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Hi everybody,
I am trying to use DeltaVision environmental chamber 40X, time lapse to figure out localization of my EYFR-tagged protein. I did it once.
When I fixed cells , endogenous protein predominantly is in the nucleus and also in cytoplasm, but sometimes only in cytoplasm.
EGFP-tagged also shows the same pattern. I thought that transition could depend on stage of cells, particularly mitosis.
I am going to include DIC imaging. I also want to add DNA dye for the living cells. What would be the best choice? Or are there any other dyes to track different stages of cells?
I am using transiently transfected HPV epithelial cells which divided once in 4 days. Probably I need some synchronization before or after transfection. What would be a better choice.
Also I found that media evaporated rather quickly in the chamber making media ingredients more concentrated. Any advice regarding that issue?
Thank you very much for any advise.
Olga
Olga Makarova, PhD
Research Specialist
5323 MSRB III
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