[Histonet] Nile blue sulphate - methyl green

From:Yak-Nam Wang

Hello All,

I want to use the Nile blue sulphate staining method for lipids in frozen
tissue. I have two questions regarding the protocol I hope you can help

1)In the protocols that I have seen (e.g. Bancroft&Stevens Theory and
Practice of histologican techniques) it calls for the use of chloroform
washed Methyl green. After searching the histonet archives I see that methyl
green should be replaced by ethyl green, or rather methyl green is no longer
available. In switching out methyl green for ethyl green (I'm going for the
sigma ethyl green- CI 42590), aside from not needing to wash with
cholorform, are there any other protocol changes that I need to be aware of?

2)The protocols asks for frozen tissue fixed with 10% Neutral buffered
Formalin or formal-calcium. Can someone tell me if there is a difference
between using the two fixatives? I would prefer to use 10% NBF.

Thank you for your help
Yak-Nam Wang

Research Associate
Applied Physics Laboratory
University of Washington
Box 355640
Seattle WA 98195

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