[Histonet] Re:xylene times

Carl Hobbs carl.hobbs <@t> kcl.ac.uk
Wed Jun 29 13:06:42 CDT 2005


Manual: 2x xylene 10mins each
             4x IMS 74OP ( Industrial methylated spirit) - 2mins each
              Rinse well in tap water. Proceed to technique
----- Original Message ----- 
From: <histonet-request <@t> lists.utsouthwestern.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, June 29, 2005 6:02 PM
Subject: Histonet Digest, Vol 19, Issue 42


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> Today's Topics:
>
>   1. Bowies method (Jose Luis Palazon Fernandez)
>   2. XYLENE EVAPORATION AND GLASS RACKS (Tim Wheelock)
>   3. Re: uneven haematoxylin/eosin staining (Janice A Mahoney)
>   4. Silly question about deparaffinization (Eva C Andersson)
>   5. RE: Silly question about deparaffinization (Sebree Linda A.)
>   6. Re: Silly question about deparaffinization (Pamela Marcum)
>   7. RE: Blocking serum (C.M. van der Loos)
>   8. RE: Silly question about deparaffinization (Rittman, Barry R)
>   9. RE: Phenol green gram stain (Monfils, Paul)
>  10. tungsten carbide knife resharpening (Nancy W. Troiano)
>  11. tungsten carbide resharpening (Nancy W. Troiano)
>  12. mouse ankles for IHC (Dana Marshall)
>  13. New Mexico Society for Histology (Breeden, Sara)
>  14. OCT (Lewis, Sarah)
>  15. detachment of tissue from OCT compound (Monfils, Paul)
>  16. Full time Histotech Opening (Nava, Josefa)
>  17. Pig Skin immunohistochemistry-losing sections during antigen
>      retrieval (SABYASACHI BISWAS)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Wed, 29 Jun 2005 15:00:41 +0200 (CEST)
> From: Jose Luis Palazon Fernandez <jluis.palazon <@t> icman.csic.es>
> Subject: [Histonet] Bowies method
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <20050629130041.C67348238AD <@t> perceval.uca.es>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Dear List-fellows
>
> Could any of you send me the protocol of Bowie´s method for 
> juxtaglomerular cells in kidney.
>
> thanks in advance
>
> José Luis
>
>
> Universidad de Oriente-Isla Margarita-Venezuela
> actualmente en: Instituto de Ciencias Marinas de Andalucia
> Puerto Real, Cádiz, España.
> email: jluis.palazon <@t> icman.csic.es
>
>
>
>
>
> ------------------------------
>
> Message: 2
> Date: Wed, 29 Jun 2005 09:42:27 -0400
> From: Tim Wheelock <twheelock <@t> mclean.harvard.edu>
> Subject: [Histonet] XYLENE EVAPORATION AND GLASS RACKS
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <42C2A543.8010400 <@t> mclean.harvard.edu>
> Content-Type: text/plain; charset=ISO-8859-1; format=flowed
>
> Hi Margaret:
>
> Perhaps the way  to stop xylene evaporation is to put something on top
> of the containers like a cork board, or heavy cookie sheet, anything
> heavy enough that will help seal the xylene in the containers.
>
> Speaking of glass racks, they do seem to be somewhat fragile. I tried
> them out in an attempt to batch my silver stains years ago  but they did
> not hold enough slides.
>
> I was wondering if anyone out there knew of glass racks that held 60
> slides, not back-to-back, but just one slide per slot.
> I am still trying to find a way to batch the Bielschowsky silver stains,
> instead of dealing with 14 different coplan jars.
>
> Tim Wheelock
> Harvard Brain Bank
> McLean Hospital
>
>
>
> ------------------------------
>
> Message: 3
> Date: Wed, 29 Jun 2005 08:57:14 -0500
> From: "Janice A Mahoney" <jmahoney <@t> alegent.org>
> Subject: Re: [Histonet] uneven haematoxylin/eosin staining
> To: <histonet <@t> pathology.swmed.edu>
> Message-ID: <s2c26273.048 <@t> mail.alegent.org>
> Content-Type: text/plain; charset=US-ASCII
>
> I have experienced this problem at several labs where I have worked. 
