retrieval Solution with no >problems. The stuff was indestructible!! > >This is until recently ... I have noticed with the past several orders >that the formula has changed. The pap pen "ink" is now a blue green in >color (used to be more generic yellow/beige) and it does not stay on well >at all. I end up going home in a state of frustration every day as I pap >pen hundreds of slides with little success for the entirety of my >experiment. Arggggh! > >So I have two questions: > >(1) Has anyone else noticed this change in formula over the past year or >so? (It may have been longer as I was working with a stock of pap pens >before that) >(2) Who makes your favorite pap pen? > >THANKS! >Andrea >-- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From hstevens <@t> ucsd.edu Wed Sep 28 11:20:45 2005 From: hstevens <@t> ucsd.edu (hstevens@ucsd.edu) Date: Wed Sep 28 11:21:06 2005 Subject: [Histonet] plastic sections Message-ID: <32021.67.113.110.37.1127924445.squirrel@acs-webmail.ucsd.edu> Hi, I am looking for a good inexpensive lab that can process mouse femurs (at present in alcohol) to obtain plastic embedded midshaft sections (30 um ish). Does anyone have any recommendations for the San Diego area or beyond? Thanks Hazel Stevens From TJJ <@t> Stowers-Institute.org Wed Sep 28 11:53:22 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Sep 28 11:54:17 2005 Subject: [Histonet] Use of methanol and DMSO in Whole mount immunostaining Message-ID: Ref: GENES & DEVELOPMENT 9:2509-2522, Misexpression of Hoxa-13 induces cartilage homeotic transformation and changes cell adhesiveness in chick limb buds, Yokouchi, et al In this study, chick embryos were dissected and fixed in methanol/DMSO (4:1) at 4 degrees C overnight, followed by peroxidase blocking in 5% hydrogen peroxide in methanol/DMSO (4:1). I am unfamiliar with using methanol/DMSO as a tissue fixative for whole mount studies. This can obviously be quite helpful for finding protein targets that are masked or made unrecognizable using aldehyde fixatives. What effect would the DMSO have on the samples prior to immunostaining? Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From lori.langiano <@t> medtronic.com Wed Sep 28 12:04:52 2005 From: lori.langiano <@t> medtronic.com (Langiano, Lori, MS, HT) Date: Wed Sep 28 12:05:10 2005 Subject: [Histonet] Soft tissue digestion Message-ID: <32D2979CA2119041B2A84241FCD860FD1C4E43@STSM1BMSGM01> Has anyone had experience with digesting soft tissue away from an implant? I need to completely remove the tissue from an implant. Thanks in advance for any suggestions. Lori From andromeda_tm <@t> libero.it Wed Sep 28 12:12:44 2005 From: andromeda_tm <@t> libero.it (Massimo) Date: Wed Sep 28 12:13:20 2005 Subject: [Histonet] Mayer's Haemalum reactive Message-ID: Dear Histonetters, I'd like to apologize if my question will sound too basic. I have stained a tissue with Mayer's Haemalum reactive. After I have put the specimen in water at a Ph a bit over 7, I would be expected a nuclear colour change from red "bordeaux" to blue. But it didn't happen. I fixed the specimen with Bouin's liquid and I clarified with Histoclear. I wonder why. Have you seen a behaviour of this kind? Thanks in advance for your help, Best Regards, Massimo From mweirauch <@t> crittenton.com Wed Sep 28 12:18:02 2005 From: mweirauch <@t> crittenton.com (Maray Weirauch) Date: Wed Sep 28 12:18:32 2005 Subject: [Histonet] cpt codes for direct smears Message-ID: We have also been denied payment when using 88160 or 88161. The Centers for Medicare and Medicaid Services (CMS) have taken the stand that cytopathology codes 88160, 88161 and 88162 can not be reported for the same patient on the same date of service with codes 88304 to 88309, 'Level III through VI - Surgical pathology, gross and microscopic examination.' CMS said it considers cytopathology smears prepared from surgical pathology specimens as duplicate procedures that should not be reported separately. They will allow the use of modifier -59 for bypass if the cytopathology smears are performed on separate specimens and independent diagnoses are rendered. That scenario never seems to apply to our situation, so we also bill for a Pathology consultation during surgery, 88329. >>> "Bonnie Whitaker" 9/27/2005 10:22:49 AM >>> According to our billing company, we were being denied 88160 or 88161 if permanent sections were being done on the same specimen. We are billing an 88329 for those. What has everyone else's experience been? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 27, 2005 9:05 AM To: 'Barb Richmond'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cpt codes for direct smears We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Sep 28 12:22:12 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Sep 28 12:22:25 2005 Subject: Enzyme method for Re: [Histonet] Soft tissue digestion In-Reply-To: <32D2979CA2119041B2A84241FCD860FD1C4E43@STSM1BMSGM01> References: <32D2979CA2119041B2A84241FCD860FD1C4E43@STSM1BMSGM01> Message-ID: <6.0.0.22.1.20050928111511.01b49278@gemini.msu.montana.edu> I used to digest soft tissue off skulls, long bones for museum teaching pieces using an enzyme detergent. Make up 10 to 20% BIZ detergent in water, heat to 80C in waterbath, and put bones, whatever in this for several hours. It will even remove fixed tissue (if you rinse it well with water before hand as NBF could inactivate the enzyme a bit. Do NOT boil. Rinse well with water afterwards, and soft tissue will release from bone, whatever. Beware, it will eat cartilage off bones. You can repeat until the implant is clean. Try to manipulate soft tissues off with gloved hands, or soft brushes. Best part of all this, is isn't smelly, and bones come out perfectly clean and white. I have a reference for this from J Microscopy, many moons ago. At 11:04 AM 9/28/2005, you wrote: >Has anyone had experience with digesting soft tissue away from an >implant? I need to completely remove the tissue from an implant. Thanks >in advance for any suggestions. >Lori >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From ROrr <@t> enh.org Wed Sep 28 13:13:00 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Wed Sep 28 13:13:22 2005 Subject: [Histonet] RE: Supervisor-Anatomic Pathology Message-ID: HI everyone, Anyone interested? It's a great place to work! Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- From: Wagner, Nikole Sent: Wednesday, September 28, 2005 8:29 AM To: Orr, Rebecca Subject: Supervisor-Anatomic Pathology Evanston Northwestern Healthcare is seeking a Laboratory Supervisor for our Anatomic Pathology Department at Evanston Hospital. The position requires HTL (ASCP) certification and 3-5 years of supervisory experience in a Histology Laboratory. Interested individuals can submit resumes to nwagner@enh.org or apply on-line at www.enh.org/careers. Nikole Wagner Sr. Human Resources Associate Evanston Northwestern Healthcare Phone. 847-570-1484 Fax. 847-570-1903 From Linke_Noelle <@t> Allergan.com Wed Sep 28 14:20:10 2005 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Wed Sep 28 14:19:57 2005 Subject: [Histonet] for anyone in pharmaceutical, contract labs, etc Message-ID: Hi All, I have a question regarding necropsy. Do you typically have one organ weigher, or does each individual do their own weighing? How many computers do you have set up in your necropsy rooms? Thank you!! Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 From TJJ <@t> Stowers-Institute.org Wed Sep 28 15:08:30 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Sep 28 15:09:11 2005 Subject: [Histonet] Re: Pap Pen Blues Message-ID: For What It's Worth (FWIW), I pretty well despise Pap pens. And I dearly LOVE Gayle's technical term (goo) for the stuff inside them. They're expensive, but see if you can get various companies to send you one to try. We have used them from Dako, Vector, and Biocare. We don't use them any more. We have the coverplates from Shandon and that works well for our hand-staining. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 Andrea, We now use Vectors ImmEdge pens - doesn't come up, and has a hint of color. Cheaper, two to a set. Can remark a slide when wet, although we do blot away moisture, but the pen goo does not lift nor flow across a section if it contacts buffer - been there and have experienced the "Pap pen oil slick syndrome" that ruins IHC when section gets goo-coated. To use ImmEdge, be sure to shake pens very vigorously to redistribute the goo (per their instructions). We vortex them hard to do that job. Maybe the pens you have now need a good vortexing - worth a try for pricey little pens. Pressing the point to start goo is still something that requires gentle tough, but I have not had too many problems with that. Flow is generally excellent, right amount. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) At 09:29 AM 9/28/2005, you wrote: >Hello All, > >I have used Zymed Mini Pap pens for years now as through trials and >tribulations and talking with coworkers we agreed it was the best out >there. The pen rarely came up or leaked and gave beautiful results. Even >withstood heat retrieval in DAKO Target Retrieval Solution with no >problems. The stuff was indestructible!! > >This is until recently ... I have noticed with the past several orders >that the formula has changed. The pap pen "ink" is now a blue green in >color (used to be more generic yellow/beige) and it does not stay on well >at all. I end up going home in a state of frustration every day as I pap >pen hundreds of slides with little success for the entirety of my >experiment. Arggggh! > >So I have two questions: > >(1) Has anyone else noticed this change in formula over the past year >or >so? (It may have been longer as I was working with a stock of pap pens >before that) >(2) Who makes your favorite pap pen? > >THANKS! >Andrea >-- > >________________________________ From anh2006 <@t> med.cornell.edu Wed Sep 28 15:27:25 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Wed Sep 28 15:23:09 2005 Subject: [Histonet] Re: Pap Pen Blues In-Reply-To: References: Message-ID: Hi Teri, Thanks for the reply. Coverplates are great except for when, like in my case, you have 3-6 sections per slide each to be stained with a different antibody or dilution thereof! Thanks for the info, Andrea >For What It's Worth (FWIW), I pretty well despise Pap pens. And I >dearly LOVE Gayle's technical term (goo) for the stuff inside them. > >They're expensive, but see if you can get various companies to send you >one to try. We have used them from Dako, Vector, and Biocare. We don't >use them any more. We have the coverplates from Shandon and that works >well for our hand-staining. > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 -- From rebecca.riesen <@t> dsilabs.com Wed Sep 28 16:06:59 2005 From: rebecca.riesen <@t> dsilabs.com (Riesen, Rebecca) Date: Wed Sep 28 16:07:31 2005 Subject: [Histonet] CPT FOR TOUCH PREPS Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA0AF@dsi-ex01.gateway.dom> I just attended a meeting on coding and compliance at NSH in Ft. Lauderdale and this was one of the topics. It was suggested that the code 88161 be used for touch preps, with the warning that if you use it along with a code for a frozen section it will be thrown out. You can only code for the touch prep when there was no actual frozen section performed. My understanding was that you CAN code for permanents and touch preps together, just not the frozens and the touch preps. I hope this was helpful and if I misunderstood at the meeting, please feel free to correct me. THANKS! DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From TJJ <@t> Stowers-Institute.org Wed Sep 28 16:11:12 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Sep 28 16:11:46 2005 Subject: [Histonet] Re: Methyl Green Message-ID: Chris makes some very good suggestions. We have found that tissues subjected to HIER don't want to take up the green stain using the usual staining procedures, but had not found the "magic bullet" to get them to stain satisfactorally. Other samples (no pretreatment or enzyme pretreated) counterstain very well with methyl green. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From jbaez <@t> interscopepath.com Wed Sep 28 16:13:58 2005 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Wed Sep 28 16:14:28 2005 Subject: [Histonet] cpt codes for direct smears Message-ID: <9E956D8FEB06C2408B08AC16498325E9FE43@adsl-67-113-77-28.dsl.lsan03.pacbell.net> We also bill for an 88329. Janet E. Baez Histology Manager Interscope Pathology Canoga Park, Ca. -----Original Message----- From: Maray Weirauch [mailto:mweirauch@crittenton.com] Sent: Wednesday, September 28, 2005 10:18 AM To: bwhitaker@brownpathology.com; histonet@lists.utsouthwestern.edu; Barb.Richmond@mckennan.org; jnocito@pathreflab.com Subject: RE: [Histonet] cpt codes for direct smears We have also been denied payment when using 88160 or 88161. The Centers for Medicare and Medicaid Services (CMS) have taken the stand that cytopathology codes 88160, 88161 and 88162 can not be reported for the same patient on the same date of service with codes 88304 to 88309, 'Level III through VI - Surgical pathology, gross and microscopic examination.' CMS said it considers cytopathology smears prepared from surgical pathology specimens as duplicate procedures that should not be reported separately. They will allow the use of modifier -59 for bypass if the cytopathology smears are performed on separate specimens and independent diagnoses are rendered. That scenario never seems to apply to our situation, so we also bill for a Pathology consultation during surgery, 88329. >>> "Bonnie Whitaker" 9/27/2005 10:22:49 AM >>> According to our billing company, we were being denied 88160 or 88161 if permanent sections were being done on the same specimen. We are billing an 88329 for those. What has everyone else's experience been? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 27, 2005 9:05 AM To: 'Barb Richmond'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cpt codes for direct smears We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From galinadeyneko <@t> yahoo.com Wed Sep 28 17:00:08 2005 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Wed Sep 28 17:00:27 2005 Subject: [Histonet] antibody for collagen Message-ID: <20050928220008.47992.qmail@web33106.mail.mud.yahoo.com> Dear colleagues, Would you please share information about antibodies for detecting collagen ( I do not specify type) and pre-collagen in mouse's heart and aorta FFPE tissues. I looked through the archive, but I did not find an answer. We need some additional sensitive test for collagen besides Masson Trichrome and Sirius Red stainings. Any suggestions, protocol, vendors would be highly appreciated. Thank you at advance . Galina Deyneko Novartis Cambridge MA 617-871-7613. --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From sandosis <@t> uia.net Wed Sep 28 18:00:00 2005 From: sandosis <@t> uia.net (sandosis@uia.net) Date: Wed Sep 28 18:00:28 2005 Subject: [Histonet] TS 106 Message-ID: <200509282300.j8SN02T50259@smtp2.uia.net> Hello, Is there anyone out there who is doing the TS 106 (Thymidylate Synthase)using the Ventana Benchmark? I've found the antibody at Zymed (IVD) and Chemicon (RUO) Would appreciate any info on: Source: (Zymed vs. Chemicon vs. others) Dilution: Antigen Retrieval: (CC1, CC2, mild, standard, extended) Antibody time: Thanks so much for sharing the results of your hard work. Sandra --------------------------------------------- This message was sent using the UIA Web Mail Server. ULTIMATE Internet Access, Inc http://www.uia.net/ From arvind <@t> nbrc.res.in Wed Sep 28 23:39:04 2005 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Wed Sep 28 23:37:23 2005 Subject: [Histonet] querry Message-ID: can any one pleas tell protocol for subbing slides with gelatin with some extra attachment property actually my tissue are comming off from the slides thanks Arvind Singh Pundir National Brain Rsearch Centre gurgaon, INDIA From ian.montgomery <@t> bio.gla.ac.uk Thu Sep 29 02:53:57 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Thu Sep 29 02:54:22 2005 Subject: Fwd: [Histonet] querry Message-ID: <6.2.3.4.2.20050929084427.030e3a30@udcf.gla.ac.uk> Arvind, Gelatine. = 1g. Chrome Alum. = 0.1g. (Chromic potassium sulphate) Distilled water. = 200ml. Dissolve the gelatine in the water using gentle heat then add the chrome alum. Coat the slides by dipping into the solution, draining off excess then drying in an oven. For large numbers of slides I keep clean staining racks specifically for subbing. Ian. >can any one pleas tell protocol for subbing slides with gelatin with >some extra attachment property actually my tissue are comming off >from the slides > > > >thanks > > > >Arvind Singh Pundir > >National Brain Rsearch Centre > >gurgaon, INDIA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From patrick.howorth <@t> bristol.ac.uk Thu Sep 29 04:54:23 2005 From: patrick.howorth <@t> bristol.ac.uk (PW Howorth, Physiology) Date: Thu Sep 29 05:08:29 2005 Subject: [Histonet] re: phosphate buffered saline or Tris PBS ? Message-ID: Hi, I'm currently using immunohistochemistry (free floating sections or serially mounted sections) to map the rat brain. We use viral injections to express green fluorescent protein and use 1 or 2 other labels (i.e. Cy3 or AMCA) to colocalise receptors with GFP expression. I have been trying to improve our methods that we currently use. Originally we used Tris PBS (0.01M, pH 7.4), I notice that other groups use a variety of differences in these recipes, what advantages does Tris PBS offer (if any) or would you recommend using PBS (either 0.1M or 0.01M) ? Secondly, when using triton x-100 (0.3%) has anyone noticed a reduction with GFP signal compared to just using 50% ethanol ? And finally, we use cardiac perfusion to initially fix tissue followed by 2 hours post fixation with 4% phosphate buffered (0.1M, pH 7.4) formalin. The antibody we use for mu opioid receptors (diasorin) - does anyone have any experience with this antibody and this method have you done antigen retrieval to improve the signal ? Many thanks Patrick ---------------------- Patrick Howorth, Dept of Physiology, University of Bristol, Bristol, BS8 1TD. +44 (0)117 33 17112 patrick.howorth@bristol.ac.uk From joannew <@t> bluebottle.com Thu Sep 29 07:38:22 2005 From: joannew <@t> bluebottle.com (Joanne Whitehead) Date: Thu Sep 29 07:38:42 2005 Subject: [Histonet] "empty" cryosections! Message-ID: <1127997502.433be03ea13c6@bluebottle.com> Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris From jerryd <@t> dcla.com Thu Sep 29 08:52:28 2005 From: jerryd <@t> dcla.com (Jerry Duncan) Date: Thu Sep 29 08:52:48 2005 Subject: [Histonet] gold chloride Message-ID: <6E1A8C242B4D6447A14A170D98A45C63EF18@barracuda.dcla.com> Fellow histologists, We currently prepare gold chloride rather than purchase the prepared liquid. Does gold chloride need to be refrigerated after preparation? Thanks, Jerry From rjbuesa <@t> yahoo.com Thu Sep 29 09:12:44 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Sep 29 09:13:03 2005 Subject: [Histonet] gold chloride In-Reply-To: <6E1A8C242B4D6447A14A170D98A45C63EF18@barracuda.dcla.com> Message-ID: <20050929141244.92706.qmail@web61218.mail.yahoo.com> I not only did not refrigerate it, but I used it several times until the pale yellow color began to fade. Never had any problems by doing that. Rene J. Jerry Duncan wrote: Fellow histologists, We currently prepare gold chloride rather than purchase the prepared liquid. Does gold chloride need to be refrigerated after preparation? Thanks, Jerry _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From cfavara <@t> niaid.nih.gov Thu Sep 29 09:47:07 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Thu Sep 29 09:50:42 2005 Subject: [Histonet] "empty" cryosections! Message-ID: Welcome to the world of histology - Gayle Callis is the expert to guide you. If she does not respond contact her directly! Hang in there it does get better! x Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Joanne Whitehead [mailto:joannew@bluebottle.com] Sent: Thursday, September 29, 2005 5:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "empty" cryosections! Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ian.montgomery <@t> bio.gla.ac.uk Thu Sep 29 09:59:41 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Thu Sep 29 09:59:52 2005 Subject: Fwd: [Histonet] gold chloride Message-ID: <6.2.3.4.2.20050929155750.030e4680@udcf.gla.ac.uk> Jerry, All I do is store in an amber bottle at ambient temperature. Been doing it since I was a boy with no detriment. Ian. >Fellow histologists, > >We currently prepare gold chloride rather than purchase the prepared liquid. >Does gold chloride need to be refrigerated after preparation? > >Thanks, >Jerry > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From gcallis <@t> montana.edu Thu Sep 29 10:10:55 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Sep 29 10:11:07 2005 Subject: [Histonet] querry In-Reply-To: References: Message-ID: <6.0.0.22.1.20050929090120.01b541d0@gemini.msu.montana.edu> I only use chrome gelatin for decalcified bone sections IF the plus charge (Silane coated) slides DO NOT work. You can make it up with different bloom gelatins (bloom indicating size of gelatin molecule 100 small, 275 and 300 large). The reason I do not care for gelatin, is background staining with hematoxylin, can be reduced by dipping presubbed slides in NBF 10 dips, rinsing well and storing in cool dry box. DO NOT COAT SILANE aka PLUS CHARGE SLIDES, it is an extra expense and the gelatin (protein) goes over the top of the Plus coating, rendering it useless. This is clearly stated in package inserts with Plus Charge slides. 5gm gelatin in 1 liter distilled water, add 1 gm chromium potassium sulfate, dip clean microscope glass slides in this, air dry and store in a cool dry place. You can these slides like regular slide to pick up section and do not add anything to waterbath, additional adhesives will cause more unsightly background. OR Add 10 mls of this stock subbing solution to a 2 liter waterbath, like adding commerically available adhesives to a waterbath, dry sections in normal way - We dry bone sections FLAT at 37-40C for a couple of days, overnight is too minimal. Soft tissues can be dried in an oven. Chromium is considered very toxic, so use precautions when preparing the subbing solution. We preferred the waterbath method when doing a large number of blocks. Lessens preparation time to just have it in a waterbath already. We only use this subbing solution with soft tissue as a backup when all else fails to keep sections on a slide. We never use it for immunstaining purposes. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Thu Sep 29 10:20:12 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Sep 29 10:20:28 2005 Subject: [Histonet] re: phosphate buffered saline or Tris PBS ? In-Reply-To: References: Message-ID: <6.0.0.22.1.20050929091209.01b667a8@gemini.msu.montana.edu> You can use either buffer for immunofluorescence work. We buy Dulbeccos PBS from Sigma or make up TBS. There are other PBS recipes that will work too. Our TBS is 0.05M concentratin, commonly used for IFA and IHC work. TBS (TRIS BUFFERED SALINE) STOCK 10X TBS TRIS Base (FW 121.41) 0.5M 60.57 g Sodium Chloride 89 - 90 g Dissolve in 800 ml distilled water Add 30 ml Hydrochloric Acid to adjust pH to 7.8, use magnetic stirrer with pH meter. Adjust final volume to 1 liter and store in refrigerator. If this becomes cloudy, toss and make new. Good for 1 year. 0.05M TBS WORKING BUFFER Dilute Stock 10X Buffer 1:10 100 ml Stock 10X TBS and 900 ml distilled water. Final pH can be 7.6 - 7.8, final concentration of TBS is 0.05M in 0.9% sodium chloride. TBS is a preferred buffer when you work with alkaline phosphatase immunohistochemical staining to eliminate phosphate ions that cause background staining with substrate/chromogen, but with IFA work, no real problems. GFP can be rencered nonfluoresceing by certain things, solvents are one so we never use detergents. You can find out more about GFP by going to Clontech website and accessing Living Colours Manual, free and in pfd format. Personally and if you notice a reduction in GFP signal by using certain reagents, I would eliminate them or reduce the concentration. t 03:54 AM 9/29/2005, you wrote: >Hi, > >I'm currently using immunohistochemistry (free floating sections or >serially mounted sections) to map the rat brain. We use viral injections >to express green fluorescent protein and use 1 or 2 other labels (i.e. Cy3 >or AMCA) to colocalise receptors with GFP expression. > >I have been trying to improve our methods that we currently use. >Originally we used Tris PBS (0.01M, pH 7.4), I notice that other groups >use a variety of differences in these recipes, what advantages does Tris >PBS offer (if any) or would you recommend using PBS (either 0.1M or 0.01M) ? > >Secondly, when using triton x-100 (0.3%) has anyone noticed a reduction >with GFP signal compared to just using 50% ethanol ? > >And finally, we use cardiac perfusion to initially fix tissue followed by >2 hours post fixation with 4% phosphate buffered (0.1M, pH 7.4) formalin. >The antibody we use for mu opioid receptors (diasorin) - does anyone have >any experience with this antibody and this method have you done antigen >retrieval to improve the signal ? > >Many thanks > >Patrick > >---------------------- >Patrick Howorth, >Dept of Physiology, >University of Bristol, >Bristol, >BS8 1TD. >+44 (0)117 33 17112 >patrick.howorth@bristol.ac.uk > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From SnyderW <@t> uhcwv.org Thu Sep 29 10:37:38 2005 From: SnyderW <@t> uhcwv.org (Snyder, Wendy) Date: Thu Sep 29 10:38:04 2005 Subject: [Histonet] Moh's Histotech salary range Message-ID: My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 From PMonfils <@t> Lifespan.org Thu Sep 29 10:38:30 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Sep 29 10:38:57 2005 Subject: [Histonet] querry Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175E6@lsexch.lsmaster.lifespan.org> Making up your own chrome gelatin is a bit tedious, though I did so for years before commercial products became available. Surgipath sells a chrome gelatin product called STA-ON, which I have used for several years now. A small amount of the solution added to the water bath works very well for many kinds of hard tissue, as well as necrotic tissue, blood clots, and other materials which tend to detach from the slide. It also works well on sections of polymers and other foreign substances, which the average clinical lab doesn't deal with very much, but which we have to section frequently since we are a core research facility. "Plus slides" don't work on such materials, since these materials usually do not possess the net negative charge that most tissues have. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Gayle > Callis > Sent: Thursday, September 29, 2005 8:10 AM > To: Arvind Pundir; Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] querry > > I only use chrome gelatin for decalcified bone sections IF the plus charge > > (Silane coated) slides DO NOT work. You can make it up with different > bloom gelatins (bloom indicating size of gelatin molecule 100 small, 275 > and 300 large). The reason I do not care for gelatin, is background > staining with hematoxylin, can be reduced by dipping presubbed slides in > NBF 10 dips, rinsing well and storing in cool dry box. > > DO NOT COAT SILANE aka PLUS CHARGE SLIDES, it is an extra expense and the > > gelatin (protein) goes over the top of the Plus coating, rendering it > useless. This is clearly stated in package inserts with Plus Charge > slides. > > 5gm gelatin in 1 liter distilled water, add 1 gm chromium potassium > sulfate, dip clean microscope glass slides in this, > air dry and store in a cool dry place. You can these slides like regular > slide to pick up section and do not add anything to waterbath, additional > adhesives will cause more unsightly background. > > OR > > Add 10 mls of this stock subbing solution to a 2 liter waterbath, like > adding commerically available adhesives > to a waterbath, dry sections in normal way - We dry bone sections FLAT at > 37-40C for a couple of days, overnight is too minimal. Soft tissues can > be > dried in an oven. > > Chromium is considered very toxic, so use precautions when preparing the > subbing solution. We preferred the waterbath method when doing a large > number of blocks. Lessens preparation time to just have it in a waterbath > > already. We only use this subbing solution with soft tissue as a backup > when all else fails to keep sections on a slide. We never use it for > immunstaining purposes. > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From TJasper <@t> smdc.org Thu Sep 29 11:03:54 2005 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Thu Sep 29 11:04:34 2005 Subject: FW: [Histonet] "empty" cryosections! Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA07D3907B@harrier> Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Jasper, Thomas G. Sent: Thursday, September 29, 2005 11:03 AM To: 'Favara, Cynthia (NIH/NIAID)'; ',histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] "empty" cryosections! Dear Joanne, One possible obstacle to optimal frozen tissue sectioning may be the fixation step prior to freezing. I have not used paraformaldehyde to fix and then freeze tissue for cryosectioning. However, I have been asked to cut frozen sections on tissue that has been formalin fixed. The degree of difficulty seems to increase the longer the tissue has been in a fixative. In other words, if we get it out of formalin quickly (within minutes) we'll probably get a decent section. If half the day has gone by we probably wouldn't try. Once a section is obtained the main problem is usually one of adherence. This experience is in a clinical setting, with human tissue, which leads me to my next point. Generally speaking, animal tissue is much drier, and therefore more difficult to section period. Also, if you are getting empty sections (no tissue) you may be having a problem with your supporting media (OCT, etc.). If the media is not adhering well to the tissue, it would lend itself to creating these holes. Fatty tissue is difficult to freeze as well, although I note that you have tried sectioning at different temperatures. I believe I have seen postings from others to drop the temperatures very low to help freeze the fatty tissues. I know there are others monitoring the Histonet with more knowledge about frozen sectioning. I just thought I'd share some of my experiences, hopefully it will be of some help. Good luck to you! tj Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Favara, Cynthia (NIH/NIAID) [mailto:cfavara@niaid.nih.gov] Sent: Thursday, September 29, 2005 9:47 AM To: 'Joanne Whitehead'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] "empty" cryosections! Welcome to the world of histology - Gayle Callis is the expert to guide you. If she does not respond contact her directly! Hang in there it does get better! x Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Joanne Whitehead [mailto:joannew@bluebottle.com] Sent: Thursday, September 29, 2005 5:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "empty" cryosections! Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. From Jackie.O'Connor <@t> abbott.com Thu Sep 29 11:23:33 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Sep 29 11:24:28 2005 Subject: [Histonet] Moh's Histotech salary range Message-ID: $150K per year, and I'll take the job. "Snyder, Wendy" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/29/2005 10:37 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Moh's Histotech salary range My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ian.montgomery <@t> bio.gla.ac.uk Thu Sep 29 11:25:19 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Thu Sep 29 11:26:18 2005 Subject: Fwd: RE: [Histonet] gold chloride Message-ID: <6.2.3.4.2.20050929171456.030d6d20@udcf.gla.ac.uk> Ronnie, After a lifetime in histology, the body is littered with detriment. Scotland, the usual, clear blue skies and long days of bright sunshine. The daughter has me on a pedestal and the wife thinks she won the National lottery when she got me. Yes, I'm still a sad pathetic crabbit auld bugger. Ian. >Did you ever get cured of the detriment, Ian? > >Hope everything's going well back home. > >best wishes >Ronnie > >Ronnie Houston >Director of Anatomic Pathology >Bon Secours HealthPartners Laboratories >5801 Bremo Road >Richmond, VA 23226 >(804) 2877972 >(804) 2877906 - fax >ronnie_houston@bshsi.com > > > > > >-----Original Message----- >From: Ian Montgomery [mailto:ian.montgomery@bio.gla.ac.uk] >Sent: Thursday, September 29, 2005 11:00 AM >To: histonet@lists.utsouthwestern.edu >Subject: Fwd: [Histonet] gold chloride > > >Jerry, > All I do is store in an amber bottle at ambient temperature. >Been doing it since I was a boy with no detriment. >Ian. > > > > >Fellow histologists, > > > >We currently prepare gold chloride rather than purchase the prepared >liquid. > >Does gold chloride need to be refrigerated after preparation? > > > >Thanks, > >Jerry > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Dr. Ian Montgomery, >Histotechnology, >IBLS Support Services, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07623 975451 >e-mail: ian.montgomery@bio.gla.ac.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >________________________________________________________________________________________________________________________________ >________________________________________________________________________________________________________________________________ > >The information in this communication is intended to be confidential >to the Individual(s) and/or Entity to whom it is addressed. >It may contain information of a Privileged and/or Confidential >nature, which is subject to Federal and/or State privacy regulations. >In the event that you are not the intended recipient or the agent of >the intended recipient, do not copy or use the information >contained within this communication, or allow it to be read, copied >or utilized in any manner, by any other person(s). Should >this communication be received in error, please notify the sender >immediately either by response e-mail or by phone, >and permanently delete the original e-mail, attachment(s), and any copies. Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From jqb7 <@t> cdc.gov Thu Sep 29 11:28:37 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Sep 29 11:29:02 2005 Subject: [Histonet] Moh's Histotech salary range Message-ID: Hey, I'll do it for $140,000! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Thursday, September 29, 2005 12:24 PM To: Snyder, Wendy Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Moh's Histotech salary range $150K per year, and I'll take the job. "Snyder, Wendy" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/29/2005 10:37 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Moh's Histotech salary range My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ian.montgomery <@t> bio.gla.ac.uk Thu Sep 29 11:40:55 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Thu Sep 29 11:41:15 2005 Subject: Fwd: RE: [Histonet] gold chloride Message-ID: <6.2.3.4.2.20050929172808.030e1c90@udcf.gla.ac.uk> Jennifer, Well, depending on the salt it was nerves or toning. There was me, late sixties, three piece suit with collar and tie, any mother would be proud. Sent to buy a couple of lemons for the gold staining (ok, deposition). Then the Celtic gene would kick in and I would return via a bar for a quick pint and funny thing, the chief technician always knew. Maybe it was the stupid look on my face or me making noises at the girls in the lab. Ah, girls in the lab, mmmmm, memories memories. Ian. >Ian, >What exactly were you doing with gold chloride as a boy? >have a great day. > > >---------- >From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian Montgomery >Sent: Thu 9/29/2005 9:59 AM >To: histonet@lists.utsouthwestern.edu >Subject: Fwd: [Histonet] gold chloride > >Jerry, > All I do is store in an amber bottle at ambient temperature. >Been doing it since I was a boy with no detriment. >Ian. > > > > >Fellow histologists, > > > >We currently prepare gold chloride rather than purchase the prepared liquid. > >Does gold chloride need to be refrigerated after preparation? > > > >Thanks, > >Jerry > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://l > ists.utsouthwestern.edu/mailman/listinfo/histonet > >Dr. Ian Montgomery, >Histotechnology, >IBLS Support Services, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07623 975451 >e-mail: ian.montgomery@bio.gla.ac.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From JWEEMS <@t> sjha.org Thu Sep 29 12:22:07 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Thu Sep 29 12:22:37 2005 Subject: [Histonet] cpt codes for direct smears Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01304D1B@sjhaexc02.sjha.org> It's my understanding that 88329 is a professional charge - not to be used for technical charges. Is this incorrect? Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Baez, Janet Sent: Wednesday, September 28, 2005 5:14 PM To: Maray Weirauch; bwhitaker@brownpathology.com; histonet@lists.utsouthwestern.edu; Barb.Richmond@mckennan.org; jnocito@pathreflab.com Subject: RE: [Histonet] cpt codes for direct smears We also bill for an 88329. Janet E. Baez Histology Manager Interscope Pathology Canoga Park, Ca. -----Original Message----- From: Maray Weirauch [mailto:mweirauch@crittenton.com] Sent: Wednesday, September 28, 2005 10:18 AM To: bwhitaker@brownpathology.com; histonet@lists.utsouthwestern.edu; Barb.Richmond@mckennan.org; jnocito@pathreflab.com Subject: RE: [Histonet] cpt codes for direct smears We have also been denied payment when using 88160 or 88161. The Centers for Medicare and Medicaid Services (CMS) have taken the stand that cytopathology codes 88160, 88161 and 88162 can not be reported for the same patient on the same date of service with codes 88304 to 88309, 'Level III through VI - Surgical pathology, gross and microscopic examination.' CMS said it considers cytopathology smears prepared from surgical pathology specimens as duplicate procedures that should not be reported separately. They will allow the use of modifier -59 for bypass if the cytopathology smears are performed on separate specimens and independent diagnoses are rendered. That scenario never seems to apply to our situation, so we also bill for a Pathology consultation during surgery, 88329. >>> "Bonnie Whitaker" 9/27/2005 10:22:49 AM >>> According to our billing company, we were being denied 88160 or 88161 if permanent sections were being done on the same specimen. We are billing an 88329 for those. What has everyone else's experience been? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 27, 2005 9:05 AM To: 'Barb Richmond'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cpt codes for direct smears We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Heather.A.Harper <@t> pcola.med.navy.mil Thu Sep 29 12:24:54 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Thu Sep 29 12:37:48 2005 Subject: [Histonet] MOHS Message-ID: <807FE48C5A7CC940B973B58D32E701431B13BD8B@nhpens-exch1.pcola.med.navy.mil> Message 24, I know most MOHS start at $25 minimal no experience. Those who have experience start at $30. Hope that helps. Heather A. Harper Supervisor of Histology Naval Hospital of Pensacola, FL -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 22, Issue 38 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Soft tissue digestion (Langiano, Lori, MS, HT) 2. Mayer's Haemalum reactive (Massimo) 3. RE: cpt codes for direct smears (Maray Weirauch) 4. Enzyme method for Re: [Histonet] Soft tissue digestion (Gayle Callis) 5. RE: Supervisor-Anatomic Pathology (Orr, Rebecca) 6. for anyone in pharmaceutical, contract labs, etc (Linke_Noelle) 7. Re: Pap Pen Blues (Johnson, Teri) 8. Re: Pap Pen Blues (Andrea T. Hooper) 9. CPT FOR TOUCH PREPS (Riesen, Rebecca) 10. Re: Methyl Green (Johnson, Teri) 11. RE: cpt codes for direct smears (Baez, Janet) 12. antibody for collagen (Galina Deyneko) 13. TS 106 (sandosis@uia.net) 14. querry (Arvind Pundir) 15. Fwd: [Histonet] querry (Ian Montgomery) 16. re: phosphate buffered saline or Tris PBS ? (PW Howorth, Physiology) 17. "empty" cryosections! (Joanne Whitehead) 18. gold chloride (Jerry Duncan) 19. Re: gold chloride (Rene J Buesa) 20. RE: "empty" cryosections! (Favara, Cynthia (NIH/NIAID)) 21. Fwd: [Histonet] gold chloride (Ian Montgomery) 22. Re: querry (Gayle Callis) 23. Re: re: phosphate buffered saline or Tris PBS ? (Gayle Callis) 24. Moh's Histotech salary range (Snyder, Wendy) 25. RE: querry (Monfils, Paul) 26. FW: [Histonet] "empty" cryosections! (Jasper, Thomas G.) 27. Re: Moh's Histotech salary range (Jackie M O'Connor) ---------------------------------------------------------------------- Message: 1 Date: Wed, 28 Sep 2005 10:04:52 -0700 From: "Langiano, Lori, MS, HT" Subject: [Histonet] Soft tissue digestion To: Message-ID: <32D2979CA2119041B2A84241FCD860FD1C4E43@STSM1BMSGM01> Content-Type: text/plain; charset="us-ascii" Has anyone had experience with digesting soft tissue away from an implant? I need to completely remove the tissue from an implant. Thanks in advance for any suggestions. Lori ------------------------------ Message: 2 Date: Wed, 28 Sep 2005 19:12:44 +0200 From: "Massimo" Subject: [Histonet] Mayer's Haemalum reactive To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dear Histonetters, I'd like to apologize if my question will sound too basic. I have stained a tissue with Mayer's Haemalum reactive. After I have put the specimen in water at a Ph a bit over 7, I would be expected a nuclear colour change from red "bordeaux" to blue. But it didn't happen. I fixed the specimen with Bouin's liquid and I clarified with Histoclear. I wonder why. Have you seen a behaviour of this kind? Thanks in advance for your help, Best Regards, Massimo ------------------------------ Message: 3 Date: Wed, 28 Sep 2005 13:18:02 -0400 From: "Maray Weirauch" Subject: RE: [Histonet] cpt codes for direct smears To: , , , Message-ID: Content-Type: text/plain; charset=US-ASCII We have also been denied payment when using 88160 or 88161. The Centers for Medicare and Medicaid Services (CMS) have taken the stand that cytopathology codes 88160, 88161 and 88162 can not be reported for the same patient on the same date of service with codes 88304 to 88309, 'Level III through VI - Surgical pathology, gross and microscopic examination.' CMS said it considers cytopathology smears prepared from surgical pathology specimens as duplicate procedures that should not be reported separately. They will allow the use of modifier -59 for bypass if the cytopathology smears are performed on separate specimens and independent diagnoses are rendered. That scenario never seems to apply to our situation, so we also bill for a Pathology consultation during surgery, 88329. >>> "Bonnie Whitaker" 9/27/2005 10:22:49 AM >>> According to our billing company, we were being denied 88160 or 88161 if permanent sections were being done on the same specimen. We are billing an 88329 for those. What has everyone else's experience been? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 27, 2005 9:05 AM To: 'Barb Richmond'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cpt codes for direct smears We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Wed, 28 Sep 2005 11:22:12 -0600 From: Gayle Callis Subject: Enzyme method for Re: [Histonet] Soft tissue digestion To: "Langiano, Lori, MS, HT" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050928111511.01b49278@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I used to digest soft tissue off skulls, long bones for museum teaching pieces using an enzyme detergent. Make up 10 to 20% BIZ detergent in water, heat to 80C in waterbath, and put bones, whatever in this for several hours. It will even remove fixed tissue (if you rinse it well with water before hand as NBF could inactivate the enzyme a bit. Do NOT boil. Rinse well with water afterwards, and soft tissue will release from bone, whatever. Beware, it will eat cartilage off bones. You can repeat until the implant is clean. Try to manipulate soft tissues off with gloved hands, or soft brushes. Best part of all this, is isn't smelly, and bones come out perfectly clean and white. I have a reference for this from J Microscopy, many moons ago. At 11:04 AM 9/28/2005, you wrote: >Has anyone had experience with digesting soft tissue away from an >implant? I need to completely remove the tissue from an implant. Thanks >in advance for any suggestions. >Lori >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 5 Date: Wed, 28 Sep 2005 13:13:00 -0500 From: "Orr, Rebecca" Subject: [Histonet] RE: Supervisor-Anatomic Pathology To: Message-ID: Content-Type: text/plain; charset="us-ascii" HI everyone, Anyone interested? It's a great place to work! Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- From: Wagner, Nikole Sent: Wednesday, September 28, 2005 8:29 AM To: Orr, Rebecca Subject: Supervisor-Anatomic Pathology Evanston Northwestern Healthcare is seeking a Laboratory Supervisor for our Anatomic Pathology Department at Evanston Hospital. The position requires HTL (ASCP) certification and 3-5 years of supervisory experience in a Histology Laboratory. Interested individuals can submit resumes to nwagner@enh.org or apply on-line at www.enh.org/careers. Nikole Wagner Sr. Human Resources Associate Evanston Northwestern Healthcare Phone. 847-570-1484 Fax. 847-570-1903 ------------------------------ Message: 6 Date: Wed, 28 Sep 2005 12:20:10 -0700 From: "Linke_Noelle" Subject: [Histonet] for anyone in pharmaceutical, contract labs, etc To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi All, I have a question regarding necropsy. Do you typically have one organ weigher, or does each individual do their own weighing? How many computers do you have set up in your necropsy rooms? Thank you!! Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 ------------------------------ Message: 7 Date: Wed, 28 Sep 2005 15:08:30 -0500 From: "Johnson, Teri" Subject: [Histonet] Re: Pap Pen Blues To: Message-ID: Content-Type: text/plain; charset="us-ascii" For What It's Worth (FWIW), I pretty well despise Pap pens. And I dearly LOVE Gayle's technical term (goo) for the stuff inside them. They're expensive, but see if you can get various companies to send you one to try. We have used them from Dako, Vector, and Biocare. We don't use them any more. We have the coverplates from Shandon and that works well for our hand-staining. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 Andrea, We now use Vectors ImmEdge pens - doesn't come up, and has a hint of color. Cheaper, two to a set. Can remark a slide when wet, although we do blot away moisture, but the pen goo does not lift nor flow across a section if it contacts buffer - been there and have experienced the "Pap pen oil slick syndrome" that ruins IHC when section gets goo-coated. To use ImmEdge, be sure to shake pens very vigorously to redistribute the goo (per their instructions). We vortex them hard to do that job. Maybe the pens you have now need a good vortexing - worth a try for pricey little pens. Pressing the point to start goo is still something that requires gentle tough, but I have not had too many problems with that. Flow is generally excellent, right amount. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) At 09:29 AM 9/28/2005, you wrote: >Hello All, > >I have used Zymed Mini Pap pens for years now as through trials and >tribulations and talking with coworkers we agreed it was the best out >there. The pen rarely came up or leaked and gave beautiful results. Even >withstood heat retrieval in DAKO Target Retrieval Solution with no >problems. The stuff was indestructible!! > >This is until recently ... I have noticed with the past several orders >that the formula has changed. The pap pen "ink" is now a blue green in >color (used to be more generic yellow/beige) and it does not stay on well >at all. I end up going home in a state of frustration every day as I pap >pen hundreds of slides with little success for the entirety of my >experiment. Arggggh! > >So I have two questions: > >(1) Has anyone else noticed this change in formula over the past year >or >so? (It may have been longer as I was working with a stock of pap pens >before that) >(2) Who makes your favorite pap pen? > >THANKS! >Andrea >-- > >________________________________ ------------------------------ Message: 8 Date: Wed, 28 Sep 2005 16:27:25 -0400 From: "Andrea T. Hooper" Subject: [Histonet] Re: Pap Pen Blues To: "Johnson, Teri" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Hi Teri, Thanks for the reply. Coverplates are great except for when, like in my case, you have 3-6 sections per slide each to be stained with a different antibody or dilution thereof! Thanks for the info, Andrea >For What It's Worth (FWIW), I pretty well despise Pap pens. And I >dearly LOVE Gayle's technical term (goo) for the stuff inside them. > >They're expensive, but see if you can get various companies to send you >one to try. We have used them from Dako, Vector, and Biocare. We don't >use them any more. We have the coverplates from Shandon and that works >well for our hand-staining. > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 -- ------------------------------ Message: 9 Date: Wed, 28 Sep 2005 17:06:59 -0400 From: "Riesen, Rebecca" Subject: [Histonet] CPT FOR TOUCH PREPS To: Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA0AF@dsi-ex01.gateway.dom> Content-Type: text/plain; charset="us-ascii" I just attended a meeting on coding and compliance at NSH in Ft. Lauderdale and this was one of the topics. It was suggested that the code 88161 be used for touch preps, with the warning that if you use it along with a code for a frozen section it will be thrown out. You can only code for the touch prep when there was no actual frozen section performed. My understanding was that you CAN code for permanents and touch preps together, just not the frozens and the touch preps. I hope this was helpful and if I misunderstood at the meeting, please feel free to correct me. THANKS! DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 10 Date: Wed, 28 Sep 2005 16:11:12 -0500 From: "Johnson, Teri" Subject: [Histonet] Re: Methyl Green To: Message-ID: Content-Type: text/plain; charset="us-ascii" Chris makes some very good suggestions. We have found that tissues subjected to HIER don't want to take up the green stain using the usual staining procedures, but had not found the "magic bullet" to get them to stain satisfactorally. Other samples (no pretreatment or enzyme pretreated) counterstain very well with methyl green. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 ------------------------------ Message: 11 Date: Wed, 28 Sep 2005 14:13:58 -0700 From: "Baez, Janet" Subject: RE: [Histonet] cpt codes for direct smears To: "Maray Weirauch" , , , , Message-ID: <9E956D8FEB06C2408B08AC16498325E9FE43@adsl-67-113-77-28.dsl.lsan03.pacbell.n et> Content-Type: text/plain; charset="us-ascii" We also bill for an 88329. Janet E. Baez Histology Manager Interscope Pathology Canoga Park, Ca. -----Original Message----- From: Maray Weirauch [mailto:mweirauch@crittenton.com] Sent: Wednesday, September 28, 2005 10:18 AM To: bwhitaker@brownpathology.com; histonet@lists.utsouthwestern.edu; Barb.Richmond@mckennan.org; jnocito@pathreflab.com Subject: RE: [Histonet] cpt codes for direct smears We have also been denied payment when using 88160 or 88161. The Centers for Medicare and Medicaid Services (CMS) have taken the stand that cytopathology codes 88160, 88161 and 88162 can not be reported for the same patient on the same date of service with codes 88304 to 88309, 'Level III through VI - Surgical pathology, gross and microscopic examination.' CMS said it considers cytopathology smears prepared from surgical pathology specimens as duplicate procedures that should not be reported separately. They will allow the use of modifier -59 for bypass if the cytopathology smears are performed on separate specimens and independent diagnoses are rendered. That scenario never seems to apply to our situation, so we also bill for a Pathology consultation during surgery, 88329. >>> "Bonnie Whitaker" 9/27/2005 10:22:49 AM >>> According to our billing company, we were being denied 88160 or 88161 if permanent sections were being done on the same specimen. We are billing an 88329 for those. What has everyone else's experience been? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 27, 2005 9:05 AM To: 'Barb Richmond'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cpt codes for direct smears We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Wed, 28 Sep 2005 15:00:08 -0700 (PDT) From: Galina Deyneko Subject: [Histonet] antibody for collagen To: histonet@lists.utsouthwestern.edu Message-ID: <20050928220008.47992.qmail@web33106.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Dear colleagues, Would you please share information about antibodies for detecting collagen ( I do not specify type) and pre-collagen in mouse's heart and aorta FFPE tissues. I looked through the archive, but I did not find an answer. We need some additional sensitive test for collagen besides Masson Trichrome and Sirius Red stainings. Any suggestions, protocol, vendors would be highly appreciated. Thank you at advance . Galina Deyneko Novartis Cambridge MA 617-871-7613. --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. ------------------------------ Message: 13 Date: Wed, 28 Sep 2005 18:00:00 EST From: Subject: [Histonet] TS 106 To: histonet@lists.utsouthwestern.edu Message-ID: <200509282300.j8SN02T50259@smtp2.uia.net> Hello, Is there anyone out there who is doing the TS 106 (Thymidylate Synthase)using the Ventana Benchmark? I've found the antibody at Zymed (IVD) and Chemicon (RUO) Would appreciate any info on: Source: (Zymed vs. Chemicon vs. others) Dilution: Antigen Retrieval: (CC1, CC2, mild, standard, extended) Antibody time: Thanks so much for sharing the results of your hard work. Sandra --------------------------------------------- This message was sent using the UIA Web Mail Server. ULTIMATE Internet Access, Inc http://www.uia.net/ ------------------------------ Message: 14 Date: Thu, 29 Sep 2005 10:09:04 +0530 From: "Arvind Pundir" Subject: [Histonet] querry To: Message-ID: Content-Type: text/plain; charset="utf-8" can any one pleas tell protocol for subbing slides with gelatin with some extra attachment property actually my tissue are comming off from the slides thanks Arvind Singh Pundir National Brain Rsearch Centre gurgaon, INDIA ------------------------------ Message: 15 Date: Thu, 29 Sep 2005 08:53:57 +0100 From: Ian Montgomery Subject: Fwd: [Histonet] querry To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.3.4.2.20050929084427.030e3a30@udcf.gla.ac.uk> Content-Type: text/plain; charset="us-ascii"; format=flowed Arvind, Gelatine. = 1g. Chrome Alum. = 0.1g. (Chromic potassium sulphate) Distilled water. = 200ml. Dissolve the gelatine in the water using gentle heat then add the chrome alum. Coat the slides by dipping into the solution, draining off excess then drying in an oven. For large numbers of slides I keep clean staining racks specifically for subbing. Ian. >can any one pleas tell protocol for subbing slides with gelatin with >some extra attachment property actually my tissue are comming off >from the slides > > > >thanks > > > >Arvind Singh Pundir > >National Brain Rsearch Centre > >gurgaon, INDIA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk ------------------------------ Message: 16 Date: Thu, 29 Sep 2005 10:54:23 +0100 From: "PW Howorth, Physiology" Subject: [Histonet] re: phosphate buffered saline or Tris PBS ? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Hi, I'm currently using immunohistochemistry (free floating sections or serially mounted sections) to map the rat brain. We use viral injections to express green fluorescent protein and use 1 or 2 other labels (i.e. Cy3 or AMCA) to colocalise receptors with GFP expression. I have been trying to improve our methods that we currently use. Originally we used Tris PBS (0.01M, pH 7.4), I notice that other groups use a variety of differences in these recipes, what advantages does Tris PBS offer (if any) or would you recommend using PBS (either 0.1M or 0.01M) ? Secondly, when using triton x-100 (0.3%) has anyone noticed a reduction with GFP signal compared to just using 50% ethanol ? And finally, we use cardiac perfusion to initially fix tissue followed by 2 hours post fixation with 4% phosphate buffered (0.1M, pH 7.4) formalin. The antibody we use for mu opioid receptors (diasorin) - does anyone have any experience with this antibody and this method have you done antigen retrieval to improve the signal ? Many thanks Patrick ---------------------- Patrick Howorth, Dept of Physiology, University of Bristol, Bristol, BS8 1TD. +44 (0)117 33 17112 patrick.howorth@bristol.ac.uk ------------------------------ Message: 17 Date: Thu, 29 Sep 2005 07:38:22 -0500 From: Joanne Whitehead Subject: [Histonet] "empty" cryosections! To: histonet@lists.utsouthwestern.edu Message-ID: <1127997502.433be03ea13c6@bluebottle.com> Content-Type: text/plain; charset=ISO-8859-1 Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris ------------------------------ Message: 18 Date: Thu, 29 Sep 2005 08:52:28 -0500 From: Jerry Duncan Subject: [Histonet] gold chloride To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <6E1A8C242B4D6447A14A170D98A45C63EF18@barracuda.dcla.com> Content-Type: text/plain; charset="iso-8859-1" Fellow histologists, We currently prepare gold chloride rather than purchase the prepared liquid. Does gold chloride need to be refrigerated after preparation? Thanks, Jerry ------------------------------ Message: 19 Date: Thu, 29 Sep 2005 07:12:44 -0700 (PDT) From: Rene J Buesa Subject: Re: [Histonet] gold chloride To: Jerry Duncan , "'histonet@lists.utsouthwestern.edu'" Message-ID: <20050929141244.92706.qmail@web61218.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I not only did not refrigerate it, but I used it several times until the pale yellow color began to fade. Never had any problems by doing that. Rene J. Jerry Duncan wrote: Fellow histologists, We currently prepare gold chloride rather than purchase the prepared liquid. Does gold chloride need to be refrigerated after preparation? Thanks, Jerry _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. ------------------------------ Message: 20 Date: Thu, 29 Sep 2005 10:47:07 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: RE: [Histonet] "empty" cryosections! To: 'Joanne Whitehead' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Welcome to the world of histology - Gayle Callis is the expert to guide you. If she does not respond contact her directly! Hang in there it does get better! x Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Joanne Whitehead [mailto:joannew@bluebottle.com] Sent: Thursday, September 29, 2005 5:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "empty" cryosections! Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Thu, 29 Sep 2005 15:59:41 +0100 From: Ian Montgomery Subject: Fwd: [Histonet] gold chloride To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.3.4.2.20050929155750.030e4680@udcf.gla.ac.uk> Content-Type: text/plain; charset="us-ascii"; format=flowed Jerry, All I do is store in an amber bottle at ambient temperature. Been doing it since I was a boy with no detriment. Ian. >Fellow histologists, > >We currently prepare gold chloride rather than purchase the prepared liquid. >Does gold chloride need to be refrigerated after preparation? > >Thanks, >Jerry > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk ------------------------------ Message: 22 Date: Thu, 29 Sep 2005 09:10:55 -0600 From: Gayle Callis Subject: Re: [Histonet] querry To: "Arvind Pundir" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050929090120.01b541d0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I only use chrome gelatin for decalcified bone sections IF the plus charge (Silane coated) slides DO NOT work. You can make it up with different bloom gelatins (bloom indicating size of gelatin molecule 100 small, 275 and 300 large). The reason I do not care for gelatin, is background staining with hematoxylin, can be reduced by dipping presubbed slides in NBF 10 dips, rinsing well and storing in cool dry box. DO NOT COAT SILANE aka PLUS CHARGE SLIDES, it is an extra expense and the gelatin (protein) goes over the top of the Plus coating, rendering it useless. This is clearly stated in package inserts with Plus Charge slides. 5gm gelatin in 1 liter distilled water, add 1 gm chromium potassium sulfate, dip clean microscope glass slides in this, air dry and store in a cool dry place. You can these slides like regular slide to pick up section and do not add anything to waterbath, additional adhesives will cause more unsightly background. OR Add 10 mls of this stock subbing solution to a 2 liter waterbath, like adding commerically available adhesives to a waterbath, dry sections in normal way - We dry bone sections FLAT at 37-40C for a couple of days, overnight is too minimal. Soft tissues can be dried in an oven. Chromium is considered very toxic, so use precautions when preparing the subbing solution. We preferred the waterbath method when doing a large number of blocks. Lessens preparation time to just have it in a waterbath already. We only use this subbing solution with soft tissue as a backup when all else fails to keep sections on a slide. We never use it for immunstaining purposes. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 23 Date: Thu, 29 Sep 2005 09:20:12 -0600 From: Gayle Callis Subject: Re: [Histonet] re: phosphate buffered saline or Tris PBS ? To: "PW Howorth, Physiology" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050929091209.01b667a8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed You can use either buffer for immunofluorescence work. We buy Dulbeccos PBS from Sigma or make up TBS. There are other PBS recipes that will work too. Our TBS is 0.05M concentratin, commonly used for IFA and IHC work. TBS (TRIS BUFFERED SALINE) STOCK 10X TBS TRIS Base (FW 121.41) 0.5M 60.57 g Sodium Chloride 89 - 90 g Dissolve in 800 ml distilled water Add 30 ml Hydrochloric Acid to adjust pH to 7.8, use magnetic stirrer with pH meter. Adjust final volume to 1 liter and store in refrigerator. If this becomes cloudy, toss and make new. Good for 1 year. 0.05M TBS WORKING BUFFER Dilute Stock 10X Buffer 1:10 100 ml Stock 10X TBS and 900 ml distilled water. Final pH can be 7.6 - 7.8, final concentration of TBS is 0.05M in 0.9% sodium chloride. TBS is a preferred buffer when you work with alkaline phosphatase immunohistochemical staining to eliminate phosphate ions that cause background staining with substrate/chromogen, but with IFA work, no real problems. GFP can be rencered nonfluoresceing by certain things, solvents are one so we never use detergents. You can find out more about GFP by going to Clontech website and accessing Living Colours Manual, free and in pfd format. Personally and if you notice a reduction in GFP signal by using certain reagents, I would eliminate them or reduce the concentration. t 03:54 AM 9/29/2005, you wrote: >Hi, > >I'm currently using immunohistochemistry (free floating sections or >serially mounted sections) to map the rat brain. We use viral injections >to express green fluorescent protein and use 1 or 2 other labels (i.e. Cy3 >or AMCA) to colocalise receptors with GFP expression. > >I have been trying to improve our methods that we currently use. >Originally we used Tris PBS (0.01M, pH 7.4), I notice that other groups >use a variety of differences in these recipes, what advantages does Tris >PBS offer (if any) or would you recommend using PBS (either 0.1M or 0.01M) ? > >Secondly, when using triton x-100 (0.3%) has anyone noticed a reduction >with GFP signal compared to just using 50% ethanol ? > >And finally, we use cardiac perfusion to initially fix tissue followed by >2 hours post fixation with 4% phosphate buffered (0.1M, pH 7.4) formalin. >The antibody we use for mu opioid receptors (diasorin) - does anyone have >any experience with this antibody and this method have you done antigen >retrieval to improve the signal ? > >Many thanks > >Patrick > >---------------------- >Patrick Howorth, >Dept of Physiology, >University of Bristol, >Bristol, >BS8 1TD. >+44 (0)117 33 17112 >patrick.howorth@bristol.ac.uk > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 24 Date: Thu, 29 Sep 2005 11:37:38 -0400 From: "Snyder, Wendy" Subject: [Histonet] Moh's Histotech salary range To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 ------------------------------ Message: 25 Date: Thu, 29 Sep 2005 11:38:30 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] querry To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175E6@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" Making up your own chrome gelatin is a bit tedious, though I did so for years before commercial products became available. Surgipath sells a chrome gelatin product called STA-ON, which I have used for several years now. A small amount of the solution added to the water bath works very well for many kinds of hard tissue, as well as necrotic tissue, blood clots, and other materials which tend to detach from the slide. It also works well on sections of polymers and other foreign substances, which the average clinical lab doesn't deal with very much, but which we have to section frequently since we are a core research facility. "Plus slides" don't work on such materials, since these materials usually do not possess the net negative charge that most tissues have. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Gayle > Callis > Sent: Thursday, September 29, 2005 8:10 AM > To: Arvind Pundir; Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] querry > > I only use chrome gelatin for decalcified bone sections IF the plus charge > > (Silane coated) slides DO NOT work. You can make it up with different > bloom gelatins (bloom indicating size of gelatin molecule 100 small, 275 > and 300 large). The reason I do not care for gelatin, is background > staining with hematoxylin, can be reduced by dipping presubbed slides in > NBF 10 dips, rinsing well and storing in cool dry box. > > DO NOT COAT SILANE aka PLUS CHARGE SLIDES, it is an extra expense and the > > gelatin (protein) goes over the top of the Plus coating, rendering it > useless. This is clearly stated in package inserts with Plus Charge > slides. > > 5gm gelatin in 1 liter distilled water, add 1 gm chromium potassium > sulfate, dip clean microscope glass slides in this, > air dry and store in a cool dry place. You can these slides like regular > slide to pick up section and do not add anything to waterbath, additional > adhesives will cause more unsightly background. > > OR > > Add 10 mls of this stock subbing solution to a 2 liter waterbath, like > adding commerically available adhesives > to a waterbath, dry sections in normal way - We dry bone sections FLAT at > 37-40C for a couple of days, overnight is too minimal. Soft tissues can > be > dried in an oven. > > Chromium is considered very toxic, so use precautions when preparing the > subbing solution. We preferred the waterbath method when doing a large > number of blocks. Lessens preparation time to just have it in a waterbath > > already. We only use this subbing solution with soft tissue as a backup > when all else fails to keep sections on a slide. We never use it for > immunstaining purposes. > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 26 Date: Thu, 29 Sep 2005 11:03:54 -0500 From: "Jasper, Thomas G." Subject: FW: [Histonet] "empty" cryosections! To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA07D3907B@harrier> Content-Type: text/plain; charset="iso-8859-1" Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Jasper, Thomas G. Sent: Thursday, September 29, 2005 11:03 AM To: 'Favara, Cynthia (NIH/NIAID)'; ',histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] "empty" cryosections! Dear Joanne, One possible obstacle to optimal frozen tissue sectioning may be the fixation step prior to freezing. I have not used paraformaldehyde to fix and then freeze tissue for cryosectioning. However, I have been asked to cut frozen sections on tissue that has been formalin fixed. The degree of difficulty seems to increase the longer the tissue has been in a fixative. In other words, if we get it out of formalin quickly (within minutes) we'll probably get a decent section. If half the day has gone by we probably wouldn't try. Once a section is obtained the main problem is usually one of adherence. This experience is in a clinical setting, with human tissue, which leads me to my next point. Generally speaking, animal tissue is much drier, and therefore more difficult to section period. Also, if you are getting empty sections (no tissue) you may be having a problem with your supporting media (OCT, etc.). If the media is not adhering well to the tissue, it would lend itself to creating these holes. Fatty tissue is difficult to freeze as well, although I note that you have tried sectioning at different temperatures. I believe I have seen postings from others to drop the temperatures very low to help freeze the fatty tissues. I know there are others monitoring the Histonet with more knowledge about frozen sectioning. I just thought I'd share some of my experiences, hopefully it will be of some help. Good luck to you! tj Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Favara, Cynthia (NIH/NIAID) [mailto:cfavara@niaid.nih.gov] Sent: Thursday, September 29, 2005 9:47 AM To: 'Joanne Whitehead'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] "empty" cryosections! Welcome to the world of histology - Gayle Callis is the expert to guide you. If she does not respond contact her directly! Hang in there it does get better! x Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Joanne Whitehead [mailto:joannew@bluebottle.com] Sent: Thursday, September 29, 2005 5:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "empty" cryosections! Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. ------------------------------ Message: 27 Date: Thu, 29 Sep 2005 11:23:33 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Moh's Histotech salary range To: "Snyder, Wendy" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" $150K per year, and I'll take the job. "Snyder, Wendy" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/29/2005 10:37 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Moh's Histotech salary range My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 38 **************************************** From gu.lang <@t> gmx.at Thu Sep 29 12:59:36 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Sep 29 12:59:57 2005 Subject: [Histonet] what is mohs? Message-ID: Hi, I don't know this term "mohs". Please would someone be so kind to explain. Thank you Gudrun From Melissa.Gonzalez <@t> cellgenesys.com Thu Sep 29 12:40:06 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Thu Sep 29 13:01:19 2005 Subject: [Histonet] RE: trouble with cryosections Message-ID: Dear Joan, I feel your pain. Today I am also beating my head over mouse skin sections, which have been fixed in 4% paraformaldehyde and infiltrated with sucrose. The way I freeze them has varied, as I find discrepancies which I haven't been able to solve. For one, I find that regardless of how I freeze the samples, if I cut these difficult tissues right away, they cut beautifully. However, If I have to store them in -80, regardless how long they equilibrate to -20 (we are talking hours up to 2 days) I have trouble cutting-- the tissue layers seem to split apart and the OCT doesn't seem to stick very well to the tissue (To date I have tried abt 6 brands of OCT,) regardless of the cryostat temperature. I don't understand how -80c storage would affect them so much, when they have been cryoprotected with sucrose, frozen in OCT in either an isopentane/dry ice slurry or in the cryostat (-20c), placed in sealed baggies, and put in boxes in the freezer. So why does everything (incl. any mouse organ) cut so much nicer when I remove them from sucrose solution when they are ready, embed in OCT, freeze using isopentane/dry ice, and then cut them that same day? Last week I cut these same skins fresh from freezing and embedding, no problem, and I stored them in -80. Yesterday I removed them from the -80, left them in -20 overnight, and today I want to scream and tear out my hair. This is not new to me, I have just silently grumbled all these years (some days are better than others), but I have about 300 skins to get through and I don't think I can take it! Thanks in advance for any replies!! Melissa -----Original Message----- "empty" cryosections! (Joanne Whitehead) Message: 17 Date: Thu, 29 Sep 2005 07:38:22 -0500 From: Joanne Whitehead Subject: [Histonet] "empty" cryosections! To: histonet@lists.utsouthwestern.edu Message-ID: <1127997502.433be03ea13c6@bluebottle.com> Content-Type: text/plain; charset=ISO-8859-1 Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris From pruegg <@t> ihctech.net Thu Sep 29 13:06:11 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Sep 29 13:07:09 2005 Subject: [Histonet] Moh's Histotech salary range In-Reply-To: Message-ID: <200509291806.j8TI6AAA004516@chip.viawest.net> Yea, you would have to pay me at least that (150K) to do MOHS, I find it to be very stressful and at my age I am iliminating strees in my life. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Thursday, September 29, 2005 9:24 AM To: Snyder, Wendy Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Moh's Histotech salary range $150K per year, and I'll take the job. "Snyder, Wendy" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/29/2005 10:37 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Moh's Histotech salary range My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lchung <@t> ppmh.org Thu Sep 29 13:08:23 2005 From: lchung <@t> ppmh.org (Chung, Luong) Date: Thu Sep 29 13:09:00 2005 Subject: [Histonet] MOHS Message-ID: How do you became a MOHS tech? Is there a special program for that? or MOHS(ASCP). -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Thursday, September 29, 2005 1:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] MOHS Message 24, I know most MOHS start at $25 minimal no experience. Those who have experience start at $30. Hope that helps. Heather A. Harper Supervisor of Histology Naval Hospital of Pensacola, FL -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 22, Issue 38 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Soft tissue digestion (Langiano, Lori, MS, HT) 2. Mayer's Haemalum reactive (Massimo) 3. RE: cpt codes for direct smears (Maray Weirauch) 4. Enzyme method for Re: [Histonet] Soft tissue digestion (Gayle Callis) 5. RE: Supervisor-Anatomic Pathology (Orr, Rebecca) 6. for anyone in pharmaceutical, contract labs, etc (Linke_Noelle) 7. Re: Pap Pen Blues (Johnson, Teri) 8. Re: Pap Pen Blues (Andrea T. Hooper) 9. CPT FOR TOUCH PREPS (Riesen, Rebecca) 10. Re: Methyl Green (Johnson, Teri) 11. RE: cpt codes for direct smears (Baez, Janet) 12. antibody for collagen (Galina Deyneko) 13. TS 106 (sandosis@uia.net) 14. querry (Arvind Pundir) 15. Fwd: [Histonet] querry (Ian Montgomery) 16. re: phosphate buffered saline or Tris PBS ? (PW Howorth, Physiology) 17. "empty" cryosections! (Joanne Whitehead) 18. gold chloride (Jerry Duncan) 19. Re: gold chloride (Rene J Buesa) 20. RE: "empty" cryosections! (Favara, Cynthia (NIH/NIAID)) 21. Fwd: [Histonet] gold chloride (Ian Montgomery) 22. Re: querry (Gayle Callis) 23. Re: re: phosphate buffered saline or Tris PBS ? (Gayle Callis) 24. Moh's Histotech salary range (Snyder, Wendy) 25. RE: querry (Monfils, Paul) 26. FW: [Histonet] "empty" cryosections! (Jasper, Thomas G.) 27. Re: Moh's Histotech salary range (Jackie M O'Connor) ---------------------------------------------------------------------- Message: 1 Date: Wed, 28 Sep 2005 10:04:52 -0700 From: "Langiano, Lori, MS, HT" Subject: [Histonet] Soft tissue digestion To: Message-ID: <32D2979CA2119041B2A84241FCD860FD1C4E43@STSM1BMSGM01> Content-Type: text/plain; charset="us-ascii" Has anyone had experience with digesting soft tissue away from an implant? I need to completely remove the tissue from an implant. Thanks in advance for any suggestions. Lori ------------------------------ Message: 2 Date: Wed, 28 Sep 2005 19:12:44 +0200 From: "Massimo" Subject: [Histonet] Mayer's Haemalum reactive To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dear Histonetters, I'd like to apologize if my question will sound too basic. I have stained a tissue with Mayer's Haemalum reactive. After I have put the specimen in water at a Ph a bit over 7, I would be expected a nuclear colour change from red "bordeaux" to blue. But it didn't happen. I fixed the specimen with Bouin's liquid and I clarified with Histoclear. I wonder why. Have you seen a behaviour of this kind? Thanks in advance for your help, Best Regards, Massimo ------------------------------ Message: 3 Date: Wed, 28 Sep 2005 13:18:02 -0400 From: "Maray Weirauch" Subject: RE: [Histonet] cpt codes for direct smears To: , , , Message-ID: Content-Type: text/plain; charset=US-ASCII We have also been denied payment when using 88160 or 88161. The Centers for Medicare and Medicaid Services (CMS) have taken the stand that cytopathology codes 88160, 88161 and 88162 can not be reported for the same patient on the same date of service with codes 88304 to 88309, 'Level III through VI - Surgical pathology, gross and microscopic examination.' CMS said it considers cytopathology smears prepared from surgical pathology specimens as duplicate procedures that should not be reported separately. They will allow the use of modifier -59 for bypass if the cytopathology smears are performed on separate specimens and independent diagnoses are rendered. That scenario never seems to apply to our situation, so we also bill for a Pathology consultation during surgery, 88329. >>> "Bonnie Whitaker" 9/27/2005 10:22:49 AM >>> According to our billing company, we were being denied 88160 or 88161 if permanent sections were being done on the same specimen. We are billing an 88329 for those. What has everyone else's experience been? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 27, 2005 9:05 AM To: 'Barb Richmond'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cpt codes for direct smears We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Wed, 28 Sep 2005 11:22:12 -0600 From: Gayle Callis Subject: Enzyme method for Re: [Histonet] Soft tissue digestion To: "Langiano, Lori, MS, HT" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050928111511.01b49278@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I used to digest soft tissue off skulls, long bones for museum teaching pieces using an enzyme detergent. Make up 10 to 20% BIZ detergent in water, heat to 80C in waterbath, and put bones, whatever in this for several hours. It will even remove fixed tissue (if you rinse it well with water before hand as NBF could inactivate the enzyme a bit. Do NOT boil. Rinse well with water afterwards, and soft tissue will release from bone, whatever. Beware, it will eat cartilage off bones. You can repeat until the implant is clean. Try to manipulate soft tissues off with gloved hands, or soft brushes. Best part of all this, is isn't smelly, and bones come out perfectly clean and white. I have a reference for this from J Microscopy, many moons ago. At 11:04 AM 9/28/2005, you wrote: >Has anyone had experience with digesting soft tissue away from an >implant? I need to completely remove the tissue from an implant. Thanks >in advance for any suggestions. >Lori >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 5 Date: Wed, 28 Sep 2005 13:13:00 -0500 From: "Orr, Rebecca" Subject: [Histonet] RE: Supervisor-Anatomic Pathology To: Message-ID: Content-Type: text/plain; charset="us-ascii" HI everyone, Anyone interested? It's a great place to work! Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- From: Wagner, Nikole Sent: Wednesday, September 28, 2005 8:29 AM To: Orr, Rebecca Subject: Supervisor-Anatomic Pathology Evanston Northwestern Healthcare is seeking a Laboratory Supervisor for our Anatomic Pathology Department at Evanston Hospital. The position requires HTL (ASCP) certification and 3-5 years of supervisory experience in a Histology Laboratory. Interested individuals can submit resumes to nwagner@enh.org or apply on-line at www.enh.org/careers. Nikole Wagner Sr. Human Resources Associate Evanston Northwestern Healthcare Phone. 847-570-1484 Fax. 847-570-1903 ------------------------------ Message: 6 Date: Wed, 28 Sep 2005 12:20:10 -0700 From: "Linke_Noelle" Subject: [Histonet] for anyone in pharmaceutical, contract labs, etc To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi All, I have a question regarding necropsy. Do you typically have one organ weigher, or does each individual do their own weighing? How many computers do you have set up in your necropsy rooms? Thank you!! Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 ------------------------------ Message: 7 Date: Wed, 28 Sep 2005 15:08:30 -0500 From: "Johnson, Teri" Subject: [Histonet] Re: Pap Pen Blues To: Message-ID: Content-Type: text/plain; charset="us-ascii" For What It's Worth (FWIW), I pretty well despise Pap pens. And I dearly LOVE Gayle's technical term (goo) for the stuff inside them. They're expensive, but see if you can get various companies to send you one to try. We have used them from Dako, Vector, and Biocare. We don't use them any more. We have the coverplates from Shandon and that works well for our hand-staining. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 Andrea, We now use Vectors ImmEdge pens - doesn't come up, and has a hint of color. Cheaper, two to a set. Can remark a slide when wet, although we do blot away moisture, but the pen goo does not lift nor flow across a section if it contacts buffer - been there and have experienced the "Pap pen oil slick syndrome" that ruins IHC when section gets goo-coated. To use ImmEdge, be sure to shake pens very vigorously to redistribute the goo (per their instructions). We vortex them hard to do that job. Maybe the pens you have now need a good vortexing - worth a try for pricey little pens. Pressing the point to start goo is still something that requires gentle tough, but I have not had too many problems with that. Flow is generally excellent, right amount. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) At 09:29 AM 9/28/2005, you wrote: >Hello All, > >I have used Zymed Mini Pap pens for years now as through trials and >tribulations and talking with coworkers we agreed it was the best out >there. The pen rarely came up or leaked and gave beautiful results. Even >withstood heat retrieval in DAKO Target Retrieval Solution with no >problems. The stuff was indestructible!! > >This is until recently ... I have noticed with the past several orders >that the formula has changed. The pap pen "ink" is now a blue green in >color (used to be more generic yellow/beige) and it does not stay on well >at all. I end up going home in a state of frustration every day as I pap >pen hundreds of slides with little success for the entirety of my >experiment. Arggggh! > >So I have two questions: > >(1) Has anyone else noticed this change in formula over the past year >or >so? (It may have been longer as I was working with a stock of pap pens >before that) >(2) Who makes your favorite pap pen? > >THANKS! >Andrea >-- > >________________________________ ------------------------------ Message: 8 Date: Wed, 28 Sep 2005 16:27:25 -0400 From: "Andrea T. Hooper" Subject: [Histonet] Re: Pap Pen Blues To: "Johnson, Teri" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Hi Teri, Thanks for the reply. Coverplates are great except for when, like in my case, you have 3-6 sections per slide each to be stained with a different antibody or dilution thereof! Thanks for the info, Andrea >For What It's Worth (FWIW), I pretty well despise Pap pens. And I >dearly LOVE Gayle's technical term (goo) for the stuff inside them. > >They're expensive, but see if you can get various companies to send you >one to try. We have used them from Dako, Vector, and Biocare. We don't >use them any more. We have the coverplates from Shandon and that works >well for our hand-staining. > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 -- ------------------------------ Message: 9 Date: Wed, 28 Sep 2005 17:06:59 -0400 From: "Riesen, Rebecca" Subject: [Histonet] CPT FOR TOUCH PREPS To: Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA0AF@dsi-ex01.gateway.dom> Content-Type: text/plain; charset="us-ascii" I just attended a meeting on coding and compliance at NSH in Ft. Lauderdale and this was one of the topics. It was suggested that the code 88161 be used for touch preps, with the warning that if you use it along with a code for a frozen section it will be thrown out. You can only code for the touch prep when there was no actual frozen section performed. My understanding was that you CAN code for permanents and touch preps together, just not the frozens and the touch preps. I hope this was helpful and if I misunderstood at the meeting, please feel free to correct me. THANKS! DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 10 Date: Wed, 28 Sep 2005 16:11:12 -0500 From: "Johnson, Teri" Subject: [Histonet] Re: Methyl Green To: Message-ID: Content-Type: text/plain; charset="us-ascii" Chris makes some very good suggestions. We have found that tissues subjected to HIER don't want to take up the green stain using the usual staining procedures, but had not found the "magic bullet" to get them to stain satisfactorally. Other samples (no pretreatment or enzyme pretreated) counterstain very well with methyl green. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 ------------------------------ Message: 11 Date: Wed, 28 Sep 2005 14:13:58 -0700 From: "Baez, Janet" Subject: RE: [Histonet] cpt codes for direct smears To: "Maray Weirauch" , , , , Message-ID: <9E956D8FEB06C2408B08AC16498325E9FE43@adsl-67-113-77-28.dsl.lsan03.pacbell.n et> Content-Type: text/plain; charset="us-ascii" We also bill for an 88329. Janet E. Baez Histology Manager Interscope Pathology Canoga Park, Ca. -----Original Message----- From: Maray Weirauch [mailto:mweirauch@crittenton.com] Sent: Wednesday, September 28, 2005 10:18 AM To: bwhitaker@brownpathology.com; histonet@lists.utsouthwestern.edu; Barb.Richmond@mckennan.org; jnocito@pathreflab.com Subject: RE: [Histonet] cpt codes for direct smears We have also been denied payment when using 88160 or 88161. The Centers for Medicare and Medicaid Services (CMS) have taken the stand that cytopathology codes 88160, 88161 and 88162 can not be reported for the same patient on the same date of service with codes 88304 to 88309, 'Level III through VI - Surgical pathology, gross and microscopic examination.' CMS said it considers cytopathology smears prepared from surgical pathology specimens as duplicate procedures that should not be reported separately. They will allow the use of modifier -59 for bypass if the cytopathology smears are performed on separate specimens and independent diagnoses are rendered. That scenario never seems to apply to our situation, so we also bill for a Pathology consultation during surgery, 88329. >>> "Bonnie Whitaker" 9/27/2005 10:22:49 AM >>> According to our billing company, we were being denied 88160 or 88161 if permanent sections were being done on the same specimen. We are billing an 88329 for those. What has everyone else's experience been? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 27, 2005 9:05 AM To: 'Barb Richmond'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cpt codes for direct smears We use 88160, other than gynecological for direct smears Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Monday, September 26, 2005 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cpt codes for direct smears I need to know what cpt code to use for a direct smear that is made from tissue and stained on the quick frozen section stain line. Any suggestions?? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Wed, 28 Sep 2005 15:00:08 -0700 (PDT) From: Galina Deyneko Subject: [Histonet] antibody for collagen To: histonet@lists.utsouthwestern.edu Message-ID: <20050928220008.47992.qmail@web33106.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Dear colleagues, Would you please share information about antibodies for detecting collagen ( I do not specify type) and pre-collagen in mouse's heart and aorta FFPE tissues. I looked through the archive, but I did not find an answer. We need some additional sensitive test for collagen besides Masson Trichrome and Sirius Red stainings. Any suggestions, protocol, vendors would be highly appreciated. Thank you at advance . Galina Deyneko Novartis Cambridge MA 617-871-7613. --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. ------------------------------ Message: 13 Date: Wed, 28 Sep 2005 18:00:00 EST From: Subject: [Histonet] TS 106 To: histonet@lists.utsouthwestern.edu Message-ID: <200509282300.j8SN02T50259@smtp2.uia.net> Hello, Is there anyone out there who is doing the TS 106 (Thymidylate Synthase)using the Ventana Benchmark? I've found the antibody at Zymed (IVD) and Chemicon (RUO) Would appreciate any info on: Source: (Zymed vs. Chemicon vs. others) Dilution: Antigen Retrieval: (CC1, CC2, mild, standard, extended) Antibody time: Thanks so much for sharing the results of your hard work. Sandra --------------------------------------------- This message was sent using the UIA Web Mail Server. ULTIMATE Internet Access, Inc http://www.uia.net/ ------------------------------ Message: 14 Date: Thu, 29 Sep 2005 10:09:04 +0530 From: "Arvind Pundir" Subject: [Histonet] querry To: Message-ID: Content-Type: text/plain; charset="utf-8" can any one pleas tell protocol for subbing slides with gelatin with some extra attachment property actually my tissue are comming off from the slides thanks Arvind Singh Pundir National Brain Rsearch Centre gurgaon, INDIA ------------------------------ Message: 15 Date: Thu, 29 Sep 2005 08:53:57 +0100 From: Ian Montgomery Subject: Fwd: [Histonet] querry To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.3.4.2.20050929084427.030e3a30@udcf.gla.ac.uk> Content-Type: text/plain; charset="us-ascii"; format=flowed Arvind, Gelatine. = 1g. Chrome Alum. = 0.1g. (Chromic potassium sulphate) Distilled water. = 200ml. Dissolve the gelatine in the water using gentle heat then add the chrome alum. Coat the slides by dipping into the solution, draining off excess then drying in an oven. For large numbers of slides I keep clean staining racks specifically for subbing. Ian. >can any one pleas tell protocol for subbing slides with gelatin with >some extra attachment property actually my tissue are comming off >from the slides > > > >thanks > > > >Arvind Singh Pundir > >National Brain Rsearch Centre > >gurgaon, INDIA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk ------------------------------ Message: 16 Date: Thu, 29 Sep 2005 10:54:23 +0100 From: "PW Howorth, Physiology" Subject: [Histonet] re: phosphate buffered saline or Tris PBS ? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Hi, I'm currently using immunohistochemistry (free floating sections or serially mounted sections) to map the rat brain. We use viral injections to express green fluorescent protein and use 1 or 2 other labels (i.e. Cy3 or AMCA) to colocalise receptors with GFP expression. I have been trying to improve our methods that we currently use. Originally we used Tris PBS (0.01M, pH 7.4), I notice that other groups use a variety of differences in these recipes, what advantages does Tris PBS offer (if any) or would you recommend using PBS (either 0.1M or 0.01M) ? Secondly, when using triton x-100 (0.3%) has anyone noticed a reduction with GFP signal compared to just using 50% ethanol ? And finally, we use cardiac perfusion to initially fix tissue followed by 2 hours post fixation with 4% phosphate buffered (0.1M, pH 7.4) formalin. The antibody we use for mu opioid receptors (diasorin) - does anyone have any experience with this antibody and this method have you done antigen retrieval to improve the signal ? Many thanks Patrick ---------------------- Patrick Howorth, Dept of Physiology, University of Bristol, Bristol, BS8 1TD. +44 (0)117 33 17112 patrick.howorth@bristol.ac.uk ------------------------------ Message: 17 Date: Thu, 29 Sep 2005 07:38:22 -0500 From: Joanne Whitehead Subject: [Histonet] "empty" cryosections! To: histonet@lists.utsouthwestern.edu Message-ID: <1127997502.433be03ea13c6@bluebottle.com> Content-Type: text/plain; charset=ISO-8859-1 Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris ------------------------------ Message: 18 Date: Thu, 29 Sep 2005 08:52:28 -0500 From: Jerry Duncan Subject: [Histonet] gold chloride To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <6E1A8C242B4D6447A14A170D98A45C63EF18@barracuda.dcla.com> Content-Type: text/plain; charset="iso-8859-1" Fellow histologists, We currently prepare gold chloride rather than purchase the prepared liquid. Does gold chloride need to be refrigerated after preparation? Thanks, Jerry ------------------------------ Message: 19 Date: Thu, 29 Sep 2005 07:12:44 -0700 (PDT) From: Rene J Buesa Subject: Re: [Histonet] gold chloride To: Jerry Duncan , "'histonet@lists.utsouthwestern.edu'" Message-ID: <20050929141244.92706.qmail@web61218.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I not only did not refrigerate it, but I used it several times until the pale yellow color began to fade. Never had any problems by doing that. Rene J. Jerry Duncan wrote: Fellow histologists, We currently prepare gold chloride rather than purchase the prepared liquid. Does gold chloride need to be refrigerated after preparation? Thanks, Jerry _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. ------------------------------ Message: 20 Date: Thu, 29 Sep 2005 10:47:07 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: RE: [Histonet] "empty" cryosections! To: 'Joanne Whitehead' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Welcome to the world of histology - Gayle Callis is the expert to guide you. If she does not respond contact her directly! Hang in there it does get better! x Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Joanne Whitehead [mailto:joannew@bluebottle.com] Sent: Thursday, September 29, 2005 5:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "empty" cryosections! Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Thu, 29 Sep 2005 15:59:41 +0100 From: Ian Montgomery Subject: Fwd: [Histonet] gold chloride To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.3.4.2.20050929155750.030e4680@udcf.gla.ac.uk> Content-Type: text/plain; charset="us-ascii"; format=flowed Jerry, All I do is store in an amber bottle at ambient temperature. Been doing it since I was a boy with no detriment. Ian. >Fellow histologists, > >We currently prepare gold chloride rather than purchase the prepared liquid. >Does gold chloride need to be refrigerated after preparation? > >Thanks, >Jerry > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk ------------------------------ Message: 22 Date: Thu, 29 Sep 2005 09:10:55 -0600 From: Gayle Callis Subject: Re: [Histonet] querry To: "Arvind Pundir" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050929090120.01b541d0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I only use chrome gelatin for decalcified bone sections IF the plus charge (Silane coated) slides DO NOT work. You can make it up with different bloom gelatins (bloom indicating size of gelatin molecule 100 small, 275 and 300 large). The reason I do not care for gelatin, is background staining with hematoxylin, can be reduced by dipping presubbed slides in NBF 10 dips, rinsing well and storing in cool dry box. DO NOT COAT SILANE aka PLUS CHARGE SLIDES, it is an extra expense and the gelatin (protein) goes over the top of the Plus coating, rendering it useless. This is clearly stated in package inserts with Plus Charge slides. 5gm gelatin in 1 liter distilled water, add 1 gm chromium potassium sulfate, dip clean microscope glass slides in this, air dry and store in a cool dry place. You can these slides like regular slide to pick up section and do not add anything to waterbath, additional adhesives will cause more unsightly background. OR Add 10 mls of this stock subbing solution to a 2 liter waterbath, like adding commerically available adhesives to a waterbath, dry sections in normal way - We dry bone sections FLAT at 37-40C for a couple of days, overnight is too minimal. Soft tissues can be dried in an oven. Chromium is considered very toxic, so use precautions when preparing the subbing solution. We preferred the waterbath method when doing a large number of blocks. Lessens preparation time to just have it in a waterbath already. We only use this subbing solution with soft tissue as a backup when all else fails to keep sections on a slide. We never use it for immunstaining purposes. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 23 Date: Thu, 29 Sep 2005 09:20:12 -0600 From: Gayle Callis Subject: Re: [Histonet] re: phosphate buffered saline or Tris PBS ? To: "PW Howorth, Physiology" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050929091209.01b667a8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed You can use either buffer for immunofluorescence work. We buy Dulbeccos PBS from Sigma or make up TBS. There are other PBS recipes that will work too. Our TBS is 0.05M concentratin, commonly used for IFA and IHC work. TBS (TRIS BUFFERED SALINE) STOCK 10X TBS TRIS Base (FW 121.41) 0.5M 60.57 g Sodium Chloride 89 - 90 g Dissolve in 800 ml distilled water Add 30 ml Hydrochloric Acid to adjust pH to 7.8, use magnetic stirrer with pH meter. Adjust final volume to 1 liter and store in refrigerator. If this becomes cloudy, toss and make new. Good for 1 year. 0.05M TBS WORKING BUFFER Dilute Stock 10X Buffer 1:10 100 ml Stock 10X TBS and 900 ml distilled water. Final pH can be 7.6 - 7.8, final concentration of TBS is 0.05M in 0.9% sodium chloride. TBS is a preferred buffer when you work with alkaline phosphatase immunohistochemical staining to eliminate phosphate ions that cause background staining with substrate/chromogen, but with IFA work, no real problems. GFP can be rencered nonfluoresceing by certain things, solvents are one so we never use detergents. You can find out more about GFP by going to Clontech website and accessing Living Colours Manual, free and in pfd format. Personally and if you notice a reduction in GFP signal by using certain reagents, I would eliminate them or reduce the concentration. t 03:54 AM 9/29/2005, you wrote: >Hi, > >I'm currently using immunohistochemistry (free floating sections or >serially mounted sections) to map the rat brain. We use viral injections >to express green fluorescent protein and use 1 or 2 other labels (i.e. Cy3 >or AMCA) to colocalise receptors with GFP expression. > >I have been trying to improve our methods that we currently use. >Originally we used Tris PBS (0.01M, pH 7.4), I notice that other groups >use a variety of differences in these recipes, what advantages does Tris >PBS offer (if any) or would you recommend using PBS (either 0.1M or 0.01M) ? > >Secondly, when using triton x-100 (0.3%) has anyone noticed a reduction >with GFP signal compared to just using 50% ethanol ? > >And finally, we use cardiac perfusion to initially fix tissue followed by >2 hours post fixation with 4% phosphate buffered (0.1M, pH 7.4) formalin. >The antibody we use for mu opioid receptors (diasorin) - does anyone have >any experience with this antibody and this method have you done antigen >retrieval to improve the signal ? > >Many thanks > >Patrick > >---------------------- >Patrick Howorth, >Dept of Physiology, >University of Bristol, >Bristol, >BS8 1TD. >+44 (0)117 33 17112 >patrick.howorth@bristol.ac.uk > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 24 Date: Thu, 29 Sep 2005 11:37:38 -0400 From: "Snyder, Wendy" Subject: [Histonet] Moh's Histotech salary range To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 ------------------------------ Message: 25 Date: Thu, 29 Sep 2005 11:38:30 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] querry To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175E6@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" Making up your own chrome gelatin is a bit tedious, though I did so for years before commercial products became available. Surgipath sells a chrome gelatin product called STA-ON, which I have used for several years now. A small amount of the solution added to the water bath works very well for many kinds of hard tissue, as well as necrotic tissue, blood clots, and other materials which tend to detach from the slide. It also works well on sections of polymers and other foreign substances, which the average clinical lab doesn't deal with very much, but which we have to section frequently since we are a core research facility. "Plus slides" don't work on such materials, since these materials usually do not possess the net negative charge that most tissues have. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Gayle > Callis > Sent: Thursday, September 29, 2005 8:10 AM > To: Arvind Pundir; Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] querry > > I only use chrome gelatin for decalcified bone sections IF the plus charge > > (Silane coated) slides DO NOT work. You can make it up with different > bloom gelatins (bloom indicating size of gelatin molecule 100 small, 275 > and 300 large). The reason I do not care for gelatin, is background > staining with hematoxylin, can be reduced by dipping presubbed slides in > NBF 10 dips, rinsing well and storing in cool dry box. > > DO NOT COAT SILANE aka PLUS CHARGE SLIDES, it is an extra expense and the > > gelatin (protein) goes over the top of the Plus coating, rendering it > useless. This is clearly stated in package inserts with Plus Charge > slides. > > 5gm gelatin in 1 liter distilled water, add 1 gm chromium potassium > sulfate, dip clean microscope glass slides in this, > air dry and store in a cool dry place. You can these slides like regular > slide to pick up section and do not add anything to waterbath, additional > adhesives will cause more unsightly background. > > OR > > Add 10 mls of this stock subbing solution to a 2 liter waterbath, like > adding commerically available adhesives > to a waterbath, dry sections in normal way - We dry bone sections FLAT at > 37-40C for a couple of days, overnight is too minimal. Soft tissues can > be > dried in an oven. > > Chromium is considered very toxic, so use precautions when preparing the > subbing solution. We preferred the waterbath method when doing a large > number of blocks. Lessens preparation time to just have it in a waterbath > > already. We only use this subbing solution with soft tissue as a backup > when all else fails to keep sections on a slide. We never use it for > immunstaining purposes. > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 26 Date: Thu, 29 Sep 2005 11:03:54 -0500 From: "Jasper, Thomas G." Subject: FW: [Histonet] "empty" cryosections! To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA07D3907B@harrier> Content-Type: text/plain; charset="iso-8859-1" Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Jasper, Thomas G. Sent: Thursday, September 29, 2005 11:03 AM To: 'Favara, Cynthia (NIH/NIAID)'; ',histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] "empty" cryosections! Dear Joanne, One possible obstacle to optimal frozen tissue sectioning may be the fixation step prior to freezing. I have not used paraformaldehyde to fix and then freeze tissue for cryosectioning. However, I have been asked to cut frozen sections on tissue that has been formalin fixed. The degree of difficulty seems to increase the longer the tissue has been in a fixative. In other words, if we get it out of formalin quickly (within minutes) we'll probably get a decent section. If half the day has gone by we probably wouldn't try. Once a section is obtained the main problem is usually one of adherence. This experience is in a clinical setting, with human tissue, which leads me to my next point. Generally speaking, animal tissue is much drier, and therefore more difficult to section period. Also, if you are getting empty sections (no tissue) you may be having a problem with your supporting media (OCT, etc.). If the media is not adhering well to the tissue, it would lend itself to creating these holes. Fatty tissue is difficult to freeze as well, although I note that you have tried sectioning at different temperatures. I believe I have seen postings from others to drop the temperatures very low to help freeze the fatty tissues. I know there are others monitoring the Histonet with more knowledge about frozen sectioning. I just thought I'd share some of my experiences, hopefully it will be of some help. Good luck to you! tj Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Favara, Cynthia (NIH/NIAID) [mailto:cfavara@niaid.nih.gov] Sent: Thursday, September 29, 2005 9:47 AM To: 'Joanne Whitehead'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] "empty" cryosections! Welcome to the world of histology - Gayle Callis is the expert to guide you. If she does not respond contact her directly! Hang in there it does get better! x Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Joanne Whitehead [mailto:joannew@bluebottle.com] Sent: Thursday, September 29, 2005 5:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "empty" cryosections! Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. ------------------------------ Message: 27 Date: Thu, 29 Sep 2005 11:23:33 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Moh's Histotech salary range To: "Snyder, Wendy" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" $150K per year, and I'll take the job. "Snyder, Wendy" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/29/2005 10:37 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Moh's Histotech salary range My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 38 **************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Thu Sep 29 13:27:52 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Thu Sep 29 13:28:47 2005 Subject: [Histonet] RE: trouble with cryosections Message-ID: <5784D843593D874C93E9BADCB87342AB9166AB@tpiserver03.Coretech-holdings.com> Tissue must be snap frozen (within a second or two) to avoid crystal ice formation in the cells. Ice in vitreous form is amorphous, and does not expand on freezing. Expansion on freezing is a unique property of ice crystals. However, vitreous ice is an unstable state of nature. Even stored below -80, it will gradually form as crystals, and expand. The expansion tears cell membranes, leaks contents, and generally messes up the tissue for histology, probably also including the cutting properties. See the link below for a full discussion http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Freez ing%20Artifact.pdf Absolutely, cut as soon as possible after freezing as possible, within hours, not days. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Melissa Gonzalez Sent: Thursday, September 29, 2005 12:40 PM To: histonet@lists.utsouthwestern.edu Cc: joannew@bluebottle.com Subject: [Histonet] RE: trouble with cryosections Dear Joan, I feel your pain. Today I am also beating my head over mouse skin sections, which have been fixed in 4% paraformaldehyde and infiltrated with sucrose. The way I freeze them has varied, as I find discrepancies which I haven't been able to solve. For one, I find that regardless of how I freeze the samples, if I cut these difficult tissues right away, they cut beautifully. However, If I have to store them in -80, regardless how long they equilibrate to -20 (we are talking hours up to 2 days) I have trouble cutting-- the tissue layers seem to split apart and the OCT doesn't seem to stick very well to the tissue (To date I have tried abt 6 brands of OCT,) regardless of the cryostat temperature. I don't understand how -80c storage would affect them so much, when they have been cryoprotected with sucrose, frozen in OCT in either an isopentane/dry ice slurry or in the cryostat (-20c), placed in sealed baggies, and put in boxes in the freezer. So why does everything (incl. any mouse organ) cut so much nicer when I remove them from sucrose solution when they are ready, embed in OCT, freeze using isopentane/dry ice, and then cut them that same day? Last week I cut these same skins fresh from freezing and embedding, no problem, and I stored them in -80. Yesterday I removed them from the -80, left them in -20 overnight, and today I want to scream and tear out my hair. This is not new to me, I have just silently grumbled all these years (some days are better than others), but I have about 300 skins to get through and I don't think I can take it! Thanks in advance for any replies!! Melissa -----Original Message----- "empty" cryosections! (Joanne Whitehead) Message: 17 Date: Thu, 29 Sep 2005 07:38:22 -0500 From: Joanne Whitehead Subject: [Histonet] "empty" cryosections! To: histonet@lists.utsouthwestern.edu Message-ID: <1127997502.433be03ea13c6@bluebottle.com> Content-Type: text/plain; charset=ISO-8859-1 Hi everyone, I'm relatively new at cryosectioning, and so far have found it to be a very frustrating experience! Some days it goes very smoothly and I get plenty of nice sections, but most often I feel like it's a battle of wills with the cryostat, which ususally wins. My major problem is getting "empty" sections - the media cuts smoothly but the tissue itself curls or crumples up, leaving just an open circle of media. The other difficulty is with sections wrinkling as they come off the knife, instead of lying flat. Does anyone know why this happens, or how I can prevent it? I'm sectioning mouse intestinal tissue, which has been rolled into a "Swiss roll", fixed in paraformaldehyde, infused with sucrose, snap frozen in isopentane over liquid nitrogen and embedded in OTC. I have tried cutting at different temperatures, from -18C to -30C, with my samples equilibrating in the machine at least an hour before I start. I have seen protocols in which the tissue is embedded in OTC before freezing, and also infusing the tissue with a mixture of sucrose and OTC before embedding. Does anyone have any preference for these methods? I would really appreciate any advice! Thank you! Joanne Curie Institute, Paris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Thu Sep 29 13:31:29 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Sep 29 13:32:24 2005 Subject: [Histonet] Moh's Histotech salary range Message-ID: Hello, I live in Portland, Or and the salary here is about 36-39 grand a year, but I am not ASCP certified. I make about $3-4 dollars less than a certifed tech. Hope that helps. Robyn OHSU >>> "Snyder, Wendy" 9/29/2005 8:37 AM >>> My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Thu Sep 29 13:54:52 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Sep 29 13:55:22 2005 Subject: [Histonet] what is mohs? Message-ID: It is particular technique that was invented by a man named Frederick Mohs. The technique is that a surgeon is technically the pathologist reading his/her own slides while the patient waits, whereas normally a pathologist would read the slide. And the other unique thing about this particular surgery is that you cut the tissue tangential and not breadloafed. Robyn OHSU >>> "Gudrun Lang" 9/29/2005 10:59 AM >>> Hi, I don't know this term "mohs". Please would someone be so kind to explain. Thank you Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From haldana <@t> unimoron.edu.ar Thu Sep 29 14:03:00 2005 From: haldana <@t> unimoron.edu.ar (Hernan Aldana Marcos) Date: Thu Sep 29 14:05:07 2005 Subject: [Histonet] brain tissue Message-ID: <004301c5c528$69727680$a904a8c0@um.edu> Dear Histonetters I work with rat?s brains fixed in paraformaldehyde. I have problems with paraffin inclusion and section. I always obtain brittle sections. Help me Thanks in advance Dr. Hern?n Aldana Marcos haldana@unimoron.edu.ar From jqb7 <@t> cdc.gov Thu Sep 29 14:05:29 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Sep 29 14:06:17 2005 Subject: [Histonet] what is mohs? Message-ID: Found this on-line: Mohs micrographic surgery for nonmelanoma skin cancer Surgery Overview Mohs micrographic surgery involves removing a skin cancer one layer at a time and examining these layers under a microscope immediately after they are removed. This procedure allows for a close examination of each layer of skin to detect cancer cells. It also allows a minimal amount of tissue to be removed while ensuring complete removal of all the cancer cells. A local anesthetic is injected into the skin before the surgery. Your doctor then begins to remove the skin cancer and a small amount of healthy tissue, one layer of skin at a time. Each tissue layer is prepared and examined under the microscope for cancer cells. Surgery is complete when no more cancer cells are detected -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Thursday, September 29, 2005 2:00 PM To: Histonetliste (Histonetliste) Subject: [Histonet] what is mohs? Hi, I don't know this term "mohs". Please would someone be so kind to explain. Thank you Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mgr <@t> seattleskincancer.com Thu Sep 29 14:35:39 2005 From: mgr <@t> seattleskincancer.com (Manager) Date: Thu Sep 29 14:41:47 2005 Subject: [Histonet] what is mohs? In-Reply-To: Message-ID: <20050929194126.3E1EC6FE5B@smtp3.pacifier.net> The Mohs technique is used primarily for skin cancer, and done using frozen sections. There is a society called the American Society for Mohs Histotechnology. No certification at this time specifically for Mohs, although it is always being talked about. Beth Seattle Skin Cancer Center -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Thursday, September 29, 2005 11:55 AM To: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] what is mohs? It is particular technique that was invented by a man named Frederick Mohs. The technique is that a surgeon is technically the pathologist reading his/her own slides while the patient waits, whereas normally a pathologist would read the slide. And the other unique thing about this particular surgery is that you cut the tissue tangential and not breadloafed. Robyn OHSU >>> "Gudrun Lang" 9/29/2005 10:59 AM >>> Hi, I don't know this term "mohs". Please would someone be so kind to explain. Thank you Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rdavis4 <@t> rdg.boehringer-ingelheim.com Thu Sep 29 14:41:09 2005 From: rdavis4 <@t> rdg.boehringer-ingelheim.com (rdavis4@rdg.boehringer-ingelheim.com) Date: Thu Sep 29 14:42:07 2005 Subject: [Histonet] brain tissue Message-ID: Try fixing the brains in 10% NBF instead. We do rat brains all the time, and the sections are beautiful. Also, check your dehydration times in your processing schedule. Rebecca A. Davis, A.A.S., NYS LVT, HT (ASCP) Toxicology, Histopathology Lab Boehringer-Ingelheim Pharmaceuticals, Inc. rdavis4@rdg.boehringer-ingelheim.com 203-798-5448 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Thursday, September 29, 2005 3:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] brain tissue Dear Histonetters I work with rat?s brains fixed in paraformaldehyde. I have problems with paraffin inclusion and section. I always obtain brittle sections. Help me Thanks in advance Dr. Hern?n Aldana Marcos haldana@unimoron.edu.ar _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.ingles <@t> hosp.wisc.edu Thu Sep 29 14:54:09 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Thu Sep 29 14:54:28 2005 Subject: [Histonet] Moh's Histotech salary range Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE5A@uwhis-xchng2.hosp.wisc.edu> Gee, I guess I have to talk to my boss about a raise... I work in Madison, Wisc (Home of Dr. Mohs) and I only get $19.45/hr. I have am lead tech, have an HTL, and 4 years experience with 1 of those in Mohs. Darn these big hospitals. :) I have actually gotten a total of about $3/hr. raise this year though. Go unions! Claire Ingles UW West MOHS Clinic Madison WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Robyn Vazquez Sent: Thu 9/29/2005 1:31 PM To: histonet@lists.utsouthwestern.edu; SnyderW@uhcwv.org Cc: Subject: Re: [Histonet] Moh's Histotech salary range Hello, I live in Portland, Or and the salary here is about 36-39 grand a year, but I am not ASCP certified. I make about $3-4 dollars less than a certifed tech. Hope that helps. Robyn OHSU >>> "Snyder, Wendy" 9/29/2005 8:37 AM >>> My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Thu Sep 29 15:22:41 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Thu Sep 29 15:23:04 2005 Subject: [Histonet] IHC questions Message-ID: <20050929202241.58923.qmail@web90210.mail.scd.yahoo.com> I'm staining rat liver for anti-Brdu next week, AEC with crystal mount. I'm assuming the aquesous Hematoxylin mentioned in my "old" procedure referes to Mayers. Is that correct? Is 5 minutes sufficient and can scotts be used for bluing or should I use a tap rinse? I know fixation is routinely 24 hours. To assure complete fixation would 48 hours work ok. I need to do AR anyways. Thanks for everyones answers. Been doing this for over 13 years but only routine. This IHC, IF, and research stuff is a "tad" new. Thanks again, Steve t"T"t --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From cwscouten <@t> myneurolab.com Thu Sep 29 15:31:35 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Thu Sep 29 15:32:23 2005 Subject: [Histonet] brain tissue Message-ID: <5784D843593D874C93E9BADCB87342AB9166B1@tpiserver03.Coretech-holdings.com> May I ask, are the brains perfused or immersed? Perfusion gives better tissue quality, since the whole brain is a big chunck for immersion. Interior poorly fixed is one interpretation of the brittle problem. I have not used paraffin, can cannot speak to the inclusion problem. Why not freeze the brain and cut whole sections? Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hernan Aldana Marcos Sent: Thursday, September 29, 2005 2:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] brain tissue Dear Histonetters I work with rat?s brains fixed in paraformaldehyde. I have problems with paraffin inclusion and section. I always obtain brittle sections. Help me Thanks in advance Dr. Hern?n Aldana Marcos haldana@unimoron.edu.ar _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Sep 29 16:27:34 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Sep 29 16:28:09 2005 Subject: [Histonet] Hotel key cards, potential identity theft during travel - sent to our university faculty Message-ID: <6.0.0.22.1.20050929151625.01b3d0a8@gemini.msu.montana.edu> Dear All This message was sent to my chemistry/faculty hubby (should have included staff also!!) via accounting technician in his department at our university. I thought it interesting, particularly since recent NSH convention attendance in Ft. Lauderdale. So this is just a bit of heads up alert during your travels to protect your credit, etc, etc. You can pass it on to others without all the names included. Gayle Callis Veterinary Molecular Biology Montana State University - Bozeman Bozeman MT 59717-3610 >Date: Thu, 29 Sep 2005 13:11:10 -0600 (MDT) >Subject: Hotel Key Cards - Potential Identity Theft > >I was at a recent purchasing card meeting and the following information >was one of the topics. Thought everyone would be interested in this info >as we usually all travel and stay in motels at various times. I hadn't >even thought of this - just another was for potential identity theft! > >This info was verified by the Purchasing Dept as having been a topic at a >regional meeting with the information being presented in a Identity Theft >Presentation by a police department. > >The following information is embedded in the credit card type hotel room >keys used throughout the industry: > >Customers name >Customers partial home address >Hotel room number >Check in date and check out date >AND >your credit card number and expiration date!! > >When you turn in the key to the front desk - this information is available >for any employee to scan and access or take home and use a scanning >device, access the information onto a laptop computer and go shopping at >your expense. Your information is overwritten only when a new guest is >assigned the card. > >The bottom line is - Keep the cards - take them home with you or destroy >them. NEVER leave them behind in the room or room wastebasket, and NEVER >turn them in to the front desk when you check out of a room. They will >not charge your for the card (it's illegal) and you won't be leaving >behind valuable personal information. > >Make sure you destroy the card and cut through the electonic information >strip - don't just toss in the trash can. > >S White >Accounting Technician >Montana State University >Department of Chemistry/Biochemistry >Bozeman, MT 59717 From kgibbon <@t> qltinc.com Thu Sep 29 16:49:10 2005 From: kgibbon <@t> qltinc.com (kgibbon@qltinc.com) Date: Thu Sep 29 16:49:34 2005 Subject: [Histonet] Cryostat decontamination Message-ID: Thanks Jan for the interesting message on disinfection. I have usually used Formalin in the past on all the old cryostats but I remember buying a solution from Leica called "cryofect" which (I think) is chlorhexidine in 70% isopropanol. Is it still marketed by Leica? as I remember it was to be used without defrosting the cryostat. Kevin Gibbon QLT Inc. |---------+-------------------------------------------> | | | | | Sent by: | | | | | | | | | | | | 09/26/2005 03:14 PM | | | | |---------+-------------------------------------------> >------------------------------------------------------------------------------------------------------------------------------| | | | To: | | cc: , | | Subject: [Histonet] Cryostat decontamination | >------------------------------------------------------------------------------------------------------------------------------| Hi Dorothy and Histonet subscribers, I'm writing in response to comments I've seen in the past few days about UV disinfection procedures for cryostats. I'm not sure if the listing on the spreadsheet from Gloria's workshop referred to our instrument or not, but Leica recently released the CM1850 UV cryostat with ultraviolet light (UVC) disinfection...so I thought I should chime in. We had the same questions about cryostat disinfection that many of you have had. Therefore, as part of our development process we hired an independent laboratory to perform tests that would verify the efficacy of UVC exposure against specific pathogens in the CM1850 at cold temperatures. The results assured us that our system is a convenient and safe means of inactivating microorganisms in the air and on exposed surfaces at temperatures down to -20?C and that using the system significantly reduces the risk of infection to the operator. We proudly provide a CD that contains the certificate from the consultant, details of how the tests were performed and the results that were obtained. Whether a cryostat has a built-in disinfection system of any kind or not, there are several very important things to remember about disinfecting cryostats. 1. Before beginning a disinfection protocol, don personal protective equipment (puncture and penetration resistant gloves, gowns, etc). 2. Remove all debris from the cryostat and disposed of it according to the policies and procedures of your institution. The debris must be removed because organic material (blood and proteins) may contain high concentrations of microorganisms and could possibly inactivate chemical germicides or prevent access to contaminated surfaces. 3. Use 70% ethyl alcohol to clean the cryostat. The germicidal activity of ethyl alcohol is most effective in that range and it has an advantage over isopropyl alcohol of being able to kill hydrophilic viruses. 4. For those of us in the USA (other countries have access to other products), the EPA maintains a list of Antimicrobial Chemical/Registration Number Indexes and it is posted on their website http://www.epa.gov/oppad001/chemregindex.htm. From this link you can find agents effective against bloodborne pathogens such as Mycobacterium tuberculosis, human HIV-1 virus, Hepatitis B or Hepatitis C virus. It is critical to remember that NONE of these solutions have been tested at low temperatures and they can only be used at room temperature. 5. Do not create aerosols by spraying disinfectant (or anything else) in an open cryostat chamber. Pour disinfectants onto absorbent disposable towels and allow them to remain in contact with contaminated surfaces for the length of time specified in the instructions of the individual agents. I hope this information is useful. Please let me know if you have any questions. Best wishes to all, Jan Minshew HT, HTL(ASCP) Marketing Manager Leica Microsystems, Inc. 2345 Waukegan Rd Bannockburn, IL 60015 800.248.0123 x7051 Traczyk7@aol.com Sent by: To: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: (bcc: Jan Minshew/USDER/West/Leica) western.edu Subject: Re: [Histonet] cryostat decontamination 09/26/2005 09:59 AM Greetings, I recently attended Gloria Limetti's seminar at NSH about cryostat and microtome features. She is from the University of Pitssburgh. Attendees were given an extensive spreadsheet listing which features were available on which models. In an effort not to come across as non vendor specific, Gloria assigned all the companies letters. Just looking over the features, it's fairly easy to decipher which company is which. I don't have my copy in front of me now but I believe there are 7 models with various types of decontamination systems offered. Hacker Instruments SL5000 is one of the units that is available with an automatic decontamination feature, perhaps Gloria will post the names of the other companies. As for UV decontamination, it is my understanding the the UV light is only effective on the surfaces of the cryostat and microtome chamber where the light actually comes into contact. Nooks and cranies would still need to be wiped down manually. As with Vinnie, I would be interested in reading a study or two on it's effectiveness in histology applications. Regards, Dorothy Murphy Traczyk National Sales Manager Hacker Instruments & Industries Inc. PO Box 1176 Winnsboro, SC 29180 1-800-442-2537 hackerlab@aol.com _www.hacker_ (http://www.hacker/) insruments.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This electronic transmission (including any and all attachments) is intended solely for the use of the individual or entity to whom it is addressed and may contain information that is privileged and/or confidential. If you are not the intended recipient of this electronic transmission, you are hereby notified that any disclosure, copying or distribution, or the taking of any action in reliance upon the contents of this electronic transmission, is strictly prohibited, and you are further requested to purge this electronic transmission and all copies thereof from your computer system. From gcallis <@t> montana.edu Thu Sep 29 17:00:25 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Sep 29 17:00:57 2005 Subject: [Histonet] Hotel key card/credit theft could be a rumor too Message-ID: <6.0.0.22.1.20050929155459.01b78480@gemini.msu.montana.edu> Apparently the hotel key card could be rumor too. You can choose for yourself what you want to do with keys by going to this link, to read up on what hotels may or may not do. Pass it on to others, as before. I'm not in panic mode, but I may bring the card with me in future, since they are free and I like cutting up things. >http://www.truthorfiction.com/rumors/k/keycards.htm Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From vazquezr <@t> ohsu.edu Thu Sep 29 17:21:54 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Sep 29 17:22:32 2005 Subject: [Histonet] Moh's Histotech salary range Message-ID: I was taught on the job about 5 years ago, but I have done histology since 1989 in the military and a private lab. Mohs was new to me when I started here. Robyn OHSU >>> "Robyn Vazquez" 9/29/2005 11:31 AM >>> Hello, I live in Portland, Or and the salary here is about 36-39 grand a year, but I am not ASCP certified. I make about $3-4 dollars less than a certifed tech. Hope that helps. Robyn OHSU >>> "Snyder, Wendy" 9/29/2005 8:37 AM >>> My hospital is in the process of starting a Moh's Surgery program. We will be hiring an experienced Moh's histotech, HT(ASCP) soon. I know the demand for such a position is high. Could anyone give me an idea of the salary range that a histotech gets paid for doing Moh's frozen sections? I would really appreciate it. Wendy Snyder HT(SCP) United Hospital Center Carksburg, WV 26301 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Thu Sep 29 17:24:10 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Sep 29 17:24:36 2005 Subject: [Histonet] what is mohs? Message-ID: I am going to this meeting this year in Atlanta Robyn >>> "Manager" 9/29/2005 12:35 PM >>> The Mohs technique is used primarily for skin cancer, and done using frozen sections. There is a society called the American Society for Mohs Histotechnology. No certification at this time specifically for Mohs, although it is always being talked about. Beth Seattle Skin Cancer Center -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Thursday, September 29, 2005 11:55 AM To: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] what is mohs? It is particular technique that was invented by a man named Frederick Mohs. The technique is that a surgeon is technically the pathologist reading his/her own slides while the patient waits, whereas normally a pathologist would read the slide. And the other unique thing about this particular surgery is that you cut the tissue tangential and not breadloafed. Robyn OHSU >>> "Gudrun Lang" 9/29/2005 10:59 AM >>> Hi, I don't know this term "mohs". Please would someone be so kind to explain. Thank you Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcolclefa <@t> aol.com Wed Sep 28 17:49:55 2005 From: jcolclefa <@t> aol.com (John PJ Coleman) Date: Thu Sep 29 17:37:14 2005 Subject: [Histonet] IHC H2O2, fatty things, IHC slides In-Reply-To: <200509281301.824433acc8428d@rly-yj06.mx.aol.com> References: <200509281301.824433acc8428d@rly-yj06.mx.aol.com> Message-ID: <3bf5d1afb1dfdb0df3e1b340e965c1af@aol.com> H2O2 for IHC : I tried the store bought and it sucked for us. Ironically we did this after our peroxide block wasn't working and we discovered that Ventana had relabelled our expensive H2O2 at their warehouse and sold it to us 6 months after its original expiration date, and under the 2 Ventana labels was a third label showing that this was drugstore H2O2 marked up in price a crazy amount. They blamed their drop shipper, then