Author Topic: Trying to optimize H&E and Oil Red O stains  (Read 1253 times)

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Offline SRajpara

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Trying to optimize H&E and Oil Red O stains
« 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

Trying to optimize H&E and Oil Red O stains
« on: March 06, 2015, 03:18:30 PM »

Offline gula

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Re: Trying to optimize H&E and Oil Red O stains
« Reply #1 on: March 07, 2015, 03:23:26 AM »
The first hint is to use adhesive slides.
gula

Offline SRajpara

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Re: Trying to optimize H&E and Oil Red O stains
« Reply #2 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.

Re: Trying to optimize H&E and Oil Red O stains
« Reply #2 on: March 08, 2015, 03:01:29 PM »