> Finally figured out it happens more in the summer when the humidity is 
> high.  It was due to incomplete removal of paraffin due to moisture in the 
> xylene.  Try changing it more often or filtering it.  The filter paper 
> will labsorbe the water.
> Jan
> Omaha NE
>
>>>> Paul Bradbury <histology.bc <@t> shaw.ca> 06/28/2005 6:23:27 PM >>>
> Sounds like a classical case of incomplete removal of wax before
> staining begins. Check the de-waxing reagents and times.
>
> Paul Bradbury
> Kamloops, BC
> Canada
>
>
> Diana McCaig wrote:
>
>>We have been experiencing uneven staining recently.  Even when all 
>>sections
>>are cut on the same microtome and put in the same rack on an autostainer,
>>their macroscopic appearance show incredible variations.  It is so random 
>>we
>>can not identify the problem.  The Harris hemotoxylin is filtered daily.
>>The tissue type does not make any difference.  We can have 4 prostate 
>>slides
>>and 2 will be good and 2 will show random staining intensities.  Recuts 
>>may
>>show variation in a different area which makes me think it is the staining
>>and not the cutting.
>>
>>Any suggestions would be truly appreciated.
>>Diana McCaig, MLT
>>
>>
>>_______________________________________________
>>Histonet mailing list
>>Histonet <@t> lists.utsouthwestern.edu
>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>>
>>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
>
>
> ------------------------------
>
> Message: 4
> Date: Wed, 29 Jun 2005 09:56:36 -0400
> From: Eva C Andersson <eca9 <@t> georgetown.edu>
> Subject: [Histonet] Silly question about deparaffinization
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <42C2A894.7070708 <@t> georgetown.edu>
> Content-Type: text/plain; charset=us-ascii; format=flowed
>
> Good morning,
> I am probably asking a very silly question but I recently spoke to
> someone about deparaffinization and hydration. I found that the times
> that they use and the times that we use are very different. So my
> question to you is this: What lenghts of time do you use for Xylenes,
> 100% alcohol, 95% alcohol and 70% alcohol?
> It is about deparaffinising charged (+) slides. So nothing fancy.
> Just curious. Maybe I can cut back some on the timed that we use which 
> are:
> 30min Xylenes, 10min 100%, 10min 95% and 6min 70%.
> Thanks for your responses.
> Eva Andersson
> Georgetown University
>
>
>
> ------------------------------
>
> Message: 5
> Date: Wed, 29 Jun 2005 09:13:44 -0500
> From: "Sebree Linda A." <la.sebree <@t> hosp.wisc.edu>
> Subject: RE: [Histonet] Silly question about deparaffinization
> To: "Eva C Andersson" <eca9 <@t> georgetown.edu>,
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <D6B654003615874B873E15BA680E2D2211352BFD <@t> uwhis-xchng1.hosp.wisc.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> We deparaffinize in 3 changes of xylene, 2 changes of 100% ETOH, 1 95%
> ETOH and 1 70% ETOH, all for 3 minutes each.  We never skimp on the
> xylene times but have been known to shorten the ETOH times if we
> manually agitate the slides and are in a real hurry.
>
> 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 <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Eva C
> Andersson
> Sent: Wednesday, June 29, 2005 8:57 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Silly question about deparaffinization
>
>
> Good morning,
> I am probably asking a very silly question but I recently spoke to
> someone about deparaffinization and hydration. I found that the times
> that they use and the times that we use are very different. So my
> question to you is this: What lenghts of time do you use for Xylenes,
> 100% alcohol, 95% alcohol and 70% alcohol?
