[Histonet] RE: IHC screening on TMA slides

From:"Langxing Pan"

Hi Ana,

It is extremely hard to make the TMA for IHC screening of 100 hybridomas
simultaneously on one slide.  I agree with Patsy that ELISA/IHC(up to 380
wells or samples per plate) would be the solution for real high throughput.
This technique has been used to screen hybridomas on cells. You may just
adapt it for tissue fragments.

I used to use multi-well slides for my hybridoma IHC screening. I put two
tissues per well. Each slide contains 10 wells. That was done before the TMA
technology. Erie Scientific (now sold as Thermo Scientific) still sells
these slides (http://www.eriesci.com/diagnostic/excell_slides.aspx). However
it is very hard and time-consuming to pick sections onto each well on the
slide.

We recently made a TMA for a company for antibody screening, which contained
eight 5mm cores. Each 5mm core contained five tissues of 1 mm core size. I
think that it is possible to make up to 20 cores of  1 ~ 1.5mm/per slide
that are separated well  enough for antibody screening, or 8 big cores/per
slide. Each big core can contain up to nine 1mm tissue elements. If you need
any help about similar TMA, do let me know.

All the best,


Langxing Pan, M.D., Ph.D.
Pantomics, Inc.
457 Mariposa Street
CA 94107, U.S.A.
Direct line: 1-510-207-9788
Fax: 1-510-653-1227
www.pantomics.com



Date: Fri,  2 Mar 2007 18:57:45 +0100 (CET)
From: Ana MERINO-TRIGO 
Subject: [Histonet] IHC screening on TMA slides
To: "histonet@lists.utsouthwestern.edu"
	
Message-ID: <16856336.81091172858265135.JavaMail.www@wwinf1606>
Content-Type: text/plain; charset=UTF-8

 
Hi Histonet, 
 
I was wondering if anyone could give me any ideas or suggestions in order to
perform IHC screening over a TMA slide containing about 100 spots of 0.6 or
1.0 mm of the same tissue. The idea it will be to select hybridomes for
further characterization based on IHC reactivity on a given positive control
tissue.  Around 100 hybridomas to test on a TMA  slide containing  spots
from the same sample. I cannot see the way to do the immuno without spread
the antibody in between the spots. I heard that exits multispot type of lids
that you could put over the slide to perform the immno but I don't really
know where to look for these lids or if they are good enough to be sure that
antibody doesn't spread in between spots. 
 
Any information it will be really appreciate it, 
 
All the best,
Ana


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