Are you mounting your coverslips on your immunfluorescence slides
with a bleach retardant solution? We have been using Citifluor for
several years and it works very well. But, there are numerous other
alternatives. Also, it sounds like your fluorescence light source
needs to be adjusted to give you a uniform signal. Opitimally, the
entire field should be uniformly bright, so that you don't have dark
edges.
>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>First, I'd like to thank everyone for the advice on getting my yeast
cells to
stick to the slide. I've had some success. The problem now is
bleaching. My
cells stain, I can see them in the periphery of the field of view,
but as
soon as I bring them into the center for a good view, they VERY
RAPIDLY
bleach. My secondary ab is DTAF conjugated (same excitation &
emission
wavelengths as FITC). Is this bleaching a result of unstable
secondary? my
staining technique? or the UV source? Any references or info would be
greatly
appreciated.
Thanks in advance!
