[Histonet] dapi and GFP problem problem
Hey,
I have a GFP labeled g-protein coupled receptor that I am expressing
in HEK 293 cells. I cultured the cells on collagen coated glass
chamber slides, fixed the cells with 4% PFA, and then coverslipped
with Prolong Gold antifade containing dapi. Before I coverslipped I
checked the slide on a standard fluorescent microscope to make sure
everything was expressing correctly. My transduced cells had a nice
signal and my control was black. The day after coverslipping my
control cells showed a noticeable green signal, not quite a bright as
the GFP cells, but it is apparent. I took confocal pics and there
was clearly some green in my control cells.
Does anyone have a suggestions? I figure one of two things
happened. Either some cells slid around when I pressed down on the
coverslip or maybe I am getting some sort of bleedthrough. I am not
very good with the confocal yet, so I don't know if I did something
wrong here. Does anyone have a good protocol for growing 293 cells
on glass slides? I have heard there of some super adherent clones of
293 that I would like to try, although I haven't found a source for
them.
Any and all suggestions would be appreciated.
Thanks,
Caroline
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