[Histonet] Re: H&E Staining for SPI-Chem Low Acid GMA

From:"Hobbs, Carl"

Hi Scott,
GMA resin will take up Haemalum......you will need to differentiate the Gills in acid.
After Gill's staining:
1. Rinse in gently running tap water until clear.
2. Give slide say, 5 dips, in 0.5% HCL in distilled water, wash in tapwater 5mins ( I assume that your tapwater is slightly alkali - if not, then follow with Scott's) Examine microscopically. You will thus see if more differentiation in HCL is reqd or not.
3. Once you have optimally stained nuclei, counterstain in Eosin.
4. Wash in gently running tap water until excess eosin is removed and you are happy with balance ( this is slow, relative to alcohol, with no danger of sections becoming wrinkled because of the alcohol softening it.)
5. Carefully/firmly blot sections dry. I now leave them in a 60C oven ( only cos that's all I have) for 60 mins, minimum.
6. Place into xylene. Leave for 10 mins before mounting .
A nice easy, reliable method, imho.
NB: after cutting/mounting sections, leave in 60C oven for 2 hrs to ensure that they are "baked" on.

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