[Histonet] RE: Histonet Digest, Vol 97, Issue 31
Goodwin, Diana
dgoodwin <@t> rwjuhh.edu
Wed Dec 28 12:53:11 CST 2011
RE: standard for sm bx microtomy (Sharon Allen)
Hi, Sharon.
We are cutting 2 H&E levels per block, with 1 extra slide and 1 special stain slide in between - the special being dependent on the type of tissue-HP Giemsa for upper GI, PAS/AB for esophagus, Mucin for lung, etc.
Hope this helps.
Diana G. Goodwin, BS, HT(ASCP)QIHC
Department of Pathology
Robert Wood Johnson University Hospital at Hamilton
One Hamilton Health Place
Hamilton, NJ 08690
Ph: 609.631.6996
Email: dgoodwin <@t> rwjuhh.edu
-----Original Message-----
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Sent: Wednesday, December 28, 2011 1:04 PM
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Subject: Histonet Digest, Vol 97, Issue 31
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Today's Topics:
1. Re: Histonet Digest, Vol 97, Issue 30 (Thomas "Tom" Ehlers)
2. Re: immuno stainers (Blazek, Linda)
3. good protocol for ICC in neurons (Guillermo Palchik)
4. Re: immuno stainers (Karen Lahti)
5. gel pad for microtome vibration (Nancy Schmitt)
6. FW: Quality Modules (Sharon Allen)
7. FW: standard for sm bx microtomy (Sharon Allen)
----------------------------------------------------------------------
Message: 1
Date: Tue, 27 Dec 2011 14:56:54 -0500
From: "Thomas \"Tom\" Ehlers" <tehlers818 <@t> gmail.com>
Subject: [Histonet] Re: Histonet Digest, Vol 97, Issue 30
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
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Hi Cindy,
What's the name of the new Dako stainer that you demo'd?
On Tue, Dec 27, 2011 at 1:00 PM,
<histonet-request <@t> lists.utsouthwestern.edu>wrote:
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> You can reach the person managing the list at
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> When replying, please edit your Subject line so it is more specific
> than "Re: Contents of Histonet digest..."
>
>
> Today's Topics:
>
> 1. RE: HSV1 and HSV2 (Hoekert, W.E.J.)
> 2. RELIA HOT Histology Job Alert. 12/27/2011 Great Opportunity
> for a histology tech in Virginia. (Pam Barker)
> 3. immuno stainers (Cynthia Pyse)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Tue, 27 Dec 2011 10:38:20 +0100
> From: "Hoekert, W.E.J." <W.E.J.Hoekert <@t> olvg.nl>
> Subject: RE: [Histonet] HSV1 and HSV2
> To: "Evans, Andria B" <aevans3 <@t> lghealth.org>,
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <1190CB05C44B13409483514729C2FC3601F84207 <@t> PAIT42.olvg.nl>
> Content-Type: text/plain; charset="iso-8859-1"
>
> For the immunologic pAB cocktail we are using Protease 1 (4 min) and ab
> incubation of 32 min. The only problem is that mast cells are also
> staining, it is a side effect of the protease 1.
>
> (AB concentration 1:400)
>
> Willem Hoekert
> OLVG Amsterdam
> The Netherlands
>
>
>
>
> ________________________________
>
> Van: histonet-bounces <@t> lists.utsouthwestern.edu namens Evans, Andria B
> Verzonden: vr 23-12-2011 18:48
> Aan: histonet <@t> lists.utsouthwestern.edu
> Onderwerp: [Histonet] HSV1 and HSV2
>
>
>
> I am currently having issues with our HSV1 and HSV2 staining tissue
> components that it shouldn't be/background. We are running Ventana
> Benchmark XTs and Ultras. Using Dako's polyclonal rabbit concentrate.
> Does anyone out there in histoland have a concentration with a protocol
> that works and looks clean?
>
> Thank you for your help!
>
> Andria B Evans HTL(ASCP)CM
> This email was sent securely from the LGHealth Email Service
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>
> ------------------------------
>
> Message: 2
> Date: Tue, 27 Dec 2011 11:26:52 -0500
> From: "Pam Barker" <relia1 <@t> earthlink.net>
> Subject: [Histonet] RELIA HOT Histology Job Alert. 12/27/2011 Great
> Opportunity for a histology tech in Virginia.
> To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <2AD338F5D60F42969C3852F762D09410 <@t> ownerf1abaad51>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi Histonetters,
> Seasons Greetings!!
> I hope everyone enjoyed a wonderful holiday weekend. I have a brand new
> histology job that I am pretty excited about and I wanted to take a
> minute and share the info with you:
>
> RELIA Solutions the nation?s only recruiting firm dedicated to the
> nationwide permanent placement of histology professionals is assisting a
> leading lab in Roanoke, VA in their search for a histology tech. ASCP
> HT/HTL and 3 years experience preferred. IHC is a plus. This is a day
> shift (early morning) M-F full time permanent position. My client
> offers a great salary and excellent benefits. For more information
> please contact Pam Barker at <mailto:relia1 <@t> earthlink.net>
> relia1 <@t> earthlink.net or toll free at 866-607-3542
>
> If you or anyone you know might be interested in hearing more about this
> opportunity please contact me. I can be reached at
> <mailto:relia1 <@t> earthlink.net> relia1 <@t> earthlink.net or toll free at
> 866-607-3542.
>
> Thanks-Pam
>
>
> 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: <mailto:relia1 <@t> earthlink.net> relia1 <@t> earthlink.net
> <http://www.facebook.comPamBarkerRELIA> www.facebook.comPamBarkerRELIA
> <http://www.linkedin.com/reliasolutions>
> www.linkedin.com/reliasolutions
> <http://www.myspace.com/pamatrelia> www.myspace.com/pamatrelia
> <http://www.twitter.com/pamatrelia> www.twitter.com/pamatrelia
>
>
>
> ------------------------------
>
> Message: 3
> Date: Tue, 27 Dec 2011 11:49:50 -0500
> From: "Cynthia Pyse" <cpyse <@t> x-celllab.com>
> Subject: [Histonet] immuno stainers
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <003e01ccc4b7$8cb51a40$a61f4ec0$@com>
> Content-Type: text/plain; charset="us-ascii"
>
> Hello Histonetters
>
> Happy Tuesday. I hope everyone had a Merry Christmas. It is amazing how
> times flies when you have time off. Now it's back to the old laboratory
> grind.
>
> We are in the market for a new immuno stainer. I currently use a Dako Link,
> which I am very pleased with. I have demoed the new Dako with great
> results,
> unfortunately we need 2 stainers and the pricing maybe a little out of our
> range.
>
> Is anyone using the Biocare Intellipath? What are the pro's and con's? How
> is their service? I currently use their detection system, Mach 4, but the
> majority of my antibodies are purchased from Dako. Any input on the daily
> use of the Intellipath from current users would be appreciated.
>
> Thanks in advance for the input
>
> Cindy
>
>
>
> Cindy Pyse, CLT, HT (ASCP)
>
> Laboratory Manager
>
> X-Cell Laboratories
>
> e-mail cpyse <@t> x-celllab.com
>
>
>
>
>
> ------------------------------
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> End of Histonet Digest, Vol 97, Issue 30
> ****************************************
>
------------------------------
Message: 2
Date: Tue, 27 Dec 2011 14:58:46 -0500
From: "Blazek, Linda" <lblazek <@t> digestivespecialists.com>
Subject: Re: [Histonet] immuno stainers
To: Cynthia Pyse <cpyse <@t> x-celllab.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<F7B33547-74AA-4071-B193-3C222C05A6DA <@t> digestivespecialists.com>
Content-Type: text/plain; charset="us-ascii"
I have the Intellipath from BioCare. I love it. Service is spectacular. It's very user friendly. You do have to be consistent with your cleaning protocol. In the past 3-4 years I have only had 1 day of down time. The company is very proactive when it comes to service. You couldn't ask for better a better company to work with in both sales or service. If you want to contact me directly feel.
Linda Blazek
GI Pathology
937 396-2623
Sent from my iPhone
On Dec 27, 2011, at 11:51 AM, "Cynthia Pyse" <cpyse <@t> x-celllab.com> wrote:
> Hello Histonetters
>
> Happy Tuesday. I hope everyone had a Merry Christmas. It is amazing how
> times flies when you have time off. Now it's back to the old laboratory
> grind.
>
> We are in the market for a new immuno stainer. I currently use a Dako Link,
> which I am very pleased with. I have demoed the new Dako with great results,
> unfortunately we need 2 stainers and the pricing maybe a little out of our
> range.
>
> Is anyone using the Biocare Intellipath? What are the pro's and con's? How
> is their service? I currently use their detection system, Mach 4, but the
> majority of my antibodies are purchased from Dako. Any input on the daily
> use of the Intellipath from current users would be appreciated.
>
> Thanks in advance for the input
>
> Cindy
>
>
>
> Cindy Pyse, CLT, HT (ASCP)
>
> Laboratory Manager
>
> X-Cell Laboratories
>
> e-mail cpyse <@t> x-celllab.com
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 3
Date: Tue, 27 Dec 2011 16:32:06 -0500
From: Guillermo Palchik <gp62 <@t> georgetown.edu>
Subject: [Histonet] good protocol for ICC in neurons
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <80DAFD47-401A-4FAF-AB05-32EC6AA81364 <@t> georgetown.edu>
Content-Type: text/plain; charset=us-ascii
Dear histonetters,
I am trying to do some ICC on nuclei of rat primary neurons that have been grown for 2 weeks on coverslips (seeded at 75,000 cells/ml). So far I have followed the protocol that a (long since gone) student in the lab used, but I have been unsuccessful. My nuclei end up looking like raisins....
The protocol that the student used is unlike anything I have seen:
Wash with cold PBS 2X 5 min
Rinse briefly with cold methanol
Fix in methanol at -20C for 20 minutes
Wash in cold PBS 2 X 10 min
Block in 5% NGS / 0.05% Triton-X for 6 hours at 4C
Add Primary antibodies diluted in Blocking Solution overnight @ 4C
Wash in cold Wash Solution (2% NGS/0.05% Triton-X) 3 X 10 min
Add Secondary Antibodies diluted in Blocking Solution for 1.5 hours at RT in dark
Wash in Wash Solution 3 X 20 min in dark
Wash in PBS 3 X 10 min in dark
Add DAPI (1:10,000 in PBS) for 5 minutes in dark
Wash in ddH2O 3 X 10 min in dark
Mount with Fluoro-Gel with Tris
Store @ 4C in dark until dry
First, after a methanol fixation of 20 minutes at -20C the protocol calls for blocking and permeabilizing the cells for 6 hours!
Second, it keeps washing with this washing solution that contains triton.
As I said, I have followed this protocol to the T and I keep getting horrible looking, shriveled cells and nuclei...
The question I have is, does the protocol look okay and maybe it is me, or should I change specific parameters of it?
Does anybody have a good working protocol that could share with me? It would be greatly appreciated...
Thanks
Gil
--
Guillermo Palchik
Ph.D. Candidate - Interdisciplinary Program in Neuroscience
Georgetown University Medical Center
Research Building Room W 217
3970 Reservoir Rd. NW, Washington, DC 20007
Lab: 202-687-7825
------------------------------
Message: 4
Date: Tue, 27 Dec 2011 19:34:36 -0700
From: Karen Lahti <karen <@t> gateslinger.com>
Subject: Re: [Histonet] immuno stainers
To: "Blazek, Linda" <lblazek <@t> digestivespecialists.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BEBB5A0E-E0AF-417B-8632-01E5CFF97858 <@t> gateslinger.com>
Content-Type: text/plain; charset=us-ascii
We have the IntelliPath as well. I have the same sentiments as Linda regarding all aspects of Biocare and the instrument. We use their antibodies and detection. The company as a whole is excellent to deal with and very customer oriented.
