[Histonet] xylene substitute for processing (Cross, Kelly)
Charity Wyatt
charchar999 <@t> gmail.com
Mon Mar 5 13:33:11 CST 2012
Hi, Kelly. I used Americlear at one hospital I worked at, and we had no
issues with special stains, processing, etc. We even recycled the
Americlear and re-used it with good results.
Charity Wyatt
On Mon, Mar 5, 2012 at 1:00 PM,
<histonet-request <@t> lists.utsouthwestern.edu>wrote:
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> 1. Re: Histonet Digest, Vol 100, Issue 3 (Madeleine Huey)
> 2. RE: microtomy (Maryott, Bridget)
> 3. replacing tween in the secondary hyb.solution (Geert van Haalem)
> 4. Re: replacing tween in the secondary hyb.solution (Rene J Buesa)
> 5. xylene substitute for processing (Cross, Kelly)
> 6. immunofluorescence mounting medium? (Collette, Nicole M.)
> 7. RE: immunofluorescence mounting medium? (Wen,Yujie)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Sun, 4 Mar 2012 13:18:24 -0800
> From: Madeleine Huey <madeleinehuey <@t> gmail.com>
> Subject: [Histonet] Re: Histonet Digest, Vol 100, Issue 3
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
> <CAF2e4CJMedupw409dnm70JDtHX64L9qPqKCZPv7cScAf1BrBAg <@t> mail.gmail.com
> >
> Content-Type: text/plain; charset=ISO-8859-1
>
> From: Courtney Pierce <Courtney.Pierce <@t> quintiles.com>
> Subject: [Histonet] AKT and pAKT
>
> Has anyone out there done AKT and pAKT antibodies for IHC. I have been
> working on them for about a month now and they are still not working
> right. I hope there is someone out there that can help me???
>
> Courtney,
>
> Maybe you should post your IHC protocol (AKT & pAKT) here, and
> hopefully we can find the problem & help you.
>
> I can help you as well, if you send me your protocol & IHC picture.
>
> Best!
> madeleinehuey <@t> elcaminohospital.org
>
>
>
> ------------------------------
>
> Message: 2
> Date: Mon, 5 Mar 2012 09:47:27 -0500
> From: "Maryott, Bridget" <bridget.maryott <@t> ventana.roche.com>
> Subject: [Histonet] RE: microtomy
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <
> F66E3A8D37574042826ACCBFB00C5C2102C73545A8 <@t> RNUMSEM722.nala.roche.com>
> Content-Type: text/plain; charset="us-ascii"
>
> You might want to check the tolerance of your instrument. If I remember
> correctly, even a precisely calibrated microtome can still have a range of
> around +/- 10%. So even when calibrated correctly, cutting at 3 microns and
> getting a section of 3.3 microns would still be within the manufacturer's
> specs.
>
> -Bridget Maryott, HT (ASCP)
>
> Confidentiality Note: This message is intended only for the use of the
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>
>
> Message: 1
> Date: Fri, 2 Mar 2012 12:06:43 -0600
> From: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
> Subject: [Histonet] microtomy
> To: "'histonet <@t> lists.utsouthwestern.edu'"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
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> 65365F35C0F2EF4D846EC3CA73E49C43016BE17354EC <@t> HPEMX3.HealthPartners.int>
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> Content-Type: text/plain; charset="us-ascii"
>
> We have run into an interesting scenario and wondering what the "experts"
> think! We cut bone marrow bx's and lymph nodes for lymphoma @ 3 microns on
> one particular microtome. Within the past month, the hematopathologist has
> felt the sections are thicker than the usual 3 microns. I had our service
> technician measure the microns and the equipment was cutting as set. I
> had the blocks cut on a different microtome and we have seen variations
> there also. My question is, does the amount of time on ice make a minor
> difference in the section thickness? I know a lot of responses may be the
> difference in the tech cutting inasmuch as how fast they turn the
> rotations, etc., but,we have ruled out that variable by having more than
> one tech cut at the microtome in question. I am stymied as to how to remedy
> this fluctuation! This is why we love histology, so many variables to
> create a problem and why I love histonet, so many techs to help one through
> a dilemma!! Thank you!!
>
> Dorothy Webb, HT (ASCP)
>
>
>
>
>
>
> ------------------------------
>
> Message: 3
> Date: Mon, 5 Mar 2012 15:25:17 +0000
> From: Geert van Haalem <g.vanhaalem <@t> prosensa.nl>
> Subject: [Histonet] replacing tween in the secondary hyb.solution
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <5F4E852313B6BC4B8C9916B69BAB4C1E05906E36 <@t> pro-ex02>
> Content-Type: text/plain; charset="us-ascii"
>
> Hi,
>
> I currently did an experiment in which I left out Tween in de
> hybridisation solution of
> the secondary antibodies. I only made a solution of two Alexa in PBS. When
> imaging
> Bye eye I did not notice any decrease in signal compared to a hyb.solution
> with Tween.
> My intention is to replace tween with FBS, but can anybody tell me pro and
> cons for
> this idea.
>
> With regards,
>
> Geert
>
>
> ------------------------------
>
> Message: 4
> Date: Mon, 5 Mar 2012 07:46:03 -0800 (PST)
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Subject: Re: [Histonet] replacing tween in the secondary hyb.solution
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>, Geert van Haalem
> <g.vanhaalem <@t> prosensa.nl>
> Message-ID:
> <1330962363.93868.YahooMailClassic <@t> web162105.mail.bf1.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Not a good idea. The effect of Twenn is to facilitate the dispersion over
> the section of the reagent. If you do not use it the reagents may not reach
> all the surface and you may end with an uneven reaction or your results
> could be inconsistent. A small savings by not using Tween may mean that you
> could have to repeat the test and spend much more expensive reagents.
