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DAPI/Hoechst staining live yeast cellsHey, I am looking for a protocol to stain

DAPI yeast cerevisiae live imaging Hoechst

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#1 Alfahelix

Alfahelix

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Posted 06 July 2014 - 02:33 AM

Hey,

I am looking for a protocol to stain live yeast cells with DAPI or Hoechst In 1x PBS or 1x PBS with 2 % glucose.

 

So far I have tried using 1 μl of DAPI [1 mg/ml] in 1 ml cell suspension and incubated for 10 min at RT. Washed 3x times prior to imaging. Weirdly enough, staining in 1x PBS with 2 % glucose gave better results than just using PBS. However, the cells were over stained with dots appearing at cell periphery. What

 

For the Hoescht staining I have problems with the whole cell being stained and not being able to get a nice nuclear staining. Here I have tried various concentrations in the range of 2.5-10.5 μg/ml but I have not figured out any pattern.

 

So, which factor(s) should I improve to get a nicer staining; concentration or incubation time?

Also, which staining percentage can one expect for Hoechst staining?  Does Hoechst staining include the nucleolus or just the nucleoplasm?

 

Thank you!







Also tagged with one or more of these keywords: DAPI, yeast, cerevisiae, live imaging, Hoechst

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