[Histonet] Staining coral tissues embedded in resin

Laura Hunt lhunt <@t> uta.edu
Wed Feb 6 09:20:17 CST 2008


Hi everyone,


I am new to the forum.  I am working on coral tissues embedded in  
Immunobed resin and paraffin.  We would like to stain both with H&E  
and Azure II /Basic Fuchsin.  I have never tried staining plastic with  
H&E.  Any suggestions or comments would be great!  Since corals have a  
calcium carbonate skeleton, the tissues were decalcified in a EDTA  
solution.

Laura Hunt
UTA



On Feb 5, 2008, at 8:07 PM, histonet-request <@t> lists.utsouthwestern.edu  
wrote:

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> Today's Topics:
>
>   1. EpoR antibody (Colleen Forster)
>   2. RE: sections falling off (Debrosse-Serra, Beatrice)
>   3. Re: sections falling off (Rene J Buesa)
>   4. Re: Alcian Blue Quality Control OT (Robert Richmond)
>   5. HP staining (Martin, Gary)
>   6. Slide printers (ink) (Kim Merriam)
>   7. RE: Slide printers (ink) (Charles, Roger)
>   8. RE: Slide printers (ink) (Weems, Joyce)
>   9. Immunoperoxidase Double staining protocol (Reuel Cornelia)
>  10. Re: Slide printers (ink) (Patti Loykasek)
>  11. RE: Storage codes (Christine Tambasco)
>  12. Re: Slide printers (ink) (pkromund <@t> gundluth.org)
>  13. Re: Immunoperoxidase Double staining protocol (Rene J Buesa)
>  14. thanks for the support and good wishes (Sebree Linda A.)
>  15. (no subject) (Margaryan, Naira)
>  16. Do mice have tonsils?? (Colleen Forster)
>  17. eosinophils on frozen section (Breisch, Eric)
>  18. RE: eosinophils on frozen section (Ingles Claire)
>  19. A gentle reminder about using subject line Re: [Histonet] (no
>      subject) (Gayle Callis)
>  20. RE: eosinophils on frozen section (Weems, Joyce)
>  21. Re: Do mice have tonsils?? (Gayle Callis)
>  22. Re: eosinophils on frozen section (Gayle Callis)
>  23. RE: eosinophils on frozen section (Julia Dahl)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Tue, 05 Feb 2008 12:20:37 -0600
> From: Colleen Forster <cforster <@t> umn.edu>
> Subject: [Histonet] EpoR antibody
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <47A8A8F5.3090606 <@t> umn.edu>
> Content-Type: text/plain; charset=ISO-8859-1; format=flowed
>
> Hello histonetters,
>
> Is anyone doing the EpoR antibody on FFPE  for IHC? If so, could you
> share the vendor and a starting dilution.....thanks.
>
> Colleen Forster
> U of MN
>
>
>
> ------------------------------
>
> Message: 2
> Date: Tue, 5 Feb 2008 10:30:10 -0800
> From: "Debrosse-Serra, Beatrice" <Beatrice.Debrosse-Serra <@t> pfizer.com>
> Subject: RE: [Histonet] sections falling off
> To: "Olek Michalski" <olek.michalski <@t> nencki.gov.pl>,	"Histonet"
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> 	 
> <8404DFBED5207B4B8E5EEF4332CEEA5305BD4C99 <@t> lajamrexm01.amer.pfizer.com>
> Content-Type: text/plain;	charset="us-ascii"
>
> Try Newcomers silane coated slides. They are not cheap, but the  
> sections
> stay on very well.
>
> Beatrice DeBrosse-Serra
> Pathology Scientist
> Pfizer Global Research & Development
> CB4, 2150
> 10646 Science Center Drive
> San Diego, CA 92121
> Phone# 858-622-5986
> Fax# 858-678-8290
>
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Olek
> Michalski
> Sent: Tuesday, February 05, 2008 10:00 AM
> To: Histonet
> Subject: [Histonet] sections falling off
>
>
> Dear Histonetters,
>
> a friend of mine just faced a massive sections falling off. She is  
> doing
>
> Nissl staining in mouse brain (brains perfused with formalin,  
> postfixed
> 3
> days, and cryosectioned) on poly-L-lysined slides. She just managed to
> pass trough xylene and decreasing grades of alcohol to water and the
> sections started to detach. We were trying to reattach them but it  
> went
>
> out that sections are not flat any more. It seems like the tissue is
> rehydrated unevenly, but keeping it in water for hours didn't work. Is
> there any way to spread these sections not damaging them at the same
> time?
> She is likely to rescue this sections even if some work is needed.
>
> Best regards
> Olek Michalski
> -- 
>        Laboratory of Neurobiology
>       of Development and Evolution
>
>  Nencki Institute of Experimental Biology
>  ul. Pasteura 3, 02-093 Warszawa,  Poland
>  Tel. +48 22 5892268,  Fax +48 22 8225342
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 3
> Date: Tue, 5 Feb 2008 10:30:54 -0800 (PST)
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] sections falling off
> To: Olek Michalski <olek.michalski <@t> nencki.gov.pl>,	Histonet
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <87465.99172.qm <@t> web61214.mail.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> If they were cryosectioned she did not have to go through xylene or  
> graded alcohols to "dewax and hydrate" since there was no  
> dehydration or wax infiltration to begin with.
>
>  Cryosectioned sections just need to wash away the medium used to  
> cryosection (OCT perhaps?) and just stain with the aqueous solutions.
>
>  Later they can de dehydrated and cleared if a permanent mount is  
> desired.
>  Try to float the sections in a water bath and pick them up and  
> don't try again to use xylene or alcohols.
