Hi all,
I did DAPI staining for MCF-7 cells as a control (not treated with anything at all). They are fixed cells and stained with DAPI for 10 mins and washed with PBS three times. Please see the attached pictures from a confocal microscope 1. under DAPI signal 2. brightfield 3. under GFP signal.
I don't understand why they give green fluorosence, do you think they are contaminated? There are also some black dots in the cytoplasm in the brightfield images. Could you please comment on this too?
Thank you
black dots in the cytoplasm in the brightfield images-- it "may" just the the characteristics of the cell , you can see the same dots in U2OS without contamination.
green fluorosence -- maybe a too long exposure time. By using a positive control (cell transefected with GFP), you could optimize the exposure condition.
Sena Karaosmanoglu ,
The bright field images look fine to me and I believe there's no contamination. Such signals are obtained when the laser power/ high-voltage/ gain are high. It would be clear if you mention those numbers for both DAPI and FITC channels. Also the media (if not PBS) in which cells are kept while imaging can result to such green emission.
Thanks
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