Journal of Cell Science 1988-01-01

Coumarin-phalloidin: a new actin probe permitting triple immunofluorescence microscopy of the cytoskeleton.

J V Small, S Zobeley, G Rinnerthaler, H Faulstich

Index: J. Cell Sci. 89 , 21-24, (1988)

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Abstract

7-Diethylamino-3-(4-isothiocyanotophenyl)-4-methylcoumarin (CPITC) was coupled to amino-methyldithiolanophalloidin to produce a new phalloidin derivative, coumarin-phalloidin, fluorescent in the blue region of the spectrum. Coumarin-phalloidin binds to actin with around 100-fold less affinity than unconjugated phalloidin, but with enough avidity to make it a useful stain for actin filaments. Appropriate filter combinations permit triple immunofluorescence microscopy of the cytoskeleton with fluorescein and rhodamine conjugates together with coumarin-phalloidin.