>  Probably she was following a procedure for paraffin embedded tissue  
> that should have been adapted to frozen sections and it was not.
>  René J.
>
> Olek Michalski <olek.michalski <@t> nencki.gov.pl> wrote:
>
> Dear Histonetters,
>
> a friend of mine just faced a massive sections falling off. She is  
> doing
> Nissl staining in mouse brain (brains perfused with formalin,  
> postfixed 3
> days, and cryosectioned) on poly-L-lysined slides. She just managed to
> pass trough xylene and decreasing grades of alcohol to water and the
> sections started to detach. We were trying to reattach them but it  
> went
> out that sections are not flat any more. It seems like the tissue is
> rehydrated unevenly, but keeping it in water for hours didn't work. Is
> there any way to spread these sections not damaging them at the same  
> time?
> She is likely to rescue this sections even if some work is needed.
>
> Best regards
> Olek Michalski
> -- 
> Laboratory of Neurobiology
> of Development and Evolution
>
> Nencki Institute of Experimental Biology
> ul. Pasteura 3, 02-093 Warszawa, Poland
> Tel. +48 22 5892268, Fax +48 22 8225342
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ---------------------------------
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> Search.
>
> ------------------------------
>
> Message: 4
> Date: Tue, 5 Feb 2008 13:45:34 -0500
> From: "Robert Richmond" <RSRICHMOND <@t> aol.com>
> Subject: [Histonet] Re: Alcian Blue Quality Control OT
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> 	<abea52a60802051045r44b6abeeua2f68125ea25c097 <@t> mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> Claire Ingles remarks: So that's what GOP means!
>
> Not this grumpy old pathologist - I voted for Barack Obama!
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
>
>
>
> ------------------------------
>
> Message: 5
> Date: Tue, 5 Feb 2008 11:00:57 -0800
> From: "Martin, Gary" <gmartin <@t> marshallmedical.org>
> Subject: [Histonet] HP staining
> To: <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> 	<6ED9D4252F278841A0593D3D788AF24C01ACDF72 <@t> mailsvr.MARSHMED.local>
> Content-Type: text/plain;	charset="us-ascii"
>
> To all who responded to my e-mail concerning rapid HP staining to
> include a yellow back ground ... thank you very much ... my question  
> was
> answered.
>
> Gray
>
>
>
> ------------------------------
>
> Message: 6
> Date: Tue, 5 Feb 2008 11:48:08 -0800 (PST)
> From: Kim Merriam <kmerriam2003 <@t> yahoo.com>
> Subject: [Histonet] Slide printers (ink)
> To: Histonet <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <813260.30617.qm <@t> web50306.mail.re2.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Hi everyone,
>
>  I was wondering what everyone's experience was with the TBS slide  
> printer (Shurmark-plus), the one that prints with ink?  I would  
> appreciate input, good or bad.
>
>  We currently have an etcher, and we hate it, so we are looking to  
> trade it in for something that uses ink.  I have used the Leica  
> printer in the past and have had reasonably good luck with it, but I  
> was wondering about the printer from TBS, I don't know anyone that  
> has one.
>
>  Kim
>
>
> Kim Merriam, MA, HT(ASCP)
> Cambridge, MA
>
> ---------------------------------
> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.   
> Try it now.
>
> ------------------------------
>
> Message: 7
> Date: Tue, 5 Feb 2008 14:58:42 -0500
> From: "Charles, Roger" <rcharles <@t> state.pa.us>
> Subject: RE: [Histonet] Slide printers (ink)
> To: "Kim Merriam" <kmerriam2003 <@t> yahoo.com>
> Cc: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> 	<12E4E17FEF6EBE4BAE95BEB3CDCB57000943BB6E <@t> enhbgpri04.backup>
> Content-Type: text/plain;	charset="US-ASCII"
>
> Hello,
> Recently we bought a sakura slide writer that uses ink to print. The
> only problem I have found with the ink is if you put labels over the
> print and then try to remove the label to verify a number error, or
> something like that, the ink will actually come off with the label.   
> If
> you can read upside down you can still verify the number.  I did run
> this ink thru a battery of chemicals including straight formic acid  
> for
> 5 minutes and the ink did not leave the slide.  I would caution in
> buying a Sakura slide write as it has a very unfriendly Access based
> program and the writer does not work with charged slides.  This has  
> been
> verified by me and an independent engineer sent to look at this issue.
> I do use a TBS cassette writer and love that so if I can get my sakura
> sent back I will be looking at TBS's slide writer too.
> Good luck
> Roger
>
> Roger Charles
> Microbiologist
> Pennsylvania Veterinary Laboratory
> 2305 N Cameron St
> Harrisburg, PA 17110
> 717-787-8808
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim
> Merriam
> Sent: Tuesday, February 05, 2008 2:48 PM
> To: Histonet
> Subject: [Histonet] Slide printers (ink)
>
> Hi everyone,
>
>  I was wondering what everyone's experience was with the TBS slide
> printer (Shurmark-plus), the one that prints with ink?  I would
> appreciate input, good or bad.
>
>  We currently have an etcher, and we hate it, so we are looking to
> trade it in for something that uses ink.  I have used the Leica  
> printer
> in the past and have had reasonably good luck with it, but I was
> wondering about the printer from TBS, I don't know anyone that has  
> one.
>
>  Kim
>
>
> Kim Merriam, MA, HT(ASCP)
> Cambridge, MA
>
> ---------------------------------
> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.   
> Try
> it now.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 8
> Date: Tue, 5 Feb 2008 15:25:58 -0500
> From: "Weems, Joyce" <JWEEMS <@t> sjha.org>
> Subject: RE: [Histonet] Slide printers (ink)
> To: "Charles, Roger" <rcharles <@t> state.pa.us>,	"Kim Merriam"
> 	<kmerriam2003 <@t> yahoo.com>
> Cc: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> 	<1CD6831EB9B26D45B0A3EAA79F7EBD320518E54D <@t> sjhaexc02.sjha.org>
> Content-Type: text/plain;	charset="us-ascii"
>
> We have Sakura cassette and slide writers. We print charged slides
> daily.
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of  
> Charles,
> Roger
> Sent: Tuesday, February 05, 2008 2:59 PM
> To: Kim Merriam
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] Slide printers (ink)
>
> Hello,
> Recently we bought a sakura slide writer that uses ink to print. The
> only problem I have found with the ink is if you put labels over the
> print and then try to remove the label to verify a number error, or
> something like that, the ink will actually come off with the label.   
> If
> you can read upside down you can still verify the number.  I did run
> this ink thru a battery of chemicals including straight formic acid  
> for
> 5 minutes and the ink did not leave the slide.  I would caution in
> buying a Sakura slide write as it has a very unfriendly Access based
> program and the writer does not work with charged slides.  This has  
> been
> verified by me and an independent engineer sent to look at this issue.
> I do use a TBS cassette writer and love that so if I can get my sakura
> sent back I will be looking at TBS's slide writer too.
> Good luck
> Roger
>
> Roger Charles
> Microbiologist
> Pennsylvania Veterinary Laboratory
> 2305 N Cameron St
> Harrisburg, PA 17110
> 717-787-8808
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim
> Merriam
> Sent: Tuesday, February 05, 2008 2:48 PM
> To: Histonet
> Subject: [Histonet] Slide printers (ink)
>
> Hi everyone,
>
>  I was wondering what everyone's experience was with the TBS slide
> printer (Shurmark-plus), the one that prints with ink?  I would
> appreciate input, good or bad.
>
>  We currently have an etcher, and we hate it, so we are looking to
> trade it in for something that uses ink.  I have used the Leica  
> printer
> in the past and have had reasonably good luck with it, but I was
> wondering about the printer from TBS, I don't know anyone that has  
> one.
>
>  Kim
>
>
> Kim Merriam, MA, HT(ASCP)
> Cambridge, MA
>
> ---------------------------------
> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.   
> Try
> it now.
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
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> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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> ------------------------------
>
> Message: 9
> Date: Tue, 05 Feb 2008 14:34:27 -0600
> From: "Reuel Cornelia" <Reuel.Cornelia <@t> tsrh.org>
> Subject: [Histonet] Immunoperoxidase Double staining protocol
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <47A873F3020000C50002ADF9 <@t> nwcl02.tsrh.org>
> Content-Type: text/plain; charset=US-ASCII
>
> I was asked to do an immunoperoxidase double staining on CD61 ,Cd42,  
> pf4 with an unknown mouse antibody that will work on platelets. Can  
> somebody help me or any protocol available that I can refer to. I am  
> used of IF double stianing but not Immunoperoxidase. Thank you.
>
> Reuel Cornelia, BS MT, AMT
> Cellular Pathology
> Texas Scottish Rite Hospital for Children
> 2222 Welborn Street
> Dallas, TX 75219
> Tel: 214-559-7766
> fax: 214-559-7768
>
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> ------------------------------
>
> Message: 10
> Date: Tue, 05 Feb 2008 12:39:50 -0800
> From: Patti Loykasek <ploykasek <@t> phenopath.com>
> Subject: Re: [Histonet] Slide printers (ink)
> To: "Charles, Roger" <rcharles <@t> state.pa.us>,	Kim Merriam
> 	<kmerriam2003 <@t> yahoo.com>
> Cc: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <C3CE0996.12D0C%ploykasek <@t> phenopath.com>
> Content-Type: text/plain;	charset="US-ASCII"
>
> Roger- We have been using a sakura slide printer for a couple of  
> years. I do
> agree with your comments about the software - not user friendly at  
> all nor
> is it intuitive. I have used Access database in the past without these
> issues. I do not know about the labels, I don't think we've had that  
> issue.
> Maybe it varies with the label. I don't quite understand your  
> comment on
> charged slides, as all of the slides we put thru the printer are  
> charged and
> we have no issues with that. We use charged slides (Okando Plus)  
> from BBC
> and Probe-On Plus slides from ThermoFisher. What brand of slides  
> were you
> using & what issue did you have with the slide printer from the usage?
>
>
> Patti Loykasek BS, HTL, QIHC
> PhenoPath Laboratories
> Seattle, WA
>
>
>
>
>
>
>
>> Hello,
>> Recently we bought a sakura slide writer that uses ink to print. The
>> only problem I have found with the ink is if you put labels over the
>> print and then try to remove the label to verify a number error, or
>> something like that, the ink will actually come off with the  
>> label.  If
>> you can read upside down you can still verify the number.  I did run
>> this ink thru a battery of chemicals including straight formic acid  
>> for
>> 5 minutes and the ink did not leave the slide.  I would caution in
>> buying a Sakura slide write as it has a very unfriendly Access based
>> program and the writer does not work with charged slides.  This has  
>> been
>> verified by me and an independent engineer sent to look at this  
>> issue.
>> I do use a TBS cassette writer and love that so if I can get my  
>> sakura
>> sent back I will be looking at TBS's slide writer too.
