[Histonet] Methyl Green Counterstain

[Igor Deyneko]

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.