In the MCF7 human breast cancer cell line, I noticed that cells which highly expressed my GFP-tagged protein of interest have much dimmer nuclear staining with Hoechst dye, to the point where you almost cannot see them by eye in the images. I briefly searched the literature and couldn't find any reasons why this may be. Does anyone know what could be happening?
Which Hoechst stain are you using? Does it happen to emit at ~400? If so, the GFP will absorb the emission of the Hoechst. Try DAPI and see if that fixes the problem.
These are both interesting possibilities.
Daniel Lee Adams- I'm using the common Hoechst 33342 stain for DNA labeling in live cells. Its ex/em is 360/460nm and my tag is eGFP, which is excited at 488nm so your explanation seems likely. I will try another nuclear stain to confirm.
Thank you all!
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