> It is about deparaffinising charged (+) slides. So nothing fancy. Just
> curious. Maybe I can cut back some on the timed that we use which are:
> 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your
> responses. Eva Andersson Georgetown University
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 6
> Date: Wed, 29 Jun 2005 10:21:59 -0400
> From: Pamela Marcum <pmarcum <@t> vet.upenn.edu>
> Subject: Re: [Histonet] Silly question about deparaffinization
> To: Eva C Andersson <eca9 <@t> georgetown.edu>,
> histonet <@t> lists.utsouthwestern.edu
> Message-ID: <6.1.1.1.2.20050629101442.019659d8 <@t> mail.vet.upenn.edu>
> Content-Type: text/plain; charset="us-ascii"; format=flowed
>
> Eva,
>
> Depending on the thickness of the sections my times can vary. Sections of 
> 4
> to 6 micron are deparaffinized in our laboratory at 2 changes at 5 minutes
> each with alcohols at 3 minutes each per station to water.  Sections
> thicker than 6 microns may go as long 15 minutes X 2 in xylenes and 5 to 
> 10
> minutes in each alcohol.  I am very paranoid about getting all of the
> paraffin (or MMA where I use one hour per change for xylenes) so I may go
> over board.  I also change my reagents more often in humid weather.  I do
> not use an automated stainer so agitation only takes place at the 
> beginning
> of the changes to assure the last reagent is rinsed off.
>
> Hope this helps.
>
> At 09:56 AM 6/29/2005, Eva C Andersson wrote:
>>Good morning,
>>I am probably asking a very silly question but I recently spoke to someone
>>about deparaffinization and hydration. I found that the times that they
>>use and the times that we use are very different. So my question to you is
>>this: What lenghts of time do you use for Xylenes, 100% alcohol, 95%
>>alcohol and 70% alcohol?
>>It is about deparaffinising charged (+) slides. So nothing fancy.
>>Just curious. Maybe I can cut back some on the timed that we use which 
>>are:
>>30min Xylenes, 10min 100%, 10min 95% and 6min 70%.
>>Thanks for your responses.
>>Eva Andersson
>>Georgetown University
>>
>>_______________________________________________
>>Histonet mailing list
>>Histonet <@t> 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 <@t> vet.upenn.edu
>
>
>
>
>
> ------------------------------
>
> Message: 7
> Date: Wed, 29 Jun 2005 16:29:34 +0200
> From: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>
> Subject: [Histonet] RE: Blocking serum
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <10b1c3d10b6fff.10b6fff10b1c3d <@t> amc.uva.nl>
> Content-Type: text/plain; charset="us-ascii"
>
>
>   Dear Baowei Peng,
>
>   Since  you  have  a  goat  anti-rabbit  IgG as second step reagent you
>   should  use  normal  goat  serum  for pre-blocking step. Otherwise use
>   Protein Block, serum-free (DakoCyto X0909).
>
>   Chris van der Loos, PhD
>   Dept. of Pathology
>   Academical Medical Center M2-230
>   Meibergdreef 9
>   NL-1105 AZ Amsterdam
>   The Netherlands
>
>   phone:  +31 20 5665631
>   fax:    +31 20 6960389
>
>   Date: Wed, 29 Jun 2005 08:35:04 +0800 (BEIST)
>   From: Baowei Peng <pengbw <@t> sjtu.edu.cn>
>   Subject: [Histonet] Blocking serum
>   To: histonet <@t> lists.utsouthwestern.edu
>   Message-ID: <20050629003504.7EDCD11104FD <@t> sjtu.edu.cn>
>   Content-Type: text/plain; charset="gb2312"
>   Hello, all
>   I\'m doing IHC on mouse frozen sections. The first antibody in my hand
>   is  Rabbit polyclonal antibody,the second antibody is Goat anti Rabbit
>   IgG(H+L).  I\'m wondering what kind of serum I should use to block non
>   specific staining.
>   Thanks in advance!
>   Baowei Peng
>   1954 Huashan Road
>   Pharmacy School
>   Shanghai Jiaotong University
>   Shanghai, 200030
>   China
>   Ph:86-21-62932108
>   E-mail:pengbw <@t> sjtu.edu.cn
>
>
> ------------------------------
>
> Message: 8
> Date: Wed, 29 Jun 2005 09:31:25 -0500
> From: "Rittman, Barry R" <Barry.R.Rittman <@t> uth.tmc.edu>
> Subject: RE: [Histonet] Silly question about deparaffinization
> To: "histonet" <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <EA1FDD2A141B7448B4B1AFFFCAC08DE403AB26F6 <@t> UTHEVS1.mail.uthouston.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> Eva
> I would suggest that you do a trial and error run.
> Deparaffinization time with xylene will depend on several factors
> including: thickness of sections, type of paraffin, number of slides,
> agitation, temperature of solution, volume of fluid, number of times
> that xylene bath was used, quality of the xylenes solution and so on.
> I have always erred on the conservative side and given a longer time as
> you have done in the past. However, for many labs, as Linda has
> indicated, 3 minutes in each bath is fine.
> Suggest that you try a set of slides and deparaffinize them for 3, 5 7
> and 10 minutes and compare your results.
> Barry
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sebree
> Linda A.
> Sent: Wednesday, June 29, 2005 9:14 AM
> To: Eva C Andersson; histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] Silly question about deparaffinization
>
> We deparaffinize in 3 changes of xylene, 2 changes of 100% ETOH, 1 95%
> ETOH and 1 70% ETOH, all for 3 minutes each.  We never skimp on the
> xylene times but have been known to shorten the ETOH times if we
> manually agitate the slides and are in a real hurry.
>
> 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 <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Eva C
> Andersson
> Sent: Wednesday, June 29, 2005 8:57 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Silly question about deparaffinization
>
>
> Good morning,
> I am probably asking a very silly question but I recently spoke to
> someone about deparaffinization and hydration. I found that the times
> that they use and the times that we use are very different. So my
> question to you is this: What lenghts of time do you use for Xylenes,
> 100% alcohol, 95% alcohol and 70% alcohol?
> It is about deparaffinising charged (+) slides. So nothing fancy. Just
> curious. Maybe I can cut back some on the timed that we use which are:
> 30min Xylenes, 10min 100%, 10min 95% and 6min 70%. Thanks for your
> responses. Eva Andersson Georgetown University
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 9
> Date: Wed, 29 Jun 2005 10:32:07 -0400
> From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
> Subject: RE: [Histonet] Phenol green gram stain
> To: "'histonet <@t> lists.utsouthwestern.edu'"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <09C945920A6B654199F7A58A1D7D1FDE0171755E <@t> lsexch.lsmaster.lifespan.org>
>
> Content-Type: text/plain; charset="ISO-8859-1"
>
> I'm not sure what "phenol green" is?  But the Gram-Twort technique is a 
> gram
> stain with a green background, due to Fast Green FCF added to the red 
> stain.
> Perhaps that's what you are looking for. You can find that technique here:
>
> http://www.scienceboard.net/resources/protocols.asp?action=article&protocol_
> id=310&criteria=
>
>
>
> ------------------------------
>
> Message: 10
> Date: Wed, 29 Jun 2005 10:55:20 -0400
> From: "Nancy W. Troiano" <nancy.troiano <@t> yale.edu>
> Subject: [Histonet] tungsten carbide knife resharpening
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <5.2.1.1.2.20050629105118.00c05630 <@t> email.med.yale.edu>
> Content-Type: text/plain; format=flowed; charset=us-ascii
>
> We have our tungsten carbide knives sharpened by Dorn/Hart Microedge, Inc.
> 135 Home Avenue Villa Park, IL 60181.   Telephone number is
> 630-832-3843.  They are great - quality work at a reasonable price, quick
> turnaround time.  I have sent my tungsten carbide knives to them for the
> last 25 years for resharpening!
>
> Nancy Troiano, M.S., Associate in Research III
> Yale Core Center for Musculoskeletal Disorders
> Yale University
> P.O. Box 208071
> New Haven, CT  06520
> (203)785-5136
>
>
>
>
> ------------------------------
>
> Message: 11
> Date: Wed, 29 Jun 2005 10:57:31 -0400
> From: "Nancy W. Troiano" <nancy.troiano <@t> yale.edu>
> Subject: [Histonet] tungsten carbide resharpening
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <5.2.1.1.2.20050629105626.00c11870 <@t> email.med.yale.edu>
> Content-Type: text/plain; format=flowed; charset=us-ascii
>
> Forgot to mention that the price to resharpen a knife from Dorn/Hart is
> $150.00 for a 16 cm  tungsten carbide knife.  Turnaround time is
> approximately one week.
