[Histonet] Methyl Green Counterstain

[Rene J Buesa]

Your problem may reside in "Step 5" (COMPATIBLE permanent mounting medium). I can think of no other cause with the protocol you describe.
  René J.

Igor Deyneko <igor.deyneko <@t> gmail.com> wrote:
  Hello.
I am trying to do a Methyl Green counterstain on some cancer tissues,
however the problem that I encounter is that the Methyl reen is quickly
washed off the slides when done according to supllied protocol.
Methyl Green is from DAKO.
Protocol:

For optimal results, use the following procedure:

1. Rehydrate tissue.

2. Stain tissue sections for 5 minutes with Methyl Green.

3. Rinse with 95% Ethanol to remove excess reagent.

4. Dehydrate through graded alcohol.

95% Ethanol 30 seconds with dipping

95% Ethanol 30 seconds with dipping

100% Ethanol 30 seconds with dipping

100% Ethanol 30 seconds with dipping

Xylene or xylene substitute 2 minutes

Xylene or xylene substitute 2 minutes

5. Mount in compatible permanent mounting media



Can anyone offer an alternative processing , maybe with different times or
alcohols?

Thank you.

Igor D.
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