fluorescence counter -stain
|From:||Angeline Martin-Studdard <AMARTIN@mail.mcg.edu>|
I do mostly fluorescent staining and I really like the results I get with a Hoechst counter-stain. This stains the nuclei of all normal cells a light blue which is visible through a DAPI filter. It provides nice contrast with the typical fluorescent visualization signals used.
The procedure I use is very simple.
1)Make up a stock solution of 10mg/ml bis-benzimide. (keep refrigerated for future use)
2)Use this stock to make up a "working" solution. Dilute bis-benzimide 1:12,000 in 1X PBS.
3)After you have completed your antibody staining, immerse your slides in the "working" solution for 8 min.
4)Wash slides in buffer and coverslip.
I usually keep refrigerated and reuse the "working" solution for a month. Bis-benzimide is a carcinogenic so use proper precautions.
Hope this helps,
medical college of georgia
>Date: 13 Feb 2001 09:26:08 -0600
>From: "Patrick M. Haley " <email@example.com>
>Subject: fluorescence counter-stain
I have a very novice understanding of the techniques used in this area,
and I have a researcher who would like to know that is the counter-stain of
choice for immuno-fluorescence. I have spoken to some other techs, and they
do not use a counterstain.
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