Thanks, Karen Lahti
Arizona Digestive Health
Phoenix, AZ
On Dec 27, 2011, at 12:58 PM, "Blazek, Linda" <lblazek <@t> digestivespecialists.com> wrote:
> I have the Intellipath from BioCare. I love it. Service is spectacular. It's very user friendly. You do have to be consistent with your cleaning protocol. In the past 3-4 years I have only had 1 day of down time. The company is very proactive when it comes to service. You couldn't ask for better a better company to work with in both sales or service. If you want to contact me directly feel.
> Linda Blazek
> GI Pathology
> 937 396-2623
>
> Sent from my iPhone
>
> On Dec 27, 2011, at 11:51 AM, "Cynthia Pyse" <cpyse <@t> x-celllab.com> wrote:
>
>> Hello Histonetters
>>
>> Happy Tuesday. I hope everyone had a Merry Christmas. It is amazing how
>> times flies when you have time off. Now it's back to the old laboratory
>> grind.
>>
>> We are in the market for a new immuno stainer. I currently use a Dako Link,
>> which I am very pleased with. I have demoed the new Dako with great results,
>> unfortunately we need 2 stainers and the pricing maybe a little out of our
>> range.
>>
>> Is anyone using the Biocare Intellipath? What are the pro's and con's? How
>> is their service? I currently use their detection system, Mach 4, but the
>> majority of my antibodies are purchased from Dako. Any input on the daily
>> use of the Intellipath from current users would be appreciated.
>>
>> Thanks in advance for the input
>>
>> Cindy
>>
>>
>>
>> Cindy Pyse, CLT, HT (ASCP)
>>
>> Laboratory Manager
>>
>> X-Cell Laboratories
>>
>> e-mail cpyse <@t> x-celllab.com
>>
>>
>>
>> _______________________________________________
>> 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: 5
Date: Wed, 28 Dec 2011 14:21:44 +0000
From: Nancy Schmitt <Nancy_Schmitt <@t> pa-ucl.com>
Subject: [Histonet] gel pad for microtome vibration
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<906B4DA90ED1DB4DB6C7E94D7CEE6C367BC785 <@t> PEITHA.wad.pa-ucl.com>
Content-Type: text/plain; charset="us-ascii"
Good Morning-
I am looking for some type of gel pad to place under the microtome (or perhaps under the waterbath...) to help with vibration - the water in the waterbath is jumping all over the place. Both instruments are new and the waterbath is much lighter in weight than the old ones.
Thank you for your help with this!
Nancy
Dubuque, IA
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Message: 6
Date: Wed, 28 Dec 2011 08:34:07 -0600
From: "Sharon Allen" <SAllen <@t> dsmanitoba.ca>
Subject: [Histonet] FW: Quality Modules
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<BB6ADCD4B7ABB045A09A7634AC15CC610BEC824C <@t> hscxmsmx0010.ad.wrha.mb.ca>
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I am looking for information on Quality Modules that can be added into
an existing IT system and system with great Quality Modules built into
their IT platform.
Thanks
Sharon Allen
Senior Medical Technologist
Neuropathology Lab-MS435U
Health Sciences Centre
820 Sherbrook Street
Winnipeg,MB, CA
R3A 1R9
e-mail: sallen <@t> dsmanitoba.ca
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------------------------------
Message: 7
Date: Wed, 28 Dec 2011 08:40:11 -0600
From: "Sharon Allen" <SAllen <@t> dsmanitoba.ca>
Subject: [Histonet] FW: standard for sm bx microtomy
To: <histonet <@t> lists.utsouthwestern.edu>
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<BB6ADCD4B7ABB045A09A7634AC15CC610BEC824D <@t> hscxmsmx0010.ad.wrha.mb.ca>
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Hi,
Is there is a standard used for small biopsy Microtomy? I am wondering
how other sites cut their biopsies (ie the number of sections per slide
and the amount of roughing in between sections)? I am also curious
about the number of slides per biopsy.
Thanks for your help,
Sharon Allen
Senior Medical Technologist
Neuropathology Lab-MS435U
Health Sciences Centre
820 Sherbrook Street
Winnipeg,MB, CA
R3A 1R9
e-mail: sallen <@t> dsmanitoba.ca
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