RE: staining of free floating sections

From:"Marinos, Nancy (NIH/NIDCR)"

Hi Melissa,
I have done this staining years ago.  I cut frozen sections and floated them
into Cell well plates filled with PBS.  I Fix them  and  did special stains
on them and also immunohistochemistry .  All staining was done in the wells.
You can use a pipette to change the reagents or hook up a pipette to a
vacuum line and use that to remove the reagents and a pipetter to replace
reagents.  When the staining is complete you very carefully remove the
section with a brush or brushes and place it on the slide.  The immuno
staining worked very well.  Hope this helps.  Nancy

> ----------
> From: 	Melissa Jans
> Sent: 	Tuesday, October 29, 2002 2:39 PM
> To:
> Subject: 	staining of free floating sections
> I have some 100, 200 and 400 micron thick lung sections that a pathologist
> would like to have stained.  These sections are loose (not adhered to a
> slide) and they do not have paraffin in them (don't ask me how they did
> this but they did).  He would like me to do an H&E, Alcian Blue/PAS,
> Trichrome, and and Elastichrome.  Does anyone have experience doing this?
> What is the best way to transfer your loose section from reagent to
> reagent?  Do I need to dilute my solutions and/or change my staining
> times?  
> Any help would be appreciated, this is something totally new to me.
> Thanks!!
> Melissa Jans
> Lead Scientist, Histology Lab
> University of Iowa Healthcare
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