autofluorescence

Richard P Grant rgrant at see.sig.for.address
Thu Mar 27 07:08:59 EST 1997


Bill Kalionis (ogbk at gamgee.cc.flinders.edu.au) wrote:
} We are having problems with cells that autofluoresce brightly in the rhodamine 
} channel and to a lesser degree in the fluorescein channel. Does anybody have 

OK, not much help I know, but you could try switching to Texas Red, vs
rhodamine.  

Is it true autofluorescence - ie are the cells themselves
glowing without any fluorophore?  If so, then you could try using a
different mountant (maybe just changing the base buffer), or using the
infra red filter on your microscope (if it has one), changing the
excitation wavelength, or publish that your
cells really do glow ;)  Some cells - esp transformed cells - do seem to
AF naturally.  You might use DABCO in the mountant to quench free
radicals.

Of course, if you only see fluorescence on labelled cells, you've got a
different problem......


hth,
Richard

--
Richard P. Grant  MA  DPhil        University of Oxford
Nuffield Department Obstetrics and Gynaecology      FFPGP
rgrant at molbiol.ox.ac.uk            http://www.molbiol.ox.ac.uk/~rgrant
--------# "They're the shorter, squarer ones" - Will Carling #--------



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