>> Good luck
>> Roger
>>
>> Roger Charles
>> Microbiologist
>> Pennsylvania Veterinary Laboratory
>> 2305 N Cameron St
>> Harrisburg, PA 17110
>> 717-787-8808
>>
>> -----Original Message-----
>> From: histonet-bounces <@t> lists.utsouthwestern.edu
>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim
>> Merriam
>> Sent: Tuesday, February 05, 2008 2:48 PM
>> To: Histonet
>> Subject: [Histonet] Slide printers (ink)
>>
>> Hi everyone,
>>
>> I was wondering what everyone's experience was with the TBS slide
>> printer (Shurmark-plus), the one that prints with ink?  I would
>> appreciate input, good or bad.
>>
>> We currently have an etcher, and we hate it, so we are looking to
>> trade it in for something that uses ink.  I have used the Leica  
>> printer
>> in the past and have had reasonably good luck with it, but I was
>> wondering about the printer from TBS, I don't know anyone that has  
>> one.
>>
>> Kim
>>
>>
>> Kim Merriam, MA, HT(ASCP)
>> Cambridge, MA
>>
>> ---------------------------------
>> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.   
>> Try
>> it now.
>> _______________________________________________
>> 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
>>
>
>
>
> This e-mail message, including any attachments, is for the sole use  
> of the
> intended recipients and may contain privileged information. Any  
> unauthorized
> review, use, disclosure or distribution is prohibited. If you are  
> not the intended
> recipient, please contact the sender by e-mail and destroy all  
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> at (206) 374-9000.
>
>
>
>
> ------------------------------
>
> Message: 11
> Date: Tue, 5 Feb 2008 21:10:37 +0000
> From: Christine Tambasco <immrstambo <@t> hotmail.com>
> Subject: RE: [Histonet] Storage codes
> To: Barbara Albert <barbaraaalbert <@t> yahoo.com>, histonet
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <BLU120-W20774ADFA25B60B43F92CAD22C0 <@t> phx.gbl>
> Content-Type: text/plain; charset="iso-8859-1"
>
>
> I always kept my gold chloride in the refrigerator. Yours is in a  
> vial though so is it a powder?
> That is what I used to have I kept at room temperature.
> Just my advice.
> Christine Tambasco, HT (ASCP)
> St. Marys Hospital
> Amsterdam, NY> Date: Mon, 4 Feb 2008 15:33:30 -0800> From: barbaraaalbert <@t> yahoo.com 
> > To: histonet <@t> lists.utsouthwestern.edu> Subject: [Histonet] Storage  
> codes> > Hi all,> I just received some vials of Gold Chloride that I  
> had ordered and was checking the label for stoarge requirements. It  
> has "Storage Code White". I haven't heard of storage codes and want  
> to know where I can go to find out what they mean.> > Thanks,>  
> Barbara Albert> UCSF Medical Center> San Francisco> > >  
> ---------------------------------> Never miss a thing. Make Yahoo  
> your homepage.> _______________________________________________>  
> Histonet mailing list> Histonet <@t> lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> _________________________________________________________________
> Climb to the top of the charts! Play the word scramble challenge  
> with star power.
> http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_jan
>
> ------------------------------
>
> Message: 12
> Date: Tue, 5 Feb 2008 15:34:42 -0600
> From: pkromund <@t> gundluth.org
> Subject: Re: [Histonet] Slide printers (ink)
> To: Patti Loykasek <ploykasek <@t> phenopath.com>
> Cc: histonet <@t> lists.utsouthwestern.edu,
> 	histonet-bounces <@t> lists.utsouthwestern.edu, "Charles,	Roger"
> 	<rcharles <@t> state.pa.us>
> Message-ID:
> 	<OF3DEC5571.973DF9B7-ON862573E6.00759A05-862573E6.007688A6 <@t> gundluth.org 
> >
> 	
> Content-Type: text/plain; charset=US-ASCII
>
> Do any of these slide printers affect precut control tissue on the  
> charged
> slides?
> Pamela
>
>
>
>             Patti Loykasek
>             <ploykasek <@t> phenop
>              
> ath.com>                                                   To
>             Sent by:                  "Charles, Roger"
>             histonet-bounces@         <rcharles <@t> state.pa.us>, Kim  
> Merriam
>             lists.utsouthwest         <kmerriam2003 <@t> yahoo.com>
>              
> ern.edu                                                    cc
>                                        
> histonet <@t> lists.utsouthwestern.edu
>                                                                    
> Subject
>             02/05/2008 02:39          Re: [Histonet] Slide printers  
> (ink)
>             PM
>
>
>
>
>
>
>
>
>
> Roger- We have been using a sakura slide printer for a couple of  
> years. I
> do
> agree with your comments about the software - not user friendly at  
> all nor
> is it intuitive. I have used Access database in the past without these
> issues. I do not know about the labels, I don't think we've had that  
> issue.
> Maybe it varies with the label. I don't quite understand your  
> comment on
> charged slides, as all of the slides we put thru the printer are  
> charged
> and
> we have no issues with that. We use charged slides (Okando Plus)  
> from BBC
> and Probe-On Plus slides from ThermoFisher. What brand of slides  
> were you
> using & what issue did you have with the slide printer from the usage?
>
>
> Patti Loykasek BS, HTL, QIHC
> PhenoPath Laboratories
> Seattle, WA
>
>
>
>
>
>
>
>> Hello,
>> Recently we bought a sakura slide writer that uses ink to print. The
>> only problem I have found with the ink is if you put labels over the
>> print and then try to remove the label to verify a number error, or
>> something like that, the ink will actually come off with the  
>> label.  If
>> you can read upside down you can still verify the number.  I did run
>> this ink thru a battery of chemicals including straight formic acid  
>> for
>> 5 minutes and the ink did not leave the slide.  I would caution in
>> buying a Sakura slide write as it has a very unfriendly Access based
>> program and the writer does not work with charged slides.  This has  
>> been
>> verified by me and an independent engineer sent to look at this  
>> issue.