>
>
>
>
> ------------------------------
>
> Message: 12
> Date: Wed, 29 Jun 2005 10:24:23 -0500
> From: "Dana Marshall" <dmarsha3 <@t> utmem.edu>
> Subject: [Histonet] mouse ankles for IHC
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000a01c57cbe$a0d08de0$f623c084 <@t> DanaM>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi everyone,
>
> I am gearing up for mouse ankle-joint histology.  (I may do vertebral 
> column eventually as well if anyone has done that.)  I have a Cryojane 
> tape transfer system and have done a bit of sectioning of the 
> undecalcified bones and they look adequate.  I haven't done any fancy 
> staining yet.  My question is a fairly broad one in that I wanted to ask 
> those who might be doing this for a brief outline of their procedure from 
> beginning to end.  So far I am fixing in 10% formalin (in H2O) for a 
> couple of days.  I embed the joint with the heel towards the bottom of the 
> block (I thought it might be more stable to cut up from the heel but 
> perhaps not?)  Ultimately I will do IHC staining and, possibly, some in 
> situ hybridization.  I have some previous info in regards to knives and 
> angles of cutting etc and would appreciate any additional insights into 
> that as well.
>
> I am having trouble stringing together the information that will allow me 
> to make wise choices early on to maximize the use of my samples (and my 
> mice).  Any advice would be GREATLY appreciated.
>
> Thanks again.
>
> Dana (who dabbles in histology every 10 years whether I need to or not:-)
>
>
>
> ------------------------------
>
> Message: 13
> Date: Wed, 29 Jun 2005 09:29:01 -0600
> From: "Breeden, Sara" <sbreeden <@t> nmda.nmsu.edu>
> Subject: [Histonet] New Mexico Society for Histology
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <F16987B849BBD649AA3CD478B07F81F710528F <@t> nmdamail.nmda.ad.nmsu.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> The New Mexico Society for Histology, after having been rather inactive
> for the past couple years, is preparing to go "back on track"!  If you
> are a histologist (or have an interest therein) and live in New Mexico
> or one of our adjoining states, please consider joining us.  If you have
> been a member and would like to reactivate, we'd love to have you back.
> There will be a general business meeting and short presentation on
> Saturday, August 6th, beginning at 10:00 a.m.; the meeting will be held
> at Tricore Reference Laboratories, 1001 Woodward Place NE, Albuquerque.
> If interested in either the meeting, or membership, please contact me at
> this email address, or at NMHISTO <@t> AOL.COM.
>
> Sally Breeden, HT(ASCP)
> New Mexico Department of Agriculture
> Veterinary Diagnostic Services
> POBox 4700
> Albuquerque, NM  87196
> (505)841-2576
> (505)841-2518 FAX
>
>
>
> ------------------------------
>
> Message: 14
> Date: Tue, 28 Jun 2005 16:03:42 -0400
> From: "Lewis, Sarah" <LewisS <@t> pediatrics.ohio-state.edu>
> Subject: [Histonet] OCT
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <714B9F12B4E18C4C843B66E8E190F2AD447526 <@t> res2k3ms01.CRII.ORG>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hello!
>
> I use 7% gum Tragacanth that I make up myself using 100ml of distilled 
> water, 7gms of gum Tragacanth, and 5 grains of Thymol. Dissolve by 
> manually stirring at rm temp.(may take several hours) Store half 
> refrigerated(4c) and half frozen
> (-20c). I work with only muscle and nerve biopsy tissue but this works 
> great for me. I am sure this is a bit "old school", but I have never had 
> any trouble with cracking or stability of the tissue. It also seems to be 
> great for long term storage in -80 freezer. Good luck!!
>
>
> Sarah E. Lewis
> Histotechnician
> Childrens Research
> Center for Gene Therapy
> 700 Childrens Dr Rm WA3112
> Columbus OH 43205
> (614)-722-2204
> LewisS <@t> ccri.net
>
>
>
> -----Original Message----- 
> From: ssq5977 <@t> yahoo.com.cn [mailto:ssq5977 <@t> yahoo.com.cn]
> Sent: Thursday, June 16, 2005 9:30 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Is there anything better than OCT?