> Again, not a good idea.
> René J.
>
> --- On Mon, 3/5/12, Geert van Haalem <g.vanhaalem <@t> prosensa.nl> wrote:
>
>
> From: Geert van Haalem <g.vanhaalem <@t> prosensa.nl>
> Subject: [Histonet] replacing tween in the secondary hyb.solution
> To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu
> >
> Date: Monday, March 5, 2012, 10:25 AM
>
>
> Hi,
>
> I currently did an experiment in which I left out Tween in de
> hybridisation solution of
> the secondary antibodies. I only made a solution of two Alexa in PBS. When
> imaging
> Bye eye I did not notice any decrease in signal compared to a hyb.solution
> with Tween.
> My intention is to replace tween with FBS, but can anybody tell me pro and
> cons for
> this idea.
>
> With regards,
>
> Geert
> _______________________________________________
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> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> ------------------------------
>
> Message: 5
> Date: Mon, 5 Mar 2012 16:24:03 +0000
> From: "Cross, Kelly" <KCross <@t> cvm.tamu.edu>
> Subject: [Histonet] xylene substitute for processing
> To: "Histonet Listserv (E-mail) (histonet <@t> lists.utsouthwestern.edu)"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
> <869848DDBB7C5D4896A569A38B814E6905576330 <@t> CVMMB01.cvm.tamu.edu>
> Content-Type: text/plain; charset="us-ascii"
>
> Greetings Histonet!
>
> Does anyone use xylene substitutes for routine over-night processing? If
> so, what do you use and does it have any adverse effect your special stains?
>
> Thank you in advance for your help,
> Kelly
>
> Kelly S. Cross B.S., HT (ASCP)
> Medical Laboratory Supervisor
> Veterinary Pathobiology
> Texas Veterinary Medical Center
> Texas A&M University
> College Station, TX 77843-4467
> 979-862-3658 Office
> 979-845-5149 Lab
>
>
>
> ------------------------------
>
> Message: 6
> Date: Mon, 5 Mar 2012 09:33:38 -0800
> From: "Collette, Nicole M." <collette2 <@t> llnl.gov>
> Subject: [Histonet] immunofluorescence mounting medium?
> To: "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <CB7A38F2.6775%collette2 <@t> llnl.gov>
> Content-Type: text/plain; charset="Windows-1252"
>
> Hello, Esteemed Histonetters,
>
> I am trying to get nice publication-quality images of my immunofluorescent
> tissue sections. I am currently using Prolong Gold, and after I let the
> stuff cure for several days, my 100X oil-immersion images are still smeary,
> even after taking into account the unevenness of the tissue section. I
> don't seem to have this problem with colorimetric stains (histological
> stains, LacZ, etc.), so I am thinking the mounting medium is part of the
> problem? It seems to me that it never fully cures at the middle of the
> coverslip… Does anyone have a recommendation for a mounting medium that
> works better for this purpose? Any advice is appreciated.
>
> Thanks in advance, and Happy Monday!
>
> Sincerely,
> Nicole Collette
> Lawrence Livermore National Laboratory
> collette2 <@t> llnl.gov
>
>
> ------------------------------
>
> Message: 7
> Date: Mon, 5 Mar 2012 17:40:26 +0000
> From: "Wen,Yujie" <yujie.wen <@t> louisville.edu>
> Subject: RE: [Histonet] immunofluorescence mounting medium?
> To: "Collette, Nicole M." <collette2 <@t> llnl.gov>,
> "histonet <@t> lists.utsouthwestern.edu"
> <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <wjwpwfroqwsqehu6dn2jl2tp.1330969083131 <@t> email.android.com>
> Content-Type: text/plain; charset="Windows-1252"
>
> http://www.vectorlabs.com/catalog.aspx?catID=279
>
> VECTASHIELD Mounting Media for Fluorescence
>
> This one works for me and was recommended by Leica technical support for
> oil immersion image.
>
> -------- Original Message --------
> From: Collette, Nicole M.
> Sent: Mon, Mar 5, 2012 12:35 PM
> To: histonet <@t> lists.utsouthwestern.edu
> CC:
> Subject: [Histonet] immunofluorescence mounting medium?
>
>
> Hello, Esteemed Histonetters,
>
> I am trying to get nice publication-quality images of my immunofluorescent
> tissue sections. I am currently using Prolong Gold, and after I let the
> stuff cure for several days, my 100X oil-immersion images are still smeary,
> even after taking into account the unevenness of the tissue section. I
> don't seem to have this problem with colorimetric stains (histological
> stains, LacZ, etc.), so I am thinking the mounting medium is part of the
> problem? It seems to me that it never fully cures at the middle of the
> coverslip… Does anyone have a recommendation for a mounting medium that
> works better for this purpose? Any advice is appreciated.
>
> Thanks in advance, and Happy Monday!
>
> Sincerely,
> Nicole Collette
> Lawrence Livermore National Laboratory
> collette2 <@t> llnl.gov
> _______________________________________________
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> Histonet <@t> lists.utsouthwestern.edu
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>
>
> ------------------------------
>
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> End of Histonet Digest, Vol 100, Issue 5
> ****************************************
>
--
Charity Wyatt
Mohs Histotechnologist
Jacksonville Skin Cancer Center, P.A./
Michael E. Lutz, M.D.
(904)737-0111
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