We want to stain the nucleus in live cells. I know that DRAQ5 can label DNA in live cells but I've heard that it can be toxic. I'd appreciate any experiences others might have had using this stain for live cell imaging.
Hi Philip,
would it be possible to tell me what was the dose of draq-5 that you used to stain your cells with? And what type of cells did you stain?
Many thanks.
Ashraf
Hi Domina, hi Philip
we were looking into using DRAQ5 as a nuclear stain for live cell imaging at a concentration of 1uM resulting in a very faint nuclear stain, 5uM gave results comparable to 1ug/ml Hoechst. It was still more toxic than any of the Syto dyes we were using or as Hoechst. Also: The DRAQ5 toxicity appeared independent of illumination while toxicity of Hoechst or Syto stained cells depends much more on the overall exposure to the required excitation light.
regards
Heiko
Hi Heiko,
do you think the effect would be the same on all cell-lines? the recommended dose is 1uM but yes it is faint as you said and if you incase the dose you will kill the cells and won't be valuable as Philip said.
Is there any reliable stain that we can use to image live cells?
Many thanks for all of you.
Ashraf
Dear Ashraf
I would recommend Hoechst (cheap) or Syto dyes if you need green, red or far red emission. But still go with lower concentrations than recommended were possible, especially for long term experiments
regards
Heiko
I guess, it depends on type of cells being stained. Probably the DRAQ5 causes changes in gene expression in next generations of Bacterial cells (E. coli and S. aureus), however, the growth and multiplication doesn't seem to be effected.
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