told me their onsite chemist certified the individual bottles (ok, I don't get that line, drop shipper + onsite chemist=not logical statement) After we cancelled that standing order contract and tossed their instrument in the dumpster behind the hospital, we got concentrated H2O2 from Sigma and we make our own 3 % in water with no tween. We do notice a meniscus/surface tension issue on large or fragmented tissue, and we tried using wash buffer instead of Dh2o, we also added Methanol and found the methanol breaks the surface tension but evaporates too quickly. We went back to just dh2o at 300 ul per slide for 10 min and we don't notice compromised results. We do follow the peroxide with a casein protein block and our slides are clean. We do up to 500 IHC slides a day (yes 500 slides a day max. usually 250 - 300 though) Fatty tissues: We start with a fixative, 10%NBF with ethanol and glacial acetic acid added to fix through the fat a little better than NBF alone. Processor programs have longer 100% ethanols and longer xylene times for better dehydration and clearing. All solutions at 40C. IHC slides: understand the tissue adherence will be influenced down the line by heat retrieval, ph of your retrieval solutions, whether or not you use enzyme digestion, and if you blue your slides in ammonia water. We specifically had problems on the slides with the X on em (I don't have all the brands in my head here at starbucks) when the IHC slides were precut in the main histo lab on water baths with gelatin or Sta-on liquid. We have had no issues with the slides with the + or the P, or the barrier slides from Biogenex. We routinely cut all our slides on a tap water bath with no additives. Email me for details if you have questions on any of the above. On Sep 28, 2005, at 1:02 PM, histonet-request@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Methyl Green (C.M. van der Loos) 2. Dissolving Alizarin complexon (L.Driessen@orthop.umcn.nl) 3. RE: fatty tissue processing (Rogerson Kemlo (ELHT) Pathology) 4. Superfrost Plus slides (Cathy Malcontenti-Wilson) 5. RE: Hydrogen Peroxide for IHC (Lee & Peggy Wenk) 6. RE: haemoxygenase I or II (Edwards, R.E.) 7. SEM on mouse tissue (Stefano Mantero) 8. RE: Superfrost Plus slides (Smith, Jeffery D. (HSC)) 9. RE: Superfrost Plus slides (mucram11@comcast.net) 10. Source of DIFCO trypsin in the UK (Stephen.Eyres@sanofi-aventis.com) 11. Re: processing skin and intestines (Gayle Callis) 12. RE: Frozen tissue washing from slide (DeBrosse_Beatrice) 13. Re: Dissolving Alizarin complexon (Gayle Callis) 14. Pap Pen Blues (Andrea T. Hooper) 15. Re: Superfrost Plus slides (Gayle Callis) 16. PAS staining RE: [Histonet] Frozen tissue washing from slide (Gayle Callis) 17. Re: Pap Pen Blues (Gayle Callis) 18. plastic sections (hstevens@ucsd.edu) 19. Use of methanol and DMSO in Whole mount immunostaining (Johnson, Teri) ---------------------------------------------------------------------- Message: 1 Date: Wed, 28 Sep 2005 08:55:54 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: Methyl Green To: histonet@lists.utsouthwestern.edu Cc: MEeva@mednet.ucla.edu Message-ID: <950f24952f1d.952f1d950f24@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Hi Mervi, The brown-red color combination you mentioned allows a simple hematoxilin counterstain. I would advise you to work with a diluted hematoxilin solution and keep the time short. Check out the counterstaining result microscopically and if needed repeat.You don't want intensely blue nuclei as this will mask your double staining. If you really wish to counterstain with methylgreen soak your slides in 100 mM Na-acetate buffer pH 5.2 for 15 min. Don't wash. Cover sections for 5 min with 0.1% methylgreen (Sigma M6776) dissolved in the acetate buffer as above. Wash briefly with with distilled water and dry the slides completely at a hot plate. Mount organically with VectaMount. This procedure circumvents dehydration by alcohols, which isn't good for your red alk. phos. reaction product. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Tue, 27 Sep 2005 14:19:05 -0700 From: "Eeva, Mervi" Subject: [Histonet] Methyl Green To: histonet@lists.utsouthwestern.edu Hi All, I tried Methyl Green counterstaining on my double stained immunohistochemistry slides and didn't get any visible counterstaining. Tissue was FFPE brain and otherwise I used mostly Vector reagents with DAB and AP-red detection. Any ideas what went wrong? - Mervi Eeva, UCLA ------------------------------ Message: 2 Date: Wed, 28 Sep 2005 09:46:45 +0200 From: Subject: [Histonet] Dissolving Alizarin complexon To: Message-ID: <2E2AC813A84E054C8E8E572131082BE2145510@umcnet14.umcn.nl> Content-Type: text/plain; charset="iso-8859-1" Hello, Is there anyone who is familiar with dissolving alizarin complexon for bonelabeling? We inject this fluorochrome in our testanimals to monitor new boneformation. Because of that we need to sterilize the solution. For this we push the solution through a 0,2 um millipore-filter. The problem is that the filter gets clogged very fast. We've tried prefiltering through a paperfilter, but that doesn't solve the problem. Does anyone have a a solution for this? Greetings, L?on Driessen, Orthopaedic Research Lab UMCN St. Radboud, Nijmegen, The Netherlands ------------------------------ Message: 3 Date: Wed, 28 Sep 2005 09:05:12 +0100 From: "Rogerson Kemlo \(ELHT\) Pathology" Subject: RE: [Histonet] fatty tissue processing To: "Jennifer N. Cresor" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD58B@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" This question is a regular; I don't have a protocol as I am a Manager/ Cytologist but from memory fatty tissues are best cut thin, well fixed, dehydrated and then put into a clearing agent/ degreasant. After the fat has been 'removed' return to dyhydration, then 'clear' again and impregnate with wax. You can use many degreasants but I used xylene, Mr. Lillie 'waxes' eloquently about such matters. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer N. Cresor Sent: Tuesday, September 27, 2005 5:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fatty tissue processing Hello, We are currently in the process of setting up a processor specifically for processing fatty tissues. I would really like to get people's protocols on processing fatty tissues that have worked well for them. Thank you, Jennifer jcresor@icpath.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Wed, 28 Sep 2005 18:33:11 +1000 From: Cathy Malcontenti-Wilson Subject: [Histonet] Superfrost Plus slides To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.1.2.2.20050928182518.02658550@mail.staff.unimelb.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear All, OK so question regarding the superfrost slides. I use them for immunostaining and I remember a few years ago when cutting sections and orientating them in the waterbath, if I got it wrong well bad luck because the sections were well and truly stuck within a few seconds. But now I can orientate them no problem, they lift off easily. Even though they look like most of the sections are still attached, I am getting all this strange background immunostaining around the edges of the tissues and sometimes it looks as though something has been damaging the tissue around the edges as well. I remember a while ago there was some discussion on superfrost slides on histonet but I cant remember what the conclusion was. Has anyone noticed any changes with the superfrost slides, or do they recommend one brand over another? We use Menzel brand and have tried two suppliers, and both the superfrost and the superfrost plus slides. Thanks in advance. Cathy ------------------------------ Message: 5 Date: Wed, 28 Sep 2005 04:53:36 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] Hydrogen Peroxide for IHC To: Message-ID: <1084225373-373221435@pathology.swmed.edu> Content-Type: text/plain; charset="us-ascii" We use "store bought". However, we buy it through our hospital pharmacy, so it comes in fresh and cheaper than off the drug store shelf. Plus, we look at the expiration date (which is always a year down the road). Peggy Wenk.HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, September 27, 2005 5:10 PM To: Poteete, Jacquie A.; 'CRME (Criss Meligro)'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Hydrogen Peroxide for IHC Our lab we always used 3% hydrogen peroxide manufactured by Fisher. We were not willing to pay for "brand" 3% H202 like Ventana. Rene J. "Poteete, Jacquie A." wrote: I don't trust drug store Hydrogen Peroxide. You never know how long it has been on the shelf or how long it had been sitting in a hot warehouse. Jacquie Poteete MT(ASCP)QIHC Lead technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: CRME (Criss Meligro) [mailto:meligroc@zgi.com] Sent: Tuesday, September 27, 2005 2:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hydrogen Peroxide for IHC Just wondering if anyone has purchased 3% Hydrogen Peroxide in the 500 ml bottle from the local drug store at $1.00 +/- and used it for IHC instead of the $66 bottle from Ventana or similar company? Do you add tween? thanks....Criss _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 28 Sep 2005 10:59:03 +0100 From: "Edwards, R.E." Subject: [Histonet] RE: haemoxygenase I or II To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Wanted antibodies to the above that work on mouse paraffin processed tissues...thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K..... ------------------------------ Message: 7 Date: Wed, 28 Sep 2005 14:47:23 +0200 (CEST) From: Stefano Mantero Subject: [Histonet] SEM on mouse tissue To: histonet@lists.utsouthwestern.edu Message-ID: <20050928124723.41442.qmail@web25903.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I need Histo help: I?m looking for a protocol for a Scanning Electron Microscopy sample preparation on embryonal mouse tissue. Please help ! Thanks in advance Stefano --------------------------------- Yahoo! Mail: gratis 1GB per i messaggi, antispam, antivirus, POP3 ------------------------------ Message: 8 Date: Wed, 28 Sep 2005 09:07:15 -0500 From: "Smith, Jeffery D. \(HSC\)" Subject: RE: [Histonet] Superfrost Plus slides To: "Cathy Malcontenti-Wilson" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" We are using VWR Superfrost Plus. We do hundreds of Immunos per week and have had no such problems. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Cathy Malcontenti-Wilson Sent: Wed 9/28/2005 3:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Superfrost Plus slides Dear All, OK so question regarding the superfrost slides. I use them for immunostaining and I remember a few years ago when cutting sections and orientating them in the waterbath, if I got it wrong well bad luck because the sections were well and truly stuck within a few seconds. But now I can orientate them no problem, they lift off easily. Even though they look like most of the sections are still attached, I am getting all this strange background immunostaining around the edges of the tissues and sometimes it looks as though something has been damaging the tissue around the edges as well. I remember a while ago there was some discussion on superfrost slides on histonet but I cant remember what the conclusion was. Has anyone noticed any changes with the superfrost slides, or do they recommend one brand over another? We use Menzel brand and have tried two suppliers, and both the superfrost and the superfrost plus slides. Thanks in advance. Cathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Wed, 28 Sep 2005 14:24:09 +0000 From: mucram11@comcast.net Subject: RE: [Histonet] Superfrost Plus slides To: "Smith, Jeffery D. (HSC)" , "Cathy Malcontenti-Wilson" Cc: histonet@lists.utsouthwestern.edu Message-ID: <092820051424.17323.433AA7890005BBA8000043AB2207001641CECE030E9D0C9A03@c omcast.net> Content-Type: text/plain Cathy, You should also be sure you are roatating your slides so the oldest lot numbers are used first. The coatings are not permenant and can lose surface attraction over time and with exposure to light, heat and air. The expiration date is from the time they are manufactured, not when they arrive to you. Be sure your supplier is sending you fresh slides that have not been sitting in a warehouse for months with varying temperatures and conditions. Hope this helps. Pam Marcum -------------- Original message -------------- > We are using VWR Superfrost Plus. We do hundreds of Immunos per week > and have > had no such problems. > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Cathy > Malcontenti-Wilson > Sent: Wed 9/28/2005 3:33 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Superfrost Plus slides > > > > Dear All, > OK so question regarding the superfrost slides. > I use them for immunostaining and I remember a few years ago when > cutting > sections and orientating them in the waterbath, if I got it wrong well > bad > luck because the sections were well and truly stuck within a few > seconds. > But now I can orientate them no problem, they lift off easily. > Even though they look like most of the sections are still attached, I > am > getting all this strange background immunostaining around the edges of > the > tissues and sometimes it looks as though something has been damaging > the > tissue around the edges as well. I remember a while ago there was some > discussion on superfrost slides on histonet but I cant remember what > the > conclusion was. > Has anyone noticed any changes with the superfrost slides, or do they > recommend one brand over another? > We use Menzel brand and have tried two suppliers, and both the > superfrost > and the superfrost plus slides. > Thanks in advance. > Cathy > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Wed, 28 Sep 2005 15:41:56 +0100 From: Stephen.Eyres@sanofi-aventis.com Subject: [Histonet] Source of DIFCO trypsin in the UK To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi, Could someone give me a source for DIFCO trypsin (0152 or 21530-product code change???)? Many thanks Steve ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- ------------------------------ Message: 11 Date: Wed, 28 Sep 2005 09:00:08 -0600 From: Gayle Callis Subject: Re: [Histonet] processing skin and intestines To: Christian Franci , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050928085212.01b7f430@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Isn't the Citadel a carousel (sp?) style with vacuum on last station of paraffin? I am not totally familiar with this processor. With our VIP (vacuum/pressure throughout), 45 minutes per station of dehyrant, clearing and paraffin (60C) at most but we get superb exchange of solvents with VIP processing, and we never add heat to dehydration/clearing steps - dries out the tissue even more. For rat, I would do 1 hour on VIP. If you do not have vacuum and pressure throughout for each station, then mouse tissue will survive 1 hour timing on a carousel style nicely. Keep in mind animal are less fatty than human, and can easily be overdehydrated, dried out. At 05:38 PM 9/27/2005, you wrote: > Howdy folks! > We've been asked by some scientists to process skin, cheek pouches and > intestines from mice for FF/paraffin embedding for IHC. > I'm just wondering if any of you happen to have a good protocol for > doing > so. I know that for rat tissues one tends to use longer processing > times.. > We just got a Citadel processing machine and I'd like to set up a > program > for this. > thanks a bunch in advance! > > C Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 12 Date: Wed, 28 Sep 2005 08:01:23 -0700 From: "DeBrosse_Beatrice" Subject: RE: [Histonet] Frozen tissue washing from slide To: , Message-ID: Content-Type: text/plain; charset="us-ascii" How long do you air dry the slides? Have you tried to fix the slides in acetone? Is the 10% formalin cold when you fix the slides? How thick are the sections? I really don't see a problem with the Periodic acid, the Schiffs or the water rinses, but I believe your acid solution is too concentrated after your counterstaining. Have you tried 1% acid/alcohol (70% alcohol)? Sincerely, Beatrice DeBrosse-Serra BS, HT(ASCP) Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of WWmn916@aol.com Sent: Tuesday, September 27, 2005 6:01 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Frozen tissue washing from slide I'm hoping someone can give me some idea why frozen muscle sections would suddenly start washing away from slide while trying to do PAS w/wo stain? Other muscle panel stains do okay in staining process.....just the PAS w/wo is recently having problems. -We use superfrost plus slides -Sections are fixed in 10% formalin prior to stain -Most water rinses are DI. Only one tap water rinse step right after amylase digest. -Does periodic acid have to be room temp prior to staining? -Could Shiffs reagent be too cold? -does temperature of water rinses affect adherance of sections to slides? -We use a 10% acid water rinse after heme staining. I recall if acid water is made incorrectly, tissue sections could wash much like they do if acid water in H+E stain is too strong. Thanks for any thoughts and help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Wed, 28 Sep 2005 09:08:59 -0600 From: Gayle Callis Subject: Re: [Histonet] Dissolving Alizarin complexon To: , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050928090319.01b71a78@gemini.msu.montana.edu> Content-Type: text/plain; charset="iso-8859-1"; format=flowed Have you tried a larger pore size, 0.45 um.or larger. One could always get the big particles first, then refilter again through smaller pore size if you felt it necessary. I have had to do this with normal serums before when 0.22um clogs up. Possibly, you could try centrifuging the stock solution, decant off the top what you need and microfilter that portion just to eliminate the biggest particles before you go to microfilter. Just suggestions here At 01:46 AM 9/28/2005, you wrote: > Hello, > > Is there anyone who is familiar with dissolving alizarin complexon for > bonelabeling? > We inject this fluorochrome in our testanimals to monitor new > boneformation. Because of that we need to sterilize the solution. For > this > we push the solution through a 0,2 um millipore-filter. The problem is > that the filter gets clogged very fast. We've tried prefiltering > through a > paperfilter, but that doesn't solve the problem. Does anyone have a a > solution for this? > > Greetings, > > L?on Driessen, > Orthopaedic Research Lab > UMCN St. Radboud, Nijmegen, > The Netherlands > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 14 Date: Wed, 28 Sep 2005 11:29:57 -0400 From: "Andrea T. Hooper" Subject: [Histonet] Pap Pen Blues To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Hello All, I have used Zymed Mini Pap pens for years now as through trials and tribulations and talking with coworkers we agreed it was the best out there. The pen rarely came up or leaked and gave beautiful results. Even withstood heat retrieval in DAKO Target Retrieval Solution with no problems. The stuff was indestructible!! This is until recently ... I have noticed with the past several orders that the formula has changed. The pap pen "ink" is now a blue green in color (used to be more generic yellow/beige) and it does not stay on well at all. I end up going home in a state of frustration every day as I pap pen hundreds of slides with little success for the entirety of my experiment. Arggggh! So I have two questions: (1) Has anyone else noticed this change in formula over the past year or so? (It may have been longer as I was working with a stock of pap pens before that) (2) Who makes your favorite pap pen? THANKS! Andrea -- ------------------------------ Message: 15 Date: Wed, 28 Sep 2005 09:29:10 -0600 From: Gayle Callis Subject: Re: [Histonet] Superfrost Plus slides To: Cathy Malcontenti-Wilson, Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050928090945.01b54cf0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Cathy, A couple of things, for reorientation of sections, we use the Erie Polysine slides (poly l lysine coating) that lets us manipulate sections on slide at waterbath. Plus Charge are silane and once the section is attracted to slide surface, you are done! Do you use Tween or a detergent in your buffers/diluents before chromogen application and throughout the staining procedure? If not what you may be experiencing is hydrophobic bonding interactions between tissue and reagent proteins (BSA, serums) and ionic interaction of chromogen to slide surface, which the detergent will eliminate giving cleaner results. 0.05% Tween 20 is usually adequate to eliminate this problem. We have used Erie's SuperFrost Plus Charge, their trademark and also Polysine for section manipulation problems for years, and love them. We have used Plus charge slides stored long past expiration date without any problems with IHC or other routine staining. What other manufacturers do for coating slides may vary from Erie products, one can only try them out to see if they work well. As for tissue section damage, can you describe in further detail please? I doubt this is caused by the slide coatings but some other problem. At 02:33 AM 9/28/2005, you wrote: > Dear All, > OK so question regarding the superfrost slides. > I use them for immunostaining and I remember a few years ago when > cutting > sections and orientating them in the waterbath, if I got it wrong well > bad > luck because the sections were well and truly stuck within a few > seconds. > But now I can orientate them no problem, they lift off easily. > Even though they look like most of the sections are still attached, I > am > getting all this strange background immunostaining around the edges of > the > tissues and sometimes it looks as though something has been damaging > the > tissue around the edges as well. I remember a while ago there was some > discussion on superfrost slides on histonet but I cant remember what > the > conclusion was. > Has anyone noticed any changes with the superfrost slides, or do they > recommend one brand over another? > We use Menzel brand and have tried two suppliers, and both the > superfrost > and the superfrost plus slides. > Thanks in advance. > Cathy > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 16 Date: Wed, 28 Sep 2005 09:46:26 -0600 From: Gayle Callis Subject: PAS staining RE: [Histonet] Frozen tissue washing from slide To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050928093504.01b8d3b8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed With frozen sections, a gentle rinse is advisable. We cut frozen sections destined for routine stains and go directly to room temperature NBF - air drying is not really necessary for routine stains but is for immunostaiing. Let your frozen sections fix longer than 10 minutes to ensure good adherance to slide and fixed proteins. You can proceed with a PAS stain just as you do a paraffin section, at room temperature. Is the 10% acid glacial acetic or hydrochloric acid? You did not specify which one. If you do acid/alcohol rinse, 0.5 to 1% HCl in 70% for regressive or Harris hematoxylin , or if you use acetic acid, 4% in water is sufficient with progressive Gill type hematoxylin. > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > WWmn916@aol.com > Sent: Tuesday, September 27, 2005 6:01 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] Frozen tissue washing from slide > > I'm hoping someone can give me some idea why frozen muscle sections > would > suddenly start washing away from slide while trying to do PAS w/wo > stain? > Other muscle panel stains do okay in staining process.....just the PAS > w/wo is > recently having problems. > -We use superfrost plus slides > -Sections are fixed in 10% formalin prior to stain > -Most water rinses are DI. Only one tap water rinse step right after > amylase digest. > -Does periodic acid have to be room temp prior to staining? > -Could Shiffs reagent be too cold? > -does temperature of water rinses affect adherance of sections to > slides? > -We use a 10% acid water rinse after heme staining. I recall if acid > water > is made incorrectly, tissue sections could wash much like they do if > acid > water in H+E stain is too strong. > > Thanks for any thoughts and help. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 17 Date: Wed, 28 Sep 2005 10:04:23 -0600 From: Gayle Callis Subject: Re: [Histonet] Pap Pen Blues To: "Andrea T. Hooper" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050928095641.01b98148@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Andrea, We now use Vectors ImmEdge pens - doesn't come up, and has a hint of color. Cheaper, two to a set. Can remark a slide when wet, although we do blot away moisture, but the pen goo does not lift nor flow across a section if it contacts buffer - been there and have experienced the "Pap pen oil slick syndrome" that ruins IHC when section gets goo-coated. To use ImmEdge, be sure to shake pens very vigorously to redistribute the goo (per their instructions). We vortex them hard to do that job. Maybe the pens you have now need a good vortexing - worth a try for pricey little pens. Pressing the point to start goo is still something that requires gentle tough, but I have not had too many problems with that. Flow is generally excellent, right amount. At 09:29 AM 9/28/2005, you wrote: > Hello All, > > I have used Zymed Mini Pap pens for years now as through trials and > tribulations and talking with coworkers we agreed it was the best out > there. The pen rarely came up or leaked and gave beautiful results. > Even > withstood heat retrieval in DAKO Target Retrieval Solution with no > problems. The stuff was indestructible!! > > This is until recently ... I have noticed with the past several orders > that the formula has changed. The pap pen "ink" is now a blue green in > color (used to be more generic yellow/beige) and it does not stay on > well > at all. I end up going home in a state of frustration every day as I > pap > pen hundreds of slides with little success for the entirety of my > experiment. Arggggh! > > So I have two questions: > > (1) Has anyone else noticed this change in formula over the past year > or > so? (It may have been longer as I was working with a stock of pap pens > before that) > (2) Who makes your favorite pap pen? > > THANKS! > Andrea > -- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 18 Date: Wed, 28 Sep 2005 09:20:45 -0700 (PDT) From: hstevens@ucsd.edu Subject: [Histonet] plastic sections To: histonet@lists.utsouthwestern.edu Message-ID: <32021.67.113.110.37.1127924445.squirrel@acs-webmail.ucsd.edu> Content-Type: text/plain;charset=iso-8859-1 Hi, I am looking for a good inexpensive lab that can process mouse femurs (at present in alcohol) to obtain plastic embedded midshaft sections (30 um ish). Does anyone have any recommendations for the San Diego area or beyond? Thanks Hazel Stevens ------------------------------ Message: 19 Date: Wed, 28 Sep 2005 11:53:22 -0500 From: "Johnson, Teri" Subject: [Histonet] Use of methanol and DMSO in Whole mount immunostaining To: Message-ID: Content-Type: text/plain; charset="us-ascii" Ref: GENES & DEVELOPMENT 9:2509-2522, Misexpression of Hoxa-13 induces cartilage homeotic transformation and changes cell adhesiveness in chick limb buds, Yokouchi, et al In this study, chick embryos were dissected and fixed in methanol/DMSO (4:1) at 4 degrees C overnight, followed by peroxidase blocking in 5% hydrogen peroxide in methanol/DMSO (4:1). I am unfamiliar with using methanol/DMSO as a tissue fixative for whole mount studies. This can obviously be quite helpful for finding protein targets that are masked or made unrecognizable using aldehyde fixatives. What effect would the DMSO have on the samples prior to immunostaining? Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 22, Issue 37 **************************************** JPJC-757 335-2159 http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ From cbass <@t> bidmc.harvard.edu Thu Sep 29 17:59:21 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Thu Sep 29 17:59:47 2005 Subject: [Histonet] fluorescent proteins (RFP and GFP) native fluorescence Message-ID: <61F5077F-B2B2-450C-AC41-3B4B0399703C@bidmc.harvard.edu> Hey guys, I was hoping someone here could give me some advice on their experience with various fluorescent proteins. I need a good marker for my viral vector. I have a humanized GFP, RFP and EGFP. I would like to inject the vector into the tail vein to see which tissue lights up. My hGFP variant is not very strong in vitro, but both RFP and EGFP work well. I have heard that GFP quickly disperses when mounted on slides. However, I have seen some papers that use native EGFP fluorescence with cryosections and they don't have problems. Ideally I would use floating tissue sections of 40 microns as I have easy access to a sliding microtome. I have heard that RFP does not work with fixation at all. I am open to any sort of advice. Could someone recommend a protocol for visualizing native fluorescence with either EGFP or RFP? Specifically, whether to fix or not, the thickness of the section, floating sections vs. slide mounts, etc. Also, if there is another fluorescent protein available that you could suggest I would like to hear about it. I have heard of yellow and blue fluorescent proteins as well as a monomeric form or RFP. I am also open to using a "universal" fusion protein, for example actin- GFP if it solves the problem of GFP native fluorescence. Thanks, Caroline From xli <@t> amgen.com Thu Sep 29 18:09:46 2005 From: xli <@t> amgen.com (Li, Xiaodong) Date: Thu Sep 29 18:10:10 2005 Subject: [Histonet] Associate position at Amgen Message-ID: Hello, I would like to send following message to the members: We are seeking an experienced and motivated Research Associate in our drug discovery programs. The successful candidate will utilize bone histomorphometry methods to support drug discovery research and should have strong laboratory skills and expertise in histology techniques. The qualified candidate will have 5 or more years of experience and a Bachelors or Masters degree in a relevant scientific discipline. The individual should be able to work independently, and as part of a research project team. Candidates with experience in data analysis and data management, who have excellent problem solving abilities and very good communication skills are encouraged to apply. Under the limited supervision, he/she will perform histomorphometry analysis. Other job responsibilities include maintenance of careful records of all laboratory and animal experiments, histomorphometry sample preparation. Industrial experience is preferred. Please send the resume to xli@amgen.com From arvind <@t> nbrc.res.in Thu Sep 29 23:36:02 2005 From: arvind <@t> nbrc.res.in (Arvind) Date: Thu Sep 29 23:32:47 2005 Subject: [Histonet] querry References: Message-ID: <004501c5c578$7a6f5070$aa00a8c0@nbrc.res.in> has anyone done DopamineD2 recptor on human fetal brain or adult human brain if yes please can you share your protocol thanking in advance arvind singh pundir national brain research centre gurgaon,India From christelle.gerard <@t> novartis.com Fri Sep 30 04:29:32 2005 From: christelle.gerard <@t> novartis.com (christelle.gerard@novartis.com) Date: Fri Sep 30 04:29:56 2005 Subject: [Histonet] antibody aginst cathepsin S Message-ID: Hi All, I wanted to run an IHC against Cathepsin S on frozen sections rat tissue . Does anyone have experience with antibody from Santa Cruz (sc 6505) or an other for this IHC. If so, please recommend some protocols, some antibody. Thanks in advance. Christelle From angela.mcnabola.b <@t> bayer.com Fri Sep 30 07:25:24 2005 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Fri Sep 30 07:25:26 2005 Subject: [Histonet] Tissue fixation help Message-ID: Good Morning, Does anybody on the histonet have any experience with taking tissue that has been snap frozen in minus 80, and then "thawed" out and formalin fixed and paraffin embedded? I won't get into the reasons why we have to do this, but does anyone have a protocol (or just slowly thaw it out and fix it??). How did your IHC turn out, especially when using phosphorlyated antibodies? Did you get good morphology of your tissue? thanks in advance! Angela Angela McNabola, MS HT(ASCP)SLS, QIHC Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 203-812-5001 angela.mcnabola.b@bayer.com From MSafron <@t> wilresearch.com Fri Sep 30 08:09:42 2005 From: MSafron <@t> wilresearch.com (Mike Safron) Date: Fri Sep 30 08:13:34 2005 Subject: [Histonet] for anyone in pharmaceutical, contract labs, etc Message-ID: We typically have one organ weigher. Michael Safron A.S.,HT(ASCP) Manager, Histology WIL Research Laboratories LLC 419-289-8700 ext: 3040 Ashland, Ohio 44805 From rjbuesa <@t> yahoo.com Fri Sep 30 08:21:47 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Sep 30 08:22:09 2005 Subject: [Histonet] IHC H2O2, fatty things, IHC slides In-Reply-To: <3bf5d1afb1dfdb0df3e1b340e965c1af@aol.com> Message-ID: <20050930132147.57869.qmail@web61217.mail.yahoo.com> Why I am not surprised about what Ventana did? What took you so long to dump the Ventana instrument? We always used commercial 3% (Fisher) H202 but never used pure H202 to dilute (too dangerous, too many restrictions, too expensive, too problematic!). We never added Tween to H202; what we did in order to cut time with our Dako autostainer, was to do the peroxidase block step outside the instrument. After HIER the slides to PBS/Tween; to a container with 3% H202 for 5 minutes (being totally immesed in 3% H202, Tween was not necessary!); to PBS/Tween to the instrument to start the run with the primary antibody. This way we cut @ 25 min. from each 48 slides run and had no problems at all. Rene J. John PJ Coleman wrote: H2O2 for IHC : I tried the store bought and it sucked for us. Ironically we did this after our peroxide block wasn't working and we discovered that Ventana had relabelled our expensive H2O2 at their warehouse and sold it to us 6 months after its original expiration date, and under the 2 Ventana labels was a third label showing that this was drugstore H2O2 marked up in price a crazy amount. They blamed their drop shipper, then told me their onsite chemist certified the individual bottles (ok, I don't get that line, drop shipper + onsite chemist=not logical statement) After we cancelled that standing order contract and tossed their instrument in the dumpster behind the hospital, we got concentrated H2O2 from Sigma and we make our own 3 % in water with no tween. We do notice a meniscus/surface tension issue on large or fragmented tissue, and we tried using wash buffer instead of Dh2o, we also added Methanol and found the methanol breaks the surface tension but evaporates too quickly. We went back to just dh2o at 300 ul per slide for 10 min and we don't notice compromised results. We do follow the peroxide with a casein protein block and our slides are clean. We do up to 500 IHC slides a day (yes 500 slides a day max. usually 250 - 300 though) Fatty tissues: We start with a fixative, 10%NBF with ethanol and glacial acetic acid added to fix through the fat a little better than NBF alone. Processor programs have longer 100% ethanols and longer xylene times for better dehydration and clearing. All solutions at 40C. IHC slides: understand the tissue adherence will be influenced down the line by heat retrieval, ph of your retrieval solutions, whether or not you use enzyme digestion, and if you blue your slides in ammonia water. We specifically had problems on the slides with the X on em (I don't have all the brands in my head here at starbucks) when the IHC slides were precut in the main histo lab on water baths with gelatin or Sta-on liquid. We have had no issues with the slides with the + or the P, or the barrier slides from Biogenex. We routinely cut all our slides on a tap water bath with no additives. Email me for details if you have questions on any of the above. On Sep 28, 2005, at 1:02 PM, histonet-request@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Methyl Green (C.M. van der Loos) 2. Dissolving Alizarin complexon (L.Driessen@orthop.umcn.nl) 3. RE: fatty tissue processing (Rogerson Kemlo (ELHT) Pathology) 4. Superfrost Plus slides (Cathy Malcontenti-Wilson) 5. RE: Hydrogen Peroxide for IHC (Lee & Peggy Wenk) 6. RE: haemoxygenase I or II (Edwards, R.E.) 7. SEM on mouse tissue (Stefano Mantero) 8. RE: Superfrost Plus slides (Smith, Jeffery D. (HSC)) 9. RE: Superfrost Plus slides (mucram11@comcast.net) 10. Source of DIFCO trypsin in the UK (Stephen.Eyres@sanofi-aventis.com) 11. Re: processing skin and intestines (Gayle Callis) 12. RE: Frozen tissue washing from slide (DeBrosse_Beatrice) 13. Re: Dissolving Alizarin complexon (Gayle Callis) 14. Pap Pen Blues (Andrea T. Hooper) 15. Re: Superfrost Plus slides (Gayle Callis) 16. PAS staining RE: [Histonet] Frozen tissue washing from slide (Gayle Callis) 17. Re: Pap Pen Blues (Gayle Callis) 18. plastic sections (hstevens@ucsd.edu) 19. Use of methanol and DMSO in Whole mount immunostaining (Johnson, Teri) ---------------------------------------------------------------------- Message: 1 Date: Wed, 28 Sep 2005 08:55:54 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: Methyl Green To: histonet@lists.utsouthwestern.edu Cc: MEeva@mednet.ucla.edu Message-ID: <950f24952f1d.952f1d950f24@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Hi Mervi, The brown-red color combination you mentioned allows a simple hematoxilin counterstain. I would advise you to work with a diluted hematoxilin solution and keep the time short. Check out the counterstaining result microscopically and if needed repeat.You don't want intensely blue nuclei as this will mask your double staining. If you really wish to counterstain with methylgreen soak your slides in 100 mM Na-acetate buffer pH 5.2 for 15 min. Don't wash. Cover sections for 5 min with 0.1% methylgreen (Sigma M6776) dissolved in the acetate buffer as above. Wash briefly with with distilled water and dry the slides completely at a hot plate. Mount organically with VectaMount. This procedure circumvents dehydration by alcohols, which isn't good for your red alk. phos. reaction product. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 Date: Tue, 27 Sep 2005 14:19:05 -0700 From: "Eeva, Mervi" Subject: [Histonet] Methyl Green To: histonet@lists.utsouthwestern.edu Hi All, I tried Methyl Green counterstaining on my double stained immunohistochemistry slides and didn't get any visible counterstaining. Tissue was FFPE brain and otherwise I used mostly Vector reagents with DAB and AP-red detection. Any ideas what went wrong? - Mervi Eeva, UCLA ------------------------------ Message: 2 Date: Wed, 28 Sep 2005 09:46:45 +0200 From: Subject: [Histonet] Dissolving Alizarin complexon To: Message-ID: <2E2AC813A84E054C8E8E572131082BE2145510@umcnet14.umcn.nl> Content-Type: text/plain; charset="iso-8859-1" Hello, Is there anyone who is familiar with dissolving alizarin complexon for bonelabeling? We inject this fluorochrome in our testanimals to monitor new boneformation. Because of that we need to sterilize the solution. For this we push the solution through a 0,2 um millipore-filter. The problem is that the filter gets clogged very fast. We've tried prefiltering through a paperfilter, but that doesn't solve the problem. Does anyone have a a solution for this? Greetings, L?on Driessen, Orthopaedic Research Lab UMCN St. Radboud, Nijmegen, The Netherlands ------------------------------ Message: 3 Date: Wed, 28 Sep 2005 09:05:12 +0100 From: "Rogerson Kemlo \(ELHT\) Pathology" Subject: RE: [Histonet] fatty tissue processing To: "Jennifer N. Cresor" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD58B@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" This question is a regular; I don't have a protocol as I am a Manager/ Cytologist but from memory fatty tissues are best cut thin, well fixed, dehydrated and then put into a clearing agent/ degreasant. After the fat has been 'removed' return to dyhydration, then 'clear' again and impregnate with wax. You can use many degreasants but I used xylene, Mr. Lillie 'waxes' eloquently about such matters. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer N. Cresor Sent: Tuesday, September 27, 2005 5:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fatty tissue processing Hello, We are currently in the process of setting up a processor specifically for processing fatty tissues. I would really like to get people's protocols on processing fatty tissues that have worked well for them. Thank you, Jennifer jcresor@icpath.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Wed, 28 Sep 2005 18:33:11 +1000 From: Cathy Malcontenti-Wilson Subject: [Histonet] Superfrost Plus slides To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.1.2.2.20050928182518.02658550@mail.staff.unimelb.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear All, OK so question regarding the superfrost slides. I use them for immunostaining and I remember a few years ago when cutting sections and orientating them in the waterbath, if I got it wrong well bad luck because the sections were well and truly stuck within a few seconds. But now I can orientate them no problem, they lift off easily. Even though they look like most of the sections are still attached, I am getting all this strange background immunostaining around the edges of the tissues and sometimes it looks as though something has been damaging the tissue around the edges as well. I remember a while ago there was some discussion on superfrost slides on histonet but I cant remember what the conclusion was. Has anyone noticed any changes with the superfrost slides, or do they recommend one brand over another? We use Menzel brand and have tried two suppliers, and both the superfrost and the superfrost plus slides. Thanks in advance. Cathy ------------------------------ Message: 5 Date: Wed, 28 Sep 2005 04:53:36 -0400 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] Hydrogen Peroxide for IHC To: Message-ID: <1084225373-373221435@pathology.swmed.edu> Content-Type: text/plain; charset="us-ascii" We use "store bought". However, we buy it through our hospital pharmacy, so it comes in fresh and cheaper than off the drug store shelf. Plus, we look at the expiration date (which is always a year down the road). Peggy Wenk.HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, September 27, 2005 5:10 PM To: Poteete, Jacquie A.; 'CRME (Criss Meligro)'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Hydrogen Peroxide for IHC Our lab we always used 3% hydrogen peroxide manufactured by Fisher. We were not willing to pay for "brand" 3% H202 like Ventana. Rene J. "Poteete, Jacquie A." wrote: I don't trust drug store Hydrogen Peroxide. You never know how long it has been on the shelf or how long it had been sitting in a hot warehouse. Jacquie Poteete MT(ASCP)QIHC Lead technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: CRME (Criss Meligro) [mailto:meligroc@zgi.com] Sent: Tuesday, September 27, 2005 2:48 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Hydrogen Peroxide for IHC Just wondering if anyone has purchased 3% Hydrogen Peroxide in the 500 ml bottle from the local drug store at $1.00 +/- and used it for IHC instead of the $66 bottle from Ventana or similar company? Do you add tween? thanks....Criss _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 28 Sep 2005 10:59:03 +0100 From: "Edwards, R.E." Subject: [Histonet] RE: haemoxygenase I or II To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Wanted antibodies to the above that work on mouse paraffin processed tissues...thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K..... ------------------------------ Message: 7 Date: Wed, 28 Sep 2005 14:47:23 +0200 (CEST) From: Stefano Mantero Subject: [Histonet] SEM on mouse tissue To: histonet@lists.utsouthwestern.edu Message-ID: <20050928124723.41442.qmail@web25903.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I need Histo help: I?m looking for a protocol for a Scanning Electron Microscopy sample preparation on embryonal mouse tissue. Please help ! Thanks in advance Stefano --------------------------------- Yahoo! Mail: gratis 1GB per i messaggi, antispam, antivirus, POP3 ------------------------------ Message: 8 Date: Wed, 28 Sep 2005 09:07:15 -0500 From: "Smith, Jeffery D. \(HSC\)" Subject: RE: [Histonet] Superfrost Plus slides To: "Cathy Malcontenti-Wilson" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" We are using VWR Superfrost Plus. We do hundreds of Immunos per week and have had no such problems. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Cathy Malcontenti-Wilson Sent: Wed 9/28/2005 3:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Superfrost Plus slides Dear All, OK so question regarding the superfrost slides. I use them for immunostaining and I remember a few years ago when cutting sections and orientating them in the waterbath, if I got it wrong well bad luck because the sections were well and truly stuck within a few seconds. But now I can orientate them no problem, they lift off easily. Even though they look like most of the sections are still attached, I am getting all this strange background immunostaining around the edges of the tissues and sometimes it looks as though something has been damaging the tissue around the edges as well. I remember a while ago there was some discussion on superfrost slides on histonet but I cant remember what the conclusion was. Has anyone noticed any changes with the superfrost slides, or do they recommend one brand over another? We use Menzel brand and have tried two suppliers, and both the superfrost and the superfrost plus slides. Thanks in advance. Cathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Wed, 28 Sep 2005 14:24:09 +0000 From: mucram11@comcast.net Subject: RE: [Histonet] Superfrost Plus slides To: "Smith, Jeffery D. (HSC)" , "Cathy Malcontenti-Wilson" Cc: histonet@lists.utsouthwestern.edu Message-ID: <092820051424.17323.433AA7890005BBA8000043AB2207001641CECE030E9D0C9A03@c omcast.net> Content-Type: text/plain Cathy, You should also be sure you are roatating your slides so the oldest lot numbers are used first. The coatings are not permenant and can lose surface attraction over time and with exposure to light, heat and air. The expiration date is from the time they are manufactured, not when they arrive to you. Be sure your supplier is sending you fresh slides that have not been sitting in a warehouse for months with varying temperatures and conditions. Hope this helps. Pam Marcum -------------- Original message -------------- > We are using VWR Superfrost Plus. We do hundreds of Immunos per week > and have > had no such problems. > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Cathy > Malcontenti-Wilson > Sent: Wed 9/28/2005 3:33 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Superfrost Plus slides > > > > Dear All, > OK so question regarding the superfrost slides. > I use them for immunostaining and I remember a few years ago when > cutting > sections and orientating them in the waterbath, if I got it wrong well > bad > luck because the sections were well and truly stuck within a few > seconds. > But now I can orientate them no problem, they lift off easily. > Even though they look like most of the sections are still attached, I > am > getting all this strange background immunostaining around the edges of > the > tissues and sometimes it looks as though something has been damaging > the > tissue around the edges as well. I remember a while ago there was some > discussion on superfrost slides on histonet but I cant remember what > the > conclusion was. > Has anyone noticed any changes with the superfrost slides, or do they > recommend one brand over another? > We use Menzel brand and have tried two suppliers, and both the > superfrost > and the superfrost plus slides. > Thanks in advance. > Cathy > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Wed, 28 Sep 2005 15:41:56 +0100 From: Stephen.Eyres@sanofi-aventis.com Subject: [Histonet] Source of DIFCO trypsin in the UK To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi, Could someone give me a source for DIFCO trypsin (0152 or 21530-product code change???)? Many thanks Steve ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- ------------------------------ Message: 11 Date: Wed, 28 Sep 2005 09:00:08 -0600 From: Gayle Callis Subject: Re: [Histonet] processing skin and intestines To: Christian Franci , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050928085212.01b7f430@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Isn't the Citadel a carousel (sp?) style with vacuum on last station of paraffin? I am not totally familiar with this processor. With our VIP (vacuum/pressure throughout), 45 minutes per station of dehyrant, clearing and paraffin (60C) at most but we get superb === message truncated === --------------------------------- Yahoo! for Good Click here to donate to the Hurricane Katrina relief effort. From anh2006 <@t> med.cornell.edu Fri Sep 30 08:49:34 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 30 08:45:17 2005 Subject: [Histonet] CXCR4 and VEGF Receptors Message-ID: Hello All, I am just trying to get a broad feel for what antibodies people are using lately for CXCR4 and VEGF receptors in both human and mouse tissue. Thanks, Andrea -- From ttroyer <@t> petersonlab.com Fri Sep 30 09:35:48 2005 From: ttroyer <@t> petersonlab.com (Travis Troyer) Date: Fri Sep 30 09:36:04 2005 Subject: [Histonet] balanced oxalate Message-ID: <000801c5c5cc$3fe35330$6601010a@Peterson.local> Has anyone ever heard of balanced oxalate for transferring bone marrow aspirates. If so, where can it be purchased. Thanks, Travis Troyer Peterson Laboratory Services Manhattan, KS From mhorne <@t> upei.ca Fri Sep 30 10:40:52 2005 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 30 09:44:31 2005 Subject: [Histonet] Hotel key cards, potential identity theft during travel - sent to our university faculty In-Reply-To: <6.0.0.22.1.20050929151625.01b3d0a8@gemini.msu.montana.edu> Message-ID: <433D2443.23830.5B6EF7@localhost> Anyone interested in this topic might like to go to this site : www.snopes.com/crime/warnings/hotelkey.asp Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From denise <@t> pclv.net Fri Sep 30 10:02:32 2005 From: denise <@t> pclv.net (Denise) Date: Fri Sep 30 10:01:18 2005 Subject: [Histonet] Histology Gross Tech Message-ID: Phoenix Central Laboratory for Veterinarians, Inc. has a position available for an HT or eligible person to perform gross dissection of veterinary tissue specimens. This is a part-time weekend position. Please submit your resume to Denise DeRouchey, c/o Phoenix Central Laboratory, 11620 Airport Road, Everett, WA 98204. Alternately, you may fax your resume to 425-355-3676. Phoenix Central Laboratory, (PCLV), is a veterinary reference laboratory that provides diagnostic services to more than 800 clinics located throughout the Pacific Northwest and Alaska. We are dedicated to providing rapid, quality service. Our laboratory is located in Everett, Washington, just thirty minutes north of downtown Seattle. From cgfields <@t> lexhealth.org Fri Sep 30 10:05:35 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 30 10:03:44 2005 Subject: [Histonet] Silicone slides Message-ID: Hi, I thought I had read something about silicone (coated) slides on the Histo-Net but I may be mistaken. One of our pathologists asked if I I knew where to get silicone coated slides this morning? Have we discussed this and I missed it? No idea yet why he wants this... Thank you in advance for any help..... Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From PMonfils <@t> Lifespan.org Fri Sep 30 10:22:45 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 30 10:23:11 2005 Subject: [Histonet] Silicone slides Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175E8@lsexch.lsmaster.lifespan.org> He is probably referring to what we now call "Plus slides", which are coated with "Silane", a trade name for positively charged silicon tetrahydride. When these slides first came on the market some years ago, the name "Silane" was not yet in vogue, and the maufacturer described the coating on the slides simply as "electrostatically charged silicon". > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Carole Fields > Sent: Friday, September 30, 2005 8:05 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silicone slides > > Hi, > > I thought I had read something about silicone (coated) slides on the > Histo-Net but I may be mistaken. One of our pathologists asked if I I > knew > where to get silicone coated slides this morning? Have we discussed this > and I missed it? No idea yet why he wants this... > Thank you in advance for any help..... > > Carole Fields, HT,ASCP > Pathology Supervisor > Lexington Medical Center > 2720 Sunset Blvd. > W. Columbia, SC 29169 > > > > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From CMCCOLLOUGH <@t> dnr.state.md.us Fri Sep 30 10:34:27 2005 From: CMCCOLLOUGH <@t> dnr.state.md.us (McCollough, Carol) Date: Fri Sep 30 10:34:48 2005 Subject: [Histonet] VIR question - finfish tags Message-ID: Greetings Histonetters: We have had a request from a researcher at Woods Hole for embedding and sectioning of monkfish that have been tagged. The investigator is interested in the interface between the living tissue and the tag. I believe that these are Floy-type spaghetti tags, which are flexible soft plastic, maybe latex. I have no experience with sectioning non-biological materials. We do 99.999% paraffin here, but do have the capability to do methacrylates and resins. Suggestions? Regards - Carol ********************** Carol B. McCollough, HT/HTL(ASCP) Aquatic Animal Research Pathologist Oyster Disease Research Project Fisheries Service Maryland Department of Natural Resources Cooperative Oxford Laboratory 904 S. Morris Street Oxford, Maryland 21654 410-226-5193 x124 From dford <@t> deltapathology.com Fri Sep 30 10:43:19 2005 From: dford <@t> deltapathology.com (D. Ford) Date: Fri Sep 30 10:37:23 2005 Subject: [Histonet] Positions in Louisiana Message-ID: Delta Pathology, in Shreveport Louisiana, currently looking for HT/HTLs. If you have been displaced by the hurricanes and want to return to your home state or if you are looking to relocate please contact us for more information. Darlene Ford, B.S. CT, HT (ASCP) Technical Supervisor 2915 Missouri Avenue Shreveport, LA 71109 318-621-8820 phone 318-671-5922 fax dford@deltapathology.com From ryaskovich <@t> dir.nidcr.nih.gov Fri Sep 30 10:43:26 2005 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 30 10:43:52 2005 Subject: [Histonet] VIR question - finfish tags Message-ID: Carol, Tell them to try Glycol Methacrylate. It works well on fish and plastics. Ruth Yaskovich National Institutes of Health National Institute of Dental and Crainiofacial Research Pain and Neurosensory Mechanism Branch > ---------- > From: McCollough, Carol > Sent: Friday, September 30, 2005 10:34 AM > To: Histonet (E-mail 2) > Subject: [Histonet] VIR question - finfish tags > > Greetings Histonetters: > > We have had a request from a researcher at Woods Hole for embedding and > sectioning of monkfish that have been tagged. The investigator is > interested in the interface between the living tissue and the tag. I > believe that these are Floy-type spaghetti tags, which are flexible soft > plastic, maybe latex. I have no experience with sectioning non-biological > materials. We do 99.999% paraffin here, but do have the capability to do > methacrylates and resins. Suggestions? > > Regards - > Carol > ********************** > Carol B. McCollough, HT/HTL(ASCP) > Aquatic Animal Research Pathologist > Oyster Disease Research Project > Fisheries Service > Maryland Department of Natural Resources > Cooperative Oxford Laboratory > 904 S. Morris Street > Oxford, Maryland 21654 > 410-226-5193 x124 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gtesdall <@t> yahoo.com Fri Sep 30 11:13:37 2005 From: gtesdall <@t> yahoo.com (greg tesdall) Date: Fri Sep 30 11:13:58 2005 Subject: [Histonet] Fisher 172 slide stainer Message-ID: <20050930161337.74096.qmail@web33507.mail.mud.yahoo.com> Hi. Is there a source for parts or a repair consult for the Fisher 172 Histomatic slide stainer. Thanks, Greg Tesdall Kearney, NE. __________________________________ Yahoo! Mail - PC Magazine Editors' Choice 2005 http://mail.yahoo.com From donald <@t> labcareer.com Fri Sep 30 11:30:17 2005 From: donald <@t> labcareer.com (Donald Knowles) Date: Fri Sep 30 11:29:34 2005 Subject: [Histonet] Illinois position Message-ID: <26A7728EF768DA478F45954252293C2037F9EE@matrix.HCCI.local> Senior Histotechnologist position open. The hours would be 9:30/10:30am to 6/7pm Mon-Friday with rotating Saturdays ( definitely a day shift vs early mornings as is typical in histology labs). The Saturday rotation would be about once every 4-5 weeks when we're fully staffed. The applicant must be HT or HTL certified and must meet the CLIA regs to do high complexity testing since they handle/triage/gross renal and neuropath biopsies. I have 25 other positions throughout the country as well. Hope everyone has a great weekend! Don Knowles Healthcare Connections Inc. 866-346-8522 ext 311 From cgfields <@t> lexhealth.org Fri Sep 30 11:36:18 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 30 11:34:21 2005 Subject: [Histonet] Silicone slides Message-ID: I asked him if he meant Silane slides (which we have) and plus slides and he said no he wants Silicone. Maybe he read something that said silicon tetrahydride. I will asked...that is probably it. Thank you for the help.... Carole -----Original Message----- From: Monfils, Paul [mailto:PMonfils@Lifespan.org] Sent: Friday, September 30, 2005 11:23 AM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Silicone slides He is probably referring to what we now call "Plus slides", which are coated with "Silane", a trade name for positively charged silicon tetrahydride. When these slides first came on the market some years ago, the name "Silane" was not yet in vogue, and the maufacturer described the coating on the slides simply as "electrostatically charged silicon". > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Carole Fields > Sent: Friday, September 30, 2005 8:05 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silicone slides > > Hi, > > I thought I had read something about silicone (coated) slides on the > Histo-Net but I may be mistaken. One of our pathologists asked if I I > knew > where to get silicone coated slides this morning? Have we discussed this > and I missed it? No idea yet why he wants this... > Thank you in advance for any help..... > > Carole Fields, HT,ASCP > Pathology Supervisor > Lexington Medical Center > 2720 Sunset Blvd. > W. Columbia, SC 29169 > > > > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From gcallis <@t> montana.edu Fri Sep 30 12:03:26 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 30 12:03:59 2005 Subject: [Histonet] fluorescent proteins (RFP and GFP) native fluorescence In-Reply-To: <61F5077F-B2B2-450C-AC41-3B4B0399703C@bidmc.harvard.edu> References: <61F5077F-B2B2-450C-AC41-3B4B0399703C@bidmc.harvard.edu> Message-ID: <6.0.0.22.1.20050930102022.01b1bf88@gemini.msu.montana.edu> Caroline, The first thing you need to do is go to Clontech website and access the Living Colours Manual for both GFP and DsRed. You can also talk to their technical service people when you have problems - they are excellent. We use a fusion protein with eGFP or DsRed is from Coral but the latter suffers from the same loss of fluorescence when fixed. We have found GFP, eGFP doesn't work well with fixation in general. I have not experienced either eGFP or DsRed photobleaching or failing to fluorescence (I am not sure what you mean by dispersion?) as long as I avoid using acetone or acetone/alcohol based fixatives. GFP, if you will, is as stated on title of that manual, best with living cells, although a fresh frozen sections, followed by a very short time in 1 -2 % paraformaldehyde often retains its fluorescence. By sliding microtome, do you mean a vibrating mictrome to cut into fresh tissue which can either be left unfixed or perfusion fixation of animal? Free floating sections should work just as well as a frozen section mounted on a slide. We now prefer to cryosection at 5 um (could go thicker if needed), mount UNFIXED sections on Erie Gold Plus Charge slides(undecalcified bone is sectioned with Cryojane tape transfer). An unfixed frozen section is dried at RT for a short time in dark. After air drying, the unfixed frozen sections are rinsed very gently with Dulbeccos PBS to get rid of OCT, then mounted with Prolong Gold Antifade Mounting Media, ready to use from Molecular Probes with DAPI - this is a hard set media. We seal the edges of coverslip with diluted permanent mounting media and NOT fingernail polish. Fingernail polish contains isopropyl alcohol that leaches into the PBS, causing the GFP to fade. Diluted mounting media contains either xylene or toluene, NOT miscible with aqueous PBS. You can purchase Prolong Gold with or without DAPI. DAPI gives us two color fluorescence with the DsRed fused protein where we need to see it on that on epithelial cell surfaces and blue nuclei with DAPI - a tidy way to get some sense of morphology. Unfixed tissues will have a bit of autoflourescence, and probably more with aldehyde induced fluorescence. You did not say if you were doing confocal laser scanning but if you are doing so, and use an inverted microscope, there are some delightful glass petri dishes with various size and thickness coverslip bottoms designed for inverted CLSM. Large, thick sections are far more easily viewed inverted with these dishes, and can be left free floating. We use these for vibratome cut thick 100 um sections stained with fluorophore conjugated antibodies (red) and allow the autofluorescence to be the counterfluorescence in yellow green colors. This type of viewing should work with GFP also. There is a confocal listserver you need to subscribe to out of Buffalo (Online search brought up University of Buffalo Confocal Listserver) which will help you out even more as there others doing the same thing you want to achieve. They can probably go into far more detail about GFP and all its chimeras, DsRed, and any instrumentation needed to view these bioluminescent proteins and specimen preparation. You can ask questions, search their archives, etc - it is valuable resource. Good luck, I am not sure I answered your questions but I strongly advise you to do to the confocal/Buffalo website. At 04:59 PM 9/29/2005, you wrote: >Hey guys, > >I was hoping someone here could give me some advice on their >experience with various fluorescent proteins. I need a good marker >for my viral vector. I have a humanized GFP, RFP and EGFP. I would >like to inject the vector into the tail vein to see which tissue >lights up. My hGFP variant is not very strong in vitro, but both RFP >and EGFP work well. I have heard that GFP quickly disperses when >mounted on slides. However, I have seen some papers that use native >EGFP fluorescence with cryosections and they don't have problems. >Ideally I would use floating tissue sections of 40 microns as I have >easy access to a sliding microtome. I have heard that RFP does not >work with fixation at all. > >I am open to any sort of advice. Could someone recommend a protocol >for visualizing native fluorescence with either EGFP or RFP? >Specifically, whether to fix or not, the thickness of the section, >floating sections vs. slide mounts, etc. > >Also, if there is another fluorescent protein available that you >could suggest I would like to hear about it. I have heard of yellow >and blue fluorescent proteins as well as a monomeric form or RFP. I >am also open to using a "universal" fusion protein, for example actin- GFP >if it solves the problem of GFP native fluorescence. > >Thanks, > >Caroline > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From sharon.osborn <@t> dnax.org Fri Sep 30 12:25:12 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 30 12:25:57 2005 Subject: [Histonet] RE: job postings for Palo Alto, CA Message-ID: <29B25753F6B1D51196110002A589D444023981F9@PALMSG30.us.schp.com> Histonetters! About a month ago I posted information that DNAX, ie., SP BioPharma would be having positions available for 2-4 people. We now have posted 4 positions looking for skill sets for defined areas. Job descriptions are posted on the Schering-Plough website. All the positions will have opportunity for cross-training. Please go to the main Schering-Plough web site at http://www.schering-plough.com/schering_plough/careers/careers.jsp (find our jobs by choosing CA Palo Alto as the location and Research and Development as the job category). When asked who referred you, please type in the name Jo Ann Katheiser so she receives credit for the referral. It is best to apply online for it seems to come through easier and quicker ac/c to channels. Questions? You may contact me directly, Sharon Osborn DNAX, SP BioPharma Palo Alto, CA 650.496.6539 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From jimr0712 <@t> comcast.net Fri Sep 30 12:50:07 2005 From: jimr0712 <@t> comcast.net (jimr0712@comcast.net) Date: Fri Sep 30 12:50:34 2005 Subject: [Histonet] Cost/test in anatomic pathology Message-ID: <093020051750.6039.433D7ACE000EC969000017972200745672CDCEC9CF9D030706@comcast.net> We are currently trying to verify costs/test in Anatomic Pathology. Is anyone aware of a good source(s) for this info ?? Thanks. -- Jim Robinson, M.S., HTL/HT (ASCP) Operations Manager Orizon Diagnostics, LLC 102 Chestnut Avenue Westmont, Illinois 60559 jimr0712@comcast.net 630-321-1506 (fax) 312-339-1169 (cell) From jamie.erickson <@t> abbott.com Fri Sep 30 13:08:12 2005 From: jamie.erickson <@t> abbott.com (Jamie E Erickson) Date: Fri Sep 30 13:08:35 2005 Subject: [Histonet] Re: fluorescent proteins (RFP and GFP) native fluorescence Message-ID: Caroline, I have had experience with looking at vectors in Mouse tissue years ago using EGFP as a report gene and had no trouble fixing/processing/embedding these samples in paraffin on my standard protocol. EGFP is very stable to these harsh conditions. I presented this data at the NSH poster session back in 99. Here was the conclusions, If you would like a copy of the methods or the poster contact me and I'll send you the Powerpoint poster. CONCLUSIONS The fluorescence of EGFP was still visible after routine fixation, processing, and paraffin embedding using ethanol and methanol as the dehydrating agent. For two of the three fixatives, EGFP fluorescence was still visible after being in fixative for as long as one month. A decrease in fluorescence was observed using the zinc-based fixative (Z-fix), compared to formalin or paraformaldehyde fixation. The fluorescence intensity decreased with increasing length of fixation. EGFP fluorescence appeared optimal using 10% NBF at the12 and 24 hour timepoints. Methanol processing appeared to better preserve the EGFP fluorescence using the zinc-based fixative (Z-fix),and at the later timepoints. These data suggest that routine histologic procedures and the use of common fixatives such as 10% neutral-buffered formalin can be used in the analysis of adenovirus-EGFP expression in vivo. Thus, the use of EGFP as a reporter molecule is an attractive alternative to -galactosidase for expression analysis. From PMonfils <@t> Lifespan.org Fri Sep 30 13:27:59 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 30 13:28:20 2005 Subject: [Histonet] VIR question - finfish tags Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175E9@lsexch.lsmaster.lifespan.org> Carol, Such a project depends largely on the chemical composition of the synthetic material. We section a variety of synthetics here since we are a core facility serving a large number of different researchers in various fields. Most synthetics can be sectioned in paraffin - once you get them into paraffin - but that can be tricky. In order to embed them in paraffin, you first have to make sure that the particular synthetic is chemically resistant to your dehydrant and your clearing agent, and is not harmed by the temperature of your processing paraffin. We routinely section sponge implants that have been removed from animals after several months duration. These are highly resistant chemically, and can be processed on a normal processing schedule using ethanol and xylene. We also work with resorbable vessel grafts, and these again can be processed normally. However, in another project we are sectioning bone containing a fine resorbable polymer screen, and xylene is not suitable for these. It makes the material soft and gummy. We found that tetrahydrofuran works well as a clearing agent for that project. More recently we have been doing work on an implantable device which contains synthetic tubules. Before we began work on this, I did what I always do in such cases - asked the researcher to send me samples of the synthetic material to test against various solvents before we started processing specimens. This one proved to be challenging. Xylene caused the material to shrink to a third its former size, and adjacent tubules to fuse into a solid mass. A few other solvents had a similar effect. Tetrahydrofuran, chloroform, toluene, and several other solvents dissolved the tubules completely within seconds. After trying over different 15 clearing agents, and rapidly running out of options, I tried carbon tetrachloride. It worked beautifully, rapidly clearing the tubules without causing any distortion at all. But carbon tetrachloride is very expensive and also very toxic, so I kept looking. Finally I discovered that a clearing agent called "SafeClear" by Fisher worked virtually as well as carbon tetrachloride, so that's what we are now using on this project. The blocks cut beautifully. Then of course we also have to use SafeClear to deparaffinize the slides before staining, and to clear them after staining. And, we had to find a coverslip mounting medium that is xylene and toluene free, and compatible with both the clearing agent and the synthetic material. Fortunately, VectaMount by Vector Laboratories proved suitable. So now we have our protocol in place for this specific project. One other consideration. Since most synthetics do not have a net negative charge like most tissues do, + charged slides do not hold them in place. Therefore you have to use a chemical adhesive of some sort. Depending on the particular synthetic, I usually use either PVA (polyvinyl acetate) or "Sta-On", a chrome gelatin product sold by Surgipath. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > McCollough, Carol > Sent: Friday, September 30, 2005 8:34 AM > To: Histonet (E-mail 2) > Subject: [Histonet] VIR question - finfish tags > > Greetings Histonetters: > > We have had a request from a researcher at Woods Hole for embedding and > sectioning of monkfish that have been tagged. The investigator is > interested in the interface between the living tissue and the tag. I > believe that these are Floy-type spaghetti tags, which are flexible soft > plastic, maybe latex. I have no experience with sectioning non-biological > materials. We do 99.999% paraffin here, but do have the capability to do > methacrylates and resins. Suggestions? > > Regards - > Carol > ********************** > Carol B. McCollough, HT/HTL(ASCP) > Aquatic Animal Research Pathologist > Oyster Disease Research Project > Fisheries Service > Maryland Department of Natural Resources > Cooperative Oxford Laboratory > 904 S. Morris Street > Oxford, Maryland 21654 > 410-226-5193 x124 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gcallis <@t> montana.edu Fri Sep 30 13:43:15 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 30 13:43:34 2005 Subject: [Histonet] CXCR4 and VEGF Receptors In-Reply-To: References: Message-ID: <6.0.0.22.1.20050930123703.01b57528@gemini.msu.montana.edu> R&D Systems has an absolutely spectacular photo of human astrocytes stained with mouse antiHuman CXCR4 monoclonal (Cat #MAB172) and antimouse IgG -Rhodamine conjugate secondary, and counterstained with DAPI in their 2005 desktop calendar (month of October). If it works for for IFA, it should work for IHC. Wen Sheng, Minneapolois Medical Research Foundation, Minneapolis did the staining. At 07:49 AM 9/30/2005, you wrote: >Hello All, > >I am just trying to get a broad feel for what antibodies people are using >lately for CXCR4 and VEGF receptors in both human and mouse tissue. > >Thanks, >Andrea >-- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From cgfields <@t> lexhealth.org Fri Sep 30 14:19:53 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 30 14:17:59 2005 Subject: [Histonet] Silicone slides Message-ID: That's exactly what I told him. He said no....Paul Monfils said he probably read something that said silicon tetrahydride which Silane slides are. I am going to ask him if that is what he read. THX Carole -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Friday, September 30, 2005 3:02 PM To: Carole Fields Subject: Re: [Histonet] Silicone slides I think you mean silane, not silicone, and the silane coated slides are Plus Charge from Erie. At 09:05 AM 9/30/2005, you wrote: >Hi, > >I thought I had read something about silicone (coated) slides on the >Histo-Net but I may be mistaken. One of our pathologists asked if I I knew >where to get silicone coated slides this morning? Have we discussed this >and I missed it? No idea yet why he wants this... >Thank you in advance for any help..... > >Carole Fields, HT,ASCP >Pathology Supervisor >Lexington Medical Center >2720 Sunset Blvd. >W. Columbia, SC 29169 > > > > >_________________________________ >This email and any files transmitted with it may contain PRIVILEGED or >CONFIDENTIAL information and may be read or used only by the intended >recipient. If you are not the intended recipient of the email or any of its >attachments, please be advised that you have received this email in error >and that any use, dissemination, distribution, forwarding, printing, or >copying of this email or any attached files is strictly prohibited. If you >have received this email in error, please immediately purge it and all >attachments and notify the sender by reply email or contact the sender at >the number listed above if one is provided. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From anh2006 <@t> med.cornell.edu Fri Sep 30 14:35:21 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 30 14:30:57 2005 Subject: [Histonet] CXCR4 and VEGF Receptors In-Reply-To: <6.0.0.22.1.20050930123703.01b57528@gemini.msu.montana.edu> References: <6.0.0.22.1.20050930123703.01b57528@gemini.msu.montana.edu> Message-ID: Thanks Gayle. We have been looking into this antibody based on some publications we have read so it's good to know someone has had good experiences with it. Does anyone know how to contact this person? I can't find information anywhere. Even searched the MMRF site. Thanks, Andrea At 12:43 PM -0600 9/30/05, Gayle Callis wrote: >R&D Systems has an absolutely spectacular photo of human astrocytes >stained with mouse antiHuman CXCR4 monoclonal (Cat #MAB172) and >antimouse IgG -Rhodamine conjugate secondary, and counterstained >with DAPI in their 2005 desktop calendar (month of October). If it >works for for IFA, it should work for IHC. Wen Sheng, Minneapolois >Medical Research Foundation, Minneapolis did the staining. -- From jkiernan <@t> uwo.ca Fri Sep 30 14:35:08 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 30 14:35:29 2005 Subject: [Histonet] Silicone slides References: Message-ID: <433D936C.AF538497@uwo.ca> Silicone must surely be an error for silanized slides. These are treated with aminotriethylaminosilane (APES), which combines chemically with the polysilicate ions of the glass. This leaves the surface covered with aminopropyl groups, which are protonated (positively charged) in the presence of slightly acidic water. See http://publish.uwo.ca/~jkiernan/adhesivs.htm for more information. It's easy to make your own silanized slides, or you can buy them ready-made; they have names like Plus. Silicone treatment of glass usually means reacting it with a hydrophobic silane derivative to make the surface non-wettable. This is the last thing you'd want for mounting sections. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Carole Fields wrote: > > Hi, > > I thought I had read something about silicone (coated) slides on the > Histo-Net but I may be mistaken. One of our pathologists asked if I I knew > where to get silicone coated slides this morning? Have we discussed this > and I missed it? No idea yet why he wants this... > Thank you in advance for any help..... > > Carole Fields, HT,ASCP > Pathology Supervisor > Lexington Medical Center > 2720 Sunset Blvd. > W. Columbia, SC 29169 > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rebecca.riesen <@t> dsilabs.com Fri Sep 30 16:33:11 2005 From: rebecca.riesen <@t> dsilabs.com (Riesen, Rebecca) Date: Fri Sep 30 16:33:36 2005 Subject: [Histonet] Cerner Millenium HELP Message-ID: <05844092236E7749A1BBBCB1077C34A2DEA0BA@dsi-ex01.gateway.dom> I have drawn the short straw and have the honor of being the AP representative for the migration to Cerner Millenium computer system for our hospitals. We currently have Cerner Classic. I was wondering if there are any HistoNet GURUS out there that have already gone through this possibly painful process and could share some of your wisdom and warnings with me. Any help is GREATLY appreciated. We start next month with our first big build. THANKS! DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From jkiernan <@t> uwo.ca Fri Sep 30 23:56:28 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 30 23:56:48 2005 Subject: [Histonet] gold chloride References: <6E1A8C242B4D6447A14A170D98A45C63EF18@barracuda.dcla.com> Message-ID: <433E16FC.6843AFDB@uwo.ca> Gold chloride (meaning tetrachloroauric acid or its sodium salt) does not need to be refrigerated and it is not light-sensitive. Aqueous solutions keep for years in clear glass bottles on an ordinary shelf. They slowly deteriorate with repeated use, becoming paler yellow with a subtle greyish sheen, and should then be recycled. The recycling method is simple and you recover about half the original gold chloride. For details, see Stain Technology 52:245-248 (1977). John Kiernan Anatomy, UWO London, Canada. ---------------------------------------- Jerry Duncan wrote: > > Fellow histologists, > > We currently prepare gold chloride rather than purchase the prepared liquid. > Does gold chloride need to be refrigerated after preparation? > > Thanks, > Jerry > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet