Re: methyl green as counterstain
From: | Robert Schoonhoven <rschoonh@sph.unc.edu> |
Tina,
The following has worked very well for me over the years.
Methyl Green Counterstaining Procedure
Reagents:
0.1 N Acetic Acid
0.1 N Sodium Acetate
Methyl Green
Buffer:
Add 755 ml of 0.1 N acetic acid to 264 ml of sodium acetate and adjust
the pH to 4.2 with NAOH
To the above add 20 gm Methyl Green.
Can be stored at room temp.
If staining turns bluish than the solution has started to deteriorate.
Staining:
1- Rinse in distilled water briefly
2- Place slides in methyl green for 3-5 minutes (or longer )
3- Do NOT rinse with water. Go directly into 95 % EtoH for about 10
dips, then continue through 1 more 95% and 3 100% EtoH changes leaving
the slides in the last absolute for 3 - 5 minutes.
4- Clear with 2-3 changes of xylene and coverslip
Tina Cardamone wrote:
>
> Hello,
>
> I have a simple question regarding methyl green.
> Does anyone have a methyl green counterstain protocol for
> immunohistochemistry.
> It sounds simple however I can't seem to find a protocol anywhere.
>
> Thanks in advance.
>
> Tina Cardamone
> University of Melbourne.
--
best regards,
Bob
Robert Schoonhoven
Laboratory of Molecular Carcinogenesis and Mutagenesis
Dept. of Environmental Sciences and Engineering
University of North Carolina
CB#7400
Chapel Hill, NC 27599
Phone
office 919-966-6343
Lab 919-966-6140
Fax 919-966-6123
Don't go around saying the world owes you a living; the world owes you
nothing; it was here first.
Mark Twain [Samuel Langhornne Clemens] (1835-1910)
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