entire membrane fluor vs capped - equally bright by FACS?
rhester at jaguar1.usouthal.edu
Wed Sep 12 12:08:24 EDT 2007
An investigator has fluorescence microscopy data showing very bright,
capped/polar fluorescence (present even at 4 C or with inclusion of
azide in the FACS buffer) and yet the same cells (the investigator says
they are 'polarized' cells) when examined by FACS are quite dim. The
FACS is a VantageSE and the fluorochrome is r-PE. Can anyone tell us
whether this should be the case or should a FACS detect capped
fluorescence just as efficiently as it does the same 'amount' of
fluorescnce spread out over the entire cell membrane?
Certainly, 'bright' and 'dim' are relative terms, especially when you
are comparing 'bright' fluorescence acquired with a fluorescence
microscope to 'dim' fluorescence acquired with the FACS, so perhaps this
should be treated as a theoretical question. Also, fluorescence area
has been collected to date on these samples; should they be acquiring,
and comparing, fluorescence height as well?
Thanks for any help.
Univ of South Alabama
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