Microscopy

Tom Nicholson tnicho1 at po-box.mcgill.ca
Mon Nov 4 17:43:11 EST 2002


Hello,

I was hoping someone could help me - I have made GFP constructs and
transfected them into my cells.  After sorting, the FACS means range
from about 80 -100 (with a negative of about 3-4).  However, when I take
these cells to the fluorescence microscope,  I do not see any
fluorescence.  I have been fixing my  cells with 4% formaldehyde,
followed by methanol treatment.  I then add a 1:1 solution of
PBS:glycerol, and seal with molten agarose.  I visualize using the FITC
filter on our microscope.

Any suggestions why I do not see anything would be greatly appreciated.

Thanks.




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