Methods and Techniques Discussion => General Histology => Topic started by: SRajpara on March 06, 2015, 03:18:30 PM

Title: Trying to optimize H&E and Oil Red O stains
Post by: SRajpara on March 06, 2015, 03:18:30 PM
Hello all,

I'm trying optimize these protocols with frozen tissue sections, some containing adipose tissue. Right now, I'm having trouble keeping the tissue on the slides.

For H&E, they seem to come off after the acid/ethanol step (differentiation) and sometimes during the blueing step in the base that I'm using. My question is, should I fix the slides in formalin before staining? We keep the slides in the -20C freezer prior to staining, and lately I've been letting them air dry for 30min before beginning the stain. I use lithium carbonate for the base, and HCl in 70% ethanol for the acid/ethanol.

For ORO, tissue falls off after rinsing it in tap water after the Hematoxylin counterstain step. I use propylene glycol in the first step, and the ORO is made with 60% triethyl phosphate.

Any advice is much appreciated. Thank you!

SR
Title: Re: Trying to optimize H&E and Oil Red O stains
Post by: gula on March 07, 2015, 03:23:26 AM
The first hint is to use adhesive slides.
gula
Title: Re: Trying to optimize H&E and Oil Red O stains
Post by: SRajpara on March 08, 2015, 03:01:29 PM
I use super frost plus slides, but I'm not sure what else I should do to the slides before I section.