>
>
>
> Hi all:
>
> We are currently using OCT to embed renal biopsy tissue in cryostat mold. 
> First of all ,we put a drop of OCT on the mold ,then we put the tissue on 
> the top of the OCT and dip the mold into liquid nitrogen for 10 seconds. 
> then the tissue is sent to cryosection. Does anyone know if there is 
> anything better than OCT to embed the tissue?
>
> Thanks!
>
> Shuqiong Shen(ssq5977 <@t> yahoo.com.cn)
> Research Institute of Nephrology, Jinling Hospital
> 305 East Zhongshan Road, Nanjing
> Jiangsu Province,P.R of China
>
> Zip code: 210002
> Tel: +86 25 85912177
>
>
>
>
> Sarah E. Lewis
> Histotechnician
> Childrens Research
> Center for Gene Therapy
> 700 Childrens Dr Rm WA3112
> Columbus OH 43205
> (614)-722-2204
> LewisS <@t> ccri.net
>
>
>
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> ------------------------------
>
> Message: 15
> Date: Wed, 29 Jun 2005 11:35:51 -0400
> From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
> Subject: [Histonet] detachment of tissue from OCT compound
> To: "'histonet <@t> lists.utsouthwestern.edu'"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <09C945920A6B654199F7A58A1D7D1FDE0171755F <@t> lsexch.lsmaster.lifespan.org>
>
> Content-Type: text/plain; charset="ISO-8859-1"
>
> Someone has sent me 60 rat brains frozen in OCT compound (whole brain 
> except
> for the cerebellum and the anterior 1 cm). He wants multiple 30-micron
> sections from each brain. They appear to be well infiltrated with sucrose,
> and they cut smoothly on the cryostat at -20C. However, the sections of
> tissue fall out of the surrounding OCT, which makes it very difficult to
> pick them up on slides without a lot of wrinkles and bubbles.
>
> Any tricks to deal with this situation?
>
> Paul M.
>
>
>
> ------------------------------
>
> Message: 16
> Date: Wed, 29 Jun 2005 10:56:47 -0500
> From: "Nava, Josefa" <JosefaNava <@t> texashealth.org>
> Subject: [Histonet] Full time Histotech Opening
> To: <histonet <@t> pathology.swmed.edu>
> Message-ID:
> <2C515C1049EAF5459EFD8C9B929078A41944B6 <@t> phdex03.txhealth.org>
> Content-Type: text/plain; charset="us-ascii"
>
> We are looking for a full time Histotech or  ASCP  eligible position  at
> Presbyterian Hospital of Dallas, Texas. Interested candidates may
> contact Maureen Hoops at 214-345-7795 for details and benefit
> information.
>
>
>
>
>
>
>
>
>
> 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.
>
> ------------------------------
>
> Message: 17
> Date: Wed, 29 Jun 2005 12:33:50 -0400
> From: SABYASACHI BISWAS <biswas.16 <@t> osu.edu>
> Subject: [Histonet] Pig Skin immunohistochemistry-losing sections
> during antigen retrieval
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <23a1309239d9f7.239d9f723a1309 <@t> osu.edu>
> Content-Type: text/plain; charset=us-ascii
>
> Hi All
> We are facing a problem with our pig skin biopsy samples. We are involved 
> in wound healing research and therefore have to do histological staining 
> of pig skin. Our biopsies are taken with 6mm biopsy punches around a 3mm 
> biopsy punch taken previously (the wound). The samples are fixed in 
> formalin and embedded in paraffin. We were losing sections even during H&E 
> staining but managed to solve that by baking the slides at 60degrees C for 
> 1hour instead of 37 degrees overnight; we also use Superfrost++ slides. 
> However during antigen retrieval using the microwave method we are losing 
> sections wholly or partially. Does anyone have experience with antigen 
> retrieval in pig skin samples. We appreciate any help/advice that you can 
> give us.
>
> Thanks
>
> Sabya Biswas
> The Ohio State University
>
>
>
>
> ------------------------------
>
> _______________________________________________
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> Histonet <@t> lists.utsouthwestern.edu
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>
> End of Histonet Digest, Vol 19, Issue 42
> ****************************************
>
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