Hi, I was wondering if anyone working with gfp in bacterial cells has
experienced the problem which is driving me insane at present.
I previously had bacterial cells which expressed the gfp construct very
strongly and the cells were highly fluorescent when examined
microscopically, exciting them at the appropriate wavelength. However the
SAME cultures seem to have lost a substantial amount of the fluorescence and
from visibly seeing >90% of cells fluorescing I can now see <10%, although I
can still see the nonfluorescent cells. I have tried an activity dye which
seems to indicate that all the cells are viable so was wondering if anyone
has any idea how to overcome this problem.
Thanks in advance, Dr Karen P Scott.