Immunofluorescence Nuclear Staining

[orestis]

Hiya
In tissue sections i have been trying to stain nuclei with a blue fluorophore with slides treated with 2N HCl for BrdU for an indirect immunofluorescence detection method.
Using PI, SYTOX , DAPI, and Hoeschst 33342 stains has not been successful.

Does anybody know of a good stain for denatured DNA in nuclei?

[airie]

I think one of the dyes in the Giemsa stain might by autofluorescent, and they look at Giemsa banding in chromosomes...but I do not know if it shows up in denatured DNA...

[airie]

oh haha i just realised this post was put up 3 years ago

[uwchick]

I am actually struggling with this very same issue so old post or new, its still a problem! I need a nuclear stain that will stain denatured DNA (ssDNA) so that I can also use an acid denature step to stain for BrdU using fluorescent detection. DAPI, and anything that intercalates into DNA will not work. Anyone with ideas would be greatly appreciated!

[asbess1984]

Hi uwchick,

Did you ever find a stain that works? In methods of papers with BrdU, they stain with DAPI after all of the denaturation and immunostaining when they mount the cells on a slide. Is this the way you have tried? I tried picogreen but it also is removed upon denaturation.