>> I do use a TBS cassette writer and love that so if I can get my  
>> sakura
>> sent back I will be looking at TBS's slide writer too.
>> Good luck
>> Roger
>>
>> Roger Charles
>> Microbiologist
>> Pennsylvania Veterinary Laboratory
>> 2305 N Cameron St
>> Harrisburg, PA 17110
>> 717-787-8808
>>
>> -----Original Message-----
>> From: histonet-bounces <@t> lists.utsouthwestern.edu
>> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim
>> Merriam
>> Sent: Tuesday, February 05, 2008 2:48 PM
>> To: Histonet
>> Subject: [Histonet] Slide printers (ink)
>>
>> Hi everyone,
>>
>> I was wondering what everyone's experience was with the TBS slide
>> printer (Shurmark-plus), the one that prints with ink?  I would
>> appreciate input, good or bad.
>>
>> We currently have an etcher, and we hate it, so we are looking to
>> trade it in for something that uses ink.  I have used the Leica  
>> printer
>> in the past and have had reasonably good luck with it, but I was
>> wondering about the printer from TBS, I don't know anyone that has  
>> one.
>>
>> Kim
>>
>>
>> Kim Merriam, MA, HT(ASCP)
>> Cambridge, MA
>>
>> ---------------------------------
>> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.   
>> Try
>> it now.
>> _______________________________________________
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>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>> _______________________________________________
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>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>
>
>
> This e-mail message, including any attachments, is for the sole use  
> of the
> intended recipients and may contain privileged information. Any
> unauthorized
> review, use, disclosure or distribution is prohibited. If you are  
> not the
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>
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>
> ------------------------------
>
> Message: 13
> Date: Tue, 5 Feb 2008 13:46:41 -0800 (PST)
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] Immunoperoxidase Double staining protocol
> To: Reuel Cornelia <Reuel.Cornelia <@t> tsrh.org>,
> 	histonet <@t> lists.utsouthwestern.edu
> Message-ID: <784337.61146.qm <@t> web61225.mail.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Generally speaking you will have to run the whole protocol for one  
> of the Abs and use one chromogen (of your selection). After that,  
> since you already did HIER and blocked the internal peroxidase, you  
> will treat the section with the second Ab, will link/detect it and  
> use another chromogen that ideally should have a different color.
>  The first chromogen could be regular DAB and for the second you can  
> use also DAB but with incorporated nickel that will give a deep/blue  
> purple color, in contrast with the brown/reddish normal color of the  
> DAB.
>  The thing is that you don't need any special protocol, just your  
> regular IHC protocol for FFPE tissue, run twice with two different  
> chromogens.
>  René J.
>
> Reuel Cornelia <Reuel.Cornelia <@t> tsrh.org> wrote:
>  I was asked to do an immunoperoxidase double staining on  
> CD61 ,Cd42, pf4 with an unknown mouse antibody that will work on  
> platelets. Can somebody help me or any protocol available that I can  
> refer to. I am used of IF double stianing but not Immunoperoxidase.  
> Thank you.
>
> Reuel Cornelia, BS MT, AMT
> Cellular Pathology
> Texas Scottish Rite Hospital for Children
> 2222 Welborn Street
> Dallas, TX 75219
> Tel: 214-559-7766
> fax: 214-559-7768
>
> *******************************************************************************************************************
> Texas Scottish Rite Hospital for Children is one of the nation's  
> leading pediatric centers for the
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> Try it now.
>
> ------------------------------
>
> Message: 14
> Date: Tue, 5 Feb 2008 15:47:03 -0600
> From: "Sebree Linda A." <LSebree <@t> uwhealth.org>
> Subject: [Histonet] thanks for the support and good wishes
> To: "Histonet" <histonet <@t> pathology.swmed.edu>
> Message-ID:
> 	<D7336540A20FB0479306D82CE05B8086011F5D9B <@t> uwhis-xchng4.uwhis.hosp.wisc.edu 
> >
> 	
> Content-Type: text/plain;	charset="US-ASCII"
>
> Hi all,
>
> Well apparently our CAP inspector took the word of our supervisor on
> everything pertaining to our lab, reviewed some of our slides and was
> out the door before the weather associated with the "winter storm
> warning" arrived!  Some times its good to live in the land of ice and
> snow!
>
> Thanks for all the good luck wishes!
>
> Linda Sebree, HT(ASCP)
> University of Wisconsin Hospital & Clinics
> IHC/ISH Laboratory
> A4/204-3224
> 600 Highland Ave.
> Madison, WI 53792
> (608)265-6596
> FAX: (608)262-7174
>
>
>
> ------------------------------
>
> Message: 15
> Date: Tue, 5 Feb 2008 16:50:57 -0600
> From: "Margaryan, Naira" <NMargaryan <@t> childrensmemorial.org>
> Subject: [Histonet] (no subject)
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> 	<EE5AF506236CEE4CAFD1A281F5DCFE90014128F1 <@t> CMHEXC02EVS.childrensmemorial.org 
> >
> 	
> Content-Type: text/plain;	charset="us-ascii"
>
> Dear histo team,
>
>
>
> I need to stain frozen & FFPE sections for Tuj-1 (AbCam, ab53234) and
> FFPE sectiond for Netrin-1 (AbCam, ab39370). Any kind of suggestions  
> or
> even full IHC or IF protocol is appreciated.
>
>
>
> Thanks in advance,
>
> Naira
>
>
>
>
>
>
>
>
>
> ------------------------------
>
> Message: 16
> Date: Tue, 05 Feb 2008 17:36:48 -0600
> From: Colleen Forster <cforster <@t> umn.edu>
> Subject: [Histonet] Do mice have tonsils??
