[Histonet] IHC screening on TMA slides

[koellingr <@t> comcast.net]

Ana,
I thought about and briefly attempted to do something like this also when screening hybridoma's.  But I could never get past a philosophical type argument.  When I've screened for clones via FACS or ELISA or Western, I was always looking for a clone that had specificity for a restricted target.  It was on or off.  Hit or miss.  There was just target there.  But for tissue, besides your putative real target, there is a milieu of other tissue types and cells in tissue sections which can provide a great deal of information when screening uncharacterized supernatants.  You might see something very unexpected with a clone, against some other target that you would miss with TMA sized bits of tissue.  I have come across many surprises and useful info simply by seeing one clone on a large about of TMA's or tissue sections.  I would have missed those discoveries with .6 mm diameter tissue sections.  Could never have assessed the stickiness or dirtiness of any antibodies for tissues.  If you
 are looking at hundreds of supes, I'm assuming these have not even been sub-cloned yet unless you have some very energetic hybridoma people.  I think the idea of high throughput screening by applying 1 supe to one single .6 mm tissue section is indeed enticing, but I think it is hard to justify  the amount of information you could possibly loose or never discover.

Ray
Almost, but not quite yet, employed in Seattle, WA

-------------- Original message -------------- 
From: Ana MERINO-TRIGO <ana.merino-trigo <@t> wanadoo.fr> 

> Because we do want to select in a quick way the right hybridoma in between more 
> than 100 hybridomas using the same tissue. We thought in using the advantages of 
> TMA technology, but the other way around. Same tissue in multiple spots to test 
> different antibodies. So, we will need to find the way to separate physically 
> the spots to perform different immunos in the same slide. Does this make sense? 
> Thanks, 
> Ana 
> ----- Original Message ----- 
> From: Jackie M O'Connor 
> To: ana.merino-trigo <@t> wanadoo.fr 
> Cc: histonet <@t> lists.utsouthwestern.edu ; 
> histonet-bounces <@t> lists.utsouthwestern.edu 
> Sent: Friday, March 02, 2007 7:18 PM 
> Subject: Re: [Histonet] IHC screening on TMA slides 
> 
> 
> 
> I don't understand why you would have a TMA with 100 cores of the same sample. 
> 
> 
> 
> 
> Ana MERINO-TRIGO 
> Sent by: histonet-bounces <@t> lists.utsouthwestern.edu 
> 03/02/2007 11:57 AM Please respond to 
> ana.merino-trigo <@t> wanadoo.fr 
> 
> To"histonet <@t> lists.utsouthwestern.edu" 
> cc 
> Subject[Histonet] IHC screening on TMA slides 
> 
> 
> 
> 
> 
> 
> 
> 
> 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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