We found Bodipy was a little better than FITC for co-labelling with Texas Red.
There is less bleed into the red filter with Bodipy. This using standard
Biorad 600 Ar-ion laser at 514nm.
When bleed can not be defeated, take advantage of the differntial bleaching
rate of Texas red and Bodipy/FITC. We published a graph of differntial
bleaching with a real double labelled specimaec in
Neuron, 5, pg 773-779, 1990. " Strategic location of calcium channels at transmitter
release sites of frog neuromuscular junction". The differential bleaching allowed us to
determine that bleed of the green signal through the red filter was not
a problem. Using zoom we bleached a small box then zoomed out and took another
picture. We could see the green bleached out in the box but intact elsewhere.
The Texas Red stain o Ca channels was not affected in the area where green
had been bleached out.
Milton Charlton
University of Toronto