Hi,
I am wondering if hoescht or dapi can stain rDNA in nucleolous. Normally they just gives a hole in the nucleus indicating nucleolus.
Or if any one has a good way to visualise nucleolus activities.
Thanks
Hi Darren,
Both Hoechst and DAPI stain also rDNA. The transcriptional activity of ribosomal genes is usually very high. Thus the ratio of rDNA to (ribosomal) proteins and RNA is rather low, hence the "hole". The size of all nucleoli combined may be a good indicator for its activity. Ki-67 is a common marker to outline nucleoli. You may also try RNA polymerase I staining to label specific rDNA transcriptional activity.
Best, Lothar
Hi Darren,
This doesn't directly address your application, but for flow cytometry we use Hoechst in combination with Pyronin Y (RNA binding dye) to look at DNA and RNA content together. It's shown nicely here:
http://www.icms.qmul.ac.uk/flowcytometry/uses/cellcycleanalysis/senescence/
Lothar Schermelleh , How do we know that Hoechst can enter nucleoli and stain rDNA , which are the studies to show that?
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