Simultaneous recording of multiple cellular signaling events by frequency- and spectrally-tuned multiplexing of fluorescent probes.

ABSTRACT: Fluorescent probes that change their spectral properties upon binding to small biomolecules, ions, or changes in the membrane potential (V_m) are invaluable tools to study cellular signaling pathways. Here, we introduce a novel technique for simultaneous recording of multiple probes at millisecond time resolution: frequency- and spectrally-tuned multiplexing (FAST^M). Different from present multiplexing approaches, FAST^M uses phase-sensitive signal detection, which renders various combinations of common probes for V_m and ions accessible for multiplexing. Using kinetic stopped-flow fluorimetry, we show that FAST^M allows simultaneous recording of rapid changes in Ca²⁺, pH, Na⁺, and V_m with high sensi

SUBMITTER: Kierzek M 

PROVIDER: S-EPMC8700268 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature