Re: [Histonet] GMA 1 micron section staining

From:"Gayle Callis"

Do not rinse but put dry sections directly to Gill 3 hematoxylin 10 minutes. 
Time can vary.
Rinse 3 X with distilled water
Go directly to bluing 1 min Scotts Tap water substitute
Rinse with distilled water
Air dry sections
Eosin/phloxine for 2 to 5 minutes time will vary.
Rinse very quickly through 2 changes of 95% ethanol  only and air dry 
quickly.  I liked to use compressed or forced air,  a fan works nicely.
Coverslip with permanent mounting media over dry section.

Be aware that 1 um is really thin and will not look like a regular H&E on a 
thicker section.  There just isn't much tissue in a 1 um thick section.  I 
had pathologists complain, but when this was explained, they understood what 
was going on since they were used to looking at 5 um thick sections.

Alcohols during dehyration will cause sections to release or bulge from 
slide.  Xylene can create problems too so we always mounted a coverglass 
over a dry section.   You can thin the mounting media a bit just in case it 
doesn't flow well.  If you have a bit of funky plastic fold over of plastic, 
that can be scraped away with a teflon coated razor blade, to not have extra 
thickness in a bad place during coverslipping.

We did not use Harris hematoxylin for GMA sections, and this method 
basically is what Polysciences put out years ago when GMA first came out, 
and it worked very well for us over the years.

Good luck on staining

Gayle M. Callis

----- Original Message ----- 
From: "Masterson_John" 
To: "Anthony Boris" ; 

Sent: Thursday, August 07, 2008 7:33 AM
Subject: [Histonet] GMA 1 micron section staining

Hello Histonetters,

I need to stain some 1 micron sections of GMA embedded tissue for H&E.
Does anyone have experience they can share?  Thanks in advance.


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