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <47A8F310.20905 <@t> umn.edu>
> Content-Type: text/plain; charset=ISO-8859-1; format=flowed
>
> For any of you have done mouse necropsy, do they have tonsils or just
> lymph nodes? I have had a request for mouse tonsils.... can't seem to
> find them in my searches. I would appreciate any help on this one.
>
> Thanks,
>
> Colleen Forster
> U of MN
>
>
>
> ------------------------------
>
> Message: 17
> Date: Tue, 5 Feb 2008 15:32:31 -0800
> From: "Breisch, Eric" <ebreisch <@t> rchsd.org>
> Subject: [Histonet] eosinophils on frozen section
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> 	<43B97B4C402C2C44AAA2A8D2C86A88B31A5A78 <@t> e2k3backend1.RCHSD.org>
> Content-Type: text/plain;	charset="US-ASCII"
>
> To all interested Histonetters:
>
>
>
>
>
> We are experiencing some frustration with our ability to identify
> eosinophils from frozen sections. For some reason which our frozen
> section set up does not enable us to identify eosinophils. Eosinophil
> identification is not impaired when the tissue is submitted for
> permanents. Could some one please share the reagents used and how the
> frozen section staining area is set up if they have success  
> identifying
> eosinophils from a frozen section? We have had many frozen sections  
> done
> on fresh tissues suspicious for an eosinophilic granuloma but our
> techniques so far have not rendered consistent eosinophil
> identification. We presently are using 95% ETOH for fixing the FS  
> slide,
> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,  
> three
> rinses in Xylene substitute and then coverslip. All other tissues  
> appear
> just fine but the eosinophils just don't show up. Any suggestions are
> greatly appreciated.
>
>
>
>
>
> Eric A. Breisch, Ph.D.
>
> Clinical Anatomist
>
> Dept. of Pathology
>
> Rady Children's Hospital and Health Center
>
> Associate Clinical Professor of Anatomy
>
> Dept. of Surgery
>
> UCSD School of Medicine
>
>
>
>
>
> ------------------------------
>
> Message: 18
> Date: Tue, 5 Feb 2008 18:08:15 -0600
> From: "Ingles Claire" <CIngles <@t> uwhealth.org>
> Subject: RE: [Histonet] eosinophils on frozen section
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> 	<08A0A863637F1349BBFD83A96B27A50A1200DF <@t> uwhis-xchng3.uwhis.hosp.wisc.edu 
> >
> 	
> Content-Type: text/plain;	charset="iso-8859-1"
>
> Eric:
> My guess is that the alcohol is lysing(blowing up) the unfixed  
> eosinophils, therefore they are not there to stain. Fixation  
> stabilizes the cells in order to be stained in regular paraffin  
> sections. Unfortunately, I have done a few trials using different  
> fixatives for fresh tissue to see if I could preserve these little  
> suckers. Nothing really worked satisfactorally. You might try fixing  
> in formalin for a minute or two before you continue on with your  
> stain and see if this helps. (don't forget to wash the sections  
> afterward) Hope this helps.
> Claire
>
>
>
> We are experiencing some frustration with our ability to identify
> eosinophils from frozen sections. For some reason which our frozen
> section set up does not enable us to identify eosinophils. Eosinophil
> identification is not impaired when the tissue is submitted for
> permanents. Could some one please share the reagents used and how the
> frozen section staining area is set up if they have success  
> identifying
> eosinophils from a frozen section? We have had many frozen sections  
> done
> on fresh tissues suspicious for an eosinophilic granuloma but our
> techniques so far have not rendered consistent eosinophil
> identification. We presently are using 95% ETOH for fixing the FS  
> slide,
> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,  
> three
> rinses in Xylene substitute and then coverslip. All other tissues  
> appear
> just fine but the eosinophils just don't show up. Any suggestions are
> greatly appreciated.
>
>
>
>
>
> Eric A. Breisch, Ph.D.
>
> Clinical Anatomist
>
> Dept. of Pathology
>
> Rady Children's Hospital and Health Center
>
> Associate Clinical Professor of Anatomy
>
> Dept. of Surgery
>
> UCSD School of Medicine
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 19
> Date: Tue, 5 Feb 2008 17:30:17 -0700
> From: "Gayle Callis" <gayle.callis <@t> bresnan.net>
> Subject: A gentle reminder about using subject line Re: [Histonet] (no
> 	subject)
> To: "Margaryan, Naira" <NMargaryan <@t> childrensmemorial.org>,
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000601c86857$72c69700$6601a8c0 <@t> Sunney>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> 	reply-type=original
>
> For those new to or not used to the Histonet messaging, please use the
> subject line rather ant just hit the reply key.  Replying to the daily
> digest also kicks back all the messages for that day to those who have
> already received them. The subject is important so others, including  
> me,
> don't just hit the delete key and not read your valuable inquiries and
> comment.
>
> Thanks
> Gayle M. Callis
> HTL/HT/MT(ASCP)
> Bozeman MT 59715
>
>
> ----- Original Message -----
> From: "Margaryan, Naira" <NMargaryan <@t> childrensmemorial.org>
> To: <histonet <@t> lists.utsouthwestern.edu>
> Sent: Tuesday, February 05, 2008 3:50 PM
> Subject: [Histonet] (no subject)
>
>
> Dear histo team,
>
>
>
> I need to stain frozen & FFPE sections for Tuj-1 (AbCam, ab53234) and
> FFPE sectiond for Netrin-1 (AbCam, ab39370). Any kind of suggestions  
> or
> even full IHC or IF protocol is appreciated.
