[Histonet] combining immunofluorescence and conventional

[Mazan, Melissa]

Hi all, 
You have to do the fluorescence last.  Almost everything else will quench the signal.  Melissa

Melissa R. Mazan, DVM, Diplomate ACVIM
Associate Professor, Director of Equine Sports Medicine
Department of Clinical Sciences
Cummings School of Veterinary Medicine
Tufts University
200 Westborough Road
North Grafton, MA, 01536
tel: 508-839-5395
email: melissa.mazan <@t> tufts.edu

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Subject: Histonet Digest, Vol 94, Issue 21

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Today's Topics:

   1. RE: Histonet Digest, Vol 94,      Issue 20 Question of combining
      immunofluorescence and more (Steve McClain)
   2. (no subject) (Teisha Robertson)
   3. Question of combining immunofluorescence and more
      conventional cytologic stains (Amos Brooks)


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Message: 1
Date: Sat, 17 Sep 2011 17:45:55 -0400
From: "Steve McClain" <SteveM <@t> mcclainlab.com>
Subject: [Histonet] RE: Histonet Digest, Vol 94,        Issue 20 Question of
        combining immunofluorescence and more
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <EE723B268CDC7F4E85D42139B6F5416337AFBB <@t> ml1.McClainLabs.local>
Content-Type: text/plain;       charset="us-ascii"

Dual Fluorescence may give you a start.
You can try Hoescht stain for nuclei and use another wavelength for the
second signal.

Eosin fluoresces , except eosin does not stain nuclei and being related
to fluoroscein eosin works on the green wavelength.

Steve
Steve A. McClain, MD
McClain Labs, LLC 45 Manor Road, Smithtown, NY 11787 631 361 4000



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Message: 2
Date: Sat, 17 Sep 2011 17:44:02 -0700 (PDT)
From: Teisha Robertson <tshrobertson <@t> yahoo.com>
Subject: [Histonet] (no subject)
To: deanl <@t> jackson.org, deanl <@t> jacksonhospital.net,
        huttowa <@t> kellyservices.com,      histonet <@t> lists.utsouthwestern.edu
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        <1316306642.97714.YahooMailMobile <@t> web126003.mail.ne1.yahoo.com>
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Message: 3
Date: Sat, 17 Sep 2011 21:13:59 -0400
From: Amos Brooks <amosbrooks <@t> gmail.com>
Subject: [Histonet] Question of combining immunofluorescence and more
        conventional cytologic stains
To: histonet <@t> lists.utsouthwestern.edu, minhan.science <@t> gmail.com
Message-ID:
        <CAC95ki826P1iniG9-AXyZs_=msQS0L1hR_E1c9fSkyVqN2tgkg <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Hi,
     Have you tried doing the conventional stain first, then the fluorescent
after. As long as the epitope survives the initial staining, it should be
fairly easy to label the cells with a fluorescent tag.

Amos

On Sat, Sep 17, 2011 at 1:00 PM,
<histonet-request <@t> lists.utsouthwestern.edu>wrote:

> Message: 5
> Date: Sat, 17 Sep 2011 11:33:29 +0800
> From: Min-Han Tan <minhan.science <@t> gmail.com>
> Subject: [Histonet] Question of combining immunofluorescence and more
>        conventional cytologic stains
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
>        <CAKdVuDQkJ2Q5icE5xxSeh6YQTbq_r+LuykxNo+hHzYBHjAcsRQ <@t> mail.gmail.com
> >
> Content-Type: text/plain; charset=ISO-8859-1
>
> Dear all,
>
> Apologies for the trouble. I am working on fresh human cancer cells on
> slides here in Singapore. I am trying to visualize the slides concurrently
> using both immunofluorescence and a more conventional nuclear stain used in
> pathology laboratories.
>
> I have tried Diffquik and Giemsa, but both seem to quench the fluorescence
> signal. I have tried a GFP-expressing cell line, and the same quenching
> seems to occur as well.
>
> I was wondering if anyone has ever needed to do similar work, and if so,
> what sort of solutions (bad pun) were adopted?
>
> Thank you!
>
> Min-Han
>


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End of Histonet Digest, Vol 94, Issue 21
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