Intracellular dye heterogeneity determined by fluorescence lifetimes

Abstract

The cellular localization of a fluorescent probe molecule depends on both the chemical structure of the dye and the cellular environment. To study the number and types of environments in an epithelial cell line, we have measured in Madin-Darby canine kidney (MDCK) cells the fluorescence lifetimes of three structurally distinct fluorescent dyes--rhodamine-B, 3,3'-dihexadecylindocarbocyanine-(C3) (diI), and Collarein-incorporated into these cells. The latter is a rhodamine-cardiolipin conjugate that we designed and synthesized for the property of exclusive localization in the plasma membrane. The former two dyes required at least two exponential components to fit their fluorescence decay curves, while the decay of Collarein was characterized by a single exponential. These data are consistent with fluorescence microscopic observations, in which diI and rhodamine-B exhibit heterogeneous spatial distributions, while Collarein appears to be located on the cell surface.