>
>
>
> Thanks in advance,
>
> Naira
>
>
>
>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 20
> Date: Tue, 5 Feb 2008 19:35:12 -0500
> From: "Weems, Joyce" <JWEEMS <@t> sjha.org>
> Subject: RE: [Histonet] eosinophils on frozen section
> To: "Ingles Claire" <CIngles <@t> uwhealth.org>,
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> 	<1CD6831EB9B26D45B0A3EAA79F7EBD320518E56A <@t> sjhaexc02.sjha.org>
> Content-Type: text/plain;	charset="us-ascii"
>
> Fix in formal-alcohol (no, it is not wearing a tuxedo) 90 ml absolute
> alcohol - 10 ml 37% formaldehyde... Should work ok then... j
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ingles
> Claire
> Sent: Tuesday, February 05, 2008 7:08 PM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: RE: [Histonet] eosinophils on frozen section
>
> Eric:
> My guess is that the alcohol is lysing(blowing up) the unfixed
> eosinophils, therefore they are not there to stain. Fixation  
> stabilizes
> the cells in order to be stained in regular paraffin sections.
> Unfortunately, I have done a few trials using different fixatives for
> fresh tissue to see if I could preserve these little suckers. Nothing
> really worked satisfactorally. You might try fixing in formalin for a
> minute or two before you continue on with your stain and see if this
> helps. (don't forget to wash the sections afterward) Hope this helps.
> Claire
>
>
>
> We are experiencing some frustration with our ability to identify
> eosinophils from frozen sections. For some reason which our frozen
> section set up does not enable us to identify eosinophils. Eosinophil
> identification is not impaired when the tissue is submitted for
> permanents. Could some one please share the reagents used and how the
> frozen section staining area is set up if they have success  
> identifying
> eosinophils from a frozen section? We have had many frozen sections  
> done
> on fresh tissues suspicious for an eosinophilic granuloma but our
> techniques so far have not rendered consistent eosinophil
> identification. We presently are using 95% ETOH for fixing the FS  
> slide,
> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,  
> three
> rinses in Xylene substitute and then coverslip. All other tissues  
> appear
> just fine but the eosinophils just don't show up. Any suggestions are
> greatly appreciated.
>
>
>
>
>
> Eric A. Breisch, Ph.D.
>
> Clinical Anatomist
>
> Dept. of Pathology
>
> Rady Children's Hospital and Health Center
>
> Associate Clinical Professor of Anatomy
>
> Dept. of Surgery
>
> UCSD School of Medicine
>
>
>
> _______________________________________________
> 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
>
>
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>
> ------------------------------
>
> Message: 21
> Date: Tue, 5 Feb 2008 17:49:57 -0700
> From: "Gayle Callis" <gayle.callis <@t> bresnan.net>
> Subject: Re: [Histonet] Do mice have tonsils??
> To: "Colleen Forster" <cforster <@t> umn.edu>,
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <000e01c8685a$321bd7d0$6601a8c0 <@t> Sunney>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> 	reply-type=response
>
> Colleen,
>
> Rodents do NOT have tonsils, but have tissues analogous (Sp?) to  
> tonsils,
> called nasal associated lymphoid tissues located in a rather  
> difficult spot
> at the back of the nasal turbinates.  I suggest you get into the  
> literature
> and type in murine NALT as there are some excellent articles on  
> morphology
> and their location.  If you want to do frozen sections, you either  
> need to
> remove these from this area, not easy to do, and takes a lot of  
> practice OR
> you can do undecalcified bone frozen sections with the Cryojane.   
> The NALT
> is located above the soft palate, just below the 3rd  palatine ridge  
> if you
> start counting ridges on the soft palate and starting count at the  
> front
> incisors.  These tiny lymphoid tissues are nestled on the roof of  
> the mouth,
> just below the soft palate, between the back molars.  The molars are  
> the
> complicating factor for doing murine CD markers that only work on  
> frozen
> sections. I will be happy to send you a powerpoint photograph of a  
> fully
> decalcified mouse head, mid sagittal section, stained with H&E to show
> exactly where the NALT is located.  I also have a cross section of the
> murine nasal turbinates showing NALT in that orientation, and much  
> harder to
> deal with in order to find the NALT.  One project had PLP perfused  
> mouse
> head, immersion fixed longer and then EDTA decalcified,  
> cryoprotected and
> sectioned on the cryostat to do immunoglobulin staining of turbinate
> epithelial cells back and into the NALT.
>
> We work with NALT a good deal, and I can honestly say that
> dissection/removal is not easy.  However, it can be done but with very
> gentle, light touch using a pointed, tiny scalpel blade. When we do  
> frozen
> sections of removed NALT, we can obtain approx 30 - 40 sections,  
> serial, at
> 5 um and with treated animals to produce inflammed NALT, up to 50  
> and more
> serial sections.  With undecalcified bone, we get far fewer due to the
> difficulty of sectioning.
>
> G0 into PUBMED, and look for David Pascual, Keri Cscentis on their  
> NALT
> publication.  I believe they put a cartoon of NALT location in that
> publication.
>
> Good luck,
>
> Gayle M. Callis
> HTL/HT/MT(ASCP)
> Bozeman MT 59715
>
>
> ----- Original Message -----
> From: "Colleen Forster" <cforster <@t> umn.edu>
> To: <histonet <@t> lists.utsouthwestern.edu>
> Sent: Tuesday, February 05, 2008 4:36 PM
> Subject: [Histonet] Do mice have tonsils??
>
>
>> For any of you have done mouse necropsy, do they have tonsils or just
>> lymph nodes? I have had a request for mouse tonsils.... can't seem  
>> to find
>> them in my searches. I would appreciate any help on this one.
