Hi
I am wanting to take advantage of the high sensitivity of
phycoerythrins as flurophores for microscopic visualisation of
antibody probes in cell cultures.
I get a wonderful initial image but it fades to nothing in a few seconds.
I have tried using various antifadants, and the best give me a view-time
of 30s. It seems that the presence of glycerol in the mountant may
increase the bleaching rate. Calbiochem's FluorSave and ICN's
ImmunoFlour mountants seem best. The presence of DABCO dramatically
increases the bleach rate to less than 10 seconds. Vectorsheild
attenuates phycoerythrin fluoresence from the start as well as failing to
stabilise it. Home-made MOWIOL mountants with glycerol are worse than
the commercial mountants. Clearly, none of these are adequate for
microscopy.
Does anyone know of an antifadant for phycoerythrins which could
prolongue their fluorecence for a useable view-time? Or am I wasting time
here?
Thanks in advance for any replies
Julian Dye
Dept of Anatomy and Cell Biology
Imperial College School of Medicine
London