>>
>> Thanks,
>>
>> Colleen Forster
>> U of MN
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 22
> Date: Tue, 5 Feb 2008 17:58:05 -0700
> From: "Gayle Callis" <gayle.callis <@t> bresnan.net>
> Subject: Re: [Histonet] eosinophils on frozen section
> To: "Breisch, Eric" <ebreisch <@t> rchsd.org>,
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <001401c8685b$55212f90$6601a8c0 <@t> Sunney>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> 	reply-type=original
>
> We improved our eosinophil staining on frozen sections of murine  
> tissue by
> switching to NBF immersion fixation, let it sit 10 min, then do the  
> H&E but
> use eosin/phloxine (Richard Allan stain will work) but stain in
> eosin/phloxine for 2 minutes.    For some reason, the eosin was  
> never taken
> up as well after a shorter time in eosin alone,  and greatly  
> improved with
> the phloxine added.  Sections can be cut a bit thinner too, try 4 um  
> - it
> should make the granules more apparent against the background and  
> other
> cells. Also, I suggest adding absolute alcohol after the 95% for  
> better
> dehydration before clearing.
>
> You may have success using alcoholic formalin in order to speed up the
> fixation a bit.  However, when we switched to NBF, we had better  
> staining of
> eosinophils in frozen sections.  If you have the time, you can let NBF
> fixation go longer, we often let sections sit for days then stain,  
> but with
> your diagnostic procedure, you will want a bit more speed involved.
>
> Good luck
>
> Gayle M..Callis
> HTL/HT/MT(ASCP)
> Bozeman MT 59715
>
>
>
>
>
>
> ----- Original Message -----
> From: "Breisch, Eric" <ebreisch <@t> rchsd.org>
> To: <histonet <@t> lists.utsouthwestern.edu>
> Sent: Tuesday, February 05, 2008 4:32 PM
> Subject: [Histonet] eosinophils on frozen section
>
>
> To all interested Histonetters:
>
>
>
>
>
> We are experiencing some frustration with our ability to identify
> eosinophils from frozen sections. For some reason which our frozen
> section set up does not enable us to identify eosinophils. Eosinophil
> identification is not impaired when the tissue is submitted for
> permanents. Could some one please share the reagents used and how the
> frozen section staining area is set up if they have success  
> identifying
> eosinophils from a frozen section? We have had many frozen sections  
> done
> on fresh tissues suspicious for an eosinophilic granuloma but our
> techniques so far have not rendered consistent eosinophil
> identification. We presently are using 95% ETOH for fixing the FS  
> slide,
> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,  
> three
> rinses in Xylene substitute and then coverslip. All other tissues  
> appear
> just fine but the eosinophils just don't show up. Any suggestions are
> greatly appreciated.
>
>
>
>
>
> Eric A. Breisch, Ph.D.
>
> Clinical Anatomist
>
> Dept. of Pathology
>
> Rady Children's Hospital and Health Center
>
> Associate Clinical Professor of Anatomy
>
> Dept. of Surgery
>
> UCSD School of Medicine
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 23
> Date: Tue, 5 Feb 2008 20:06:55 -0600
> From: Julia Dahl <jdmd77 <@t> hotmail.com>
> Subject: RE: [Histonet] eosinophils on frozen section
> To: "Breisch, Eric" <ebreisch <@t> rchsd.org>,
> 	<histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <BAY105-W19998B9BBFE9D1F977234FC82D0 <@t> phx.gbl>
> Content-Type: text/plain; charset="iso-8859-1"
>
>
> Agree with Gayle that alcoholic formalin allows for speedier  
> fixation and preservation of eosinophil granule membranes (and  
> therefore staining, rather than degranulation).
>
>> Date: Tue, 5 Feb 2008 15:32:31 -0800
>> From: ebreisch <@t> rchsd.org
>> To: histonet <@t> lists.utsouthwestern.edu
>> Subject: [Histonet] eosinophils on frozen section
>>
>> To all interested Histonetters:
>>
>>
>>
>>
>>
>> We are experiencing some frustration with our ability to identify
>> eosinophils from frozen sections. For some reason which our frozen
>> section set up does not enable us to identify eosinophils. Eosinophil
>> identification is not impaired when the tissue is submitted for
>> permanents. Could some one please share the reagents used and how the
>> frozen section staining area is set up if they have success  
>> identifying
>> eosinophils from a frozen section? We have had many frozen sections  
>> done
>> on fresh tissues suspicious for an eosinophilic granuloma but our
>> techniques so far have not rendered consistent eosinophil
>> identification. We presently are using 95% ETOH for fixing the FS  
>> slide,
>> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
>> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in  
>> 70%
>> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,  
>> three
>> rinses in Xylene substitute and then coverslip. All other tissues  
>> appear
>> just fine but the eosinophils just don't show up. Any suggestions are
>> greatly appreciated.
>>
>>
>>
>>
>>
>> Eric A. Breisch, Ph.D.
>>
>> Clinical Anatomist
>>
>> Dept. of Pathology
>>
>> Rady Children's Hospital and Health Center
>>
>> Associate Clinical Professor of Anatomy
>>
>> Dept. of Surgery
>>
>> UCSD School of Medicine
>>
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _________________________________________________________________
> Connect and share in new ways with Windows Live.
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> ------------------------------
>
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>
> End of Histonet Digest, Vol 51, Issue 7
> ***************************************

Laura R Hunt, PhD
Post-doctoral associate
University of Texas at Arlington
ph: 817-272-1499
fax: 817-272-2855
lhunt <@t> uta.edu






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