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Live-cell biosensors based on the fluorescence lifetime of environment-sensing dyes.

Brian P Mehl | Pothiappan Vairaprakash | Li Li | Elizabeth Hinde | Christopher J MacNevin | Chia-Wen Hsu | Enrico Gratton | Bei Liu | Klaus M Hahn
Cell reports methods | 2024

In this work, we examine the use of environment-sensitive fluorescent dyes in fluorescence lifetime imaging microscopy (FLIM) biosensors. We screened merocyanine dyes to find an optimal combination of environment-induced lifetime changes, photostability, and brightness at wavelengths suitable for live-cell imaging. FLIM was used to monitor a biosensor reporting conformational changes of endogenous Cdc42 in living cells. The ability to quantify activity using phasor analysis of a single fluorophore (e.g., rather than ratio imaging) eliminated potential artifacts. We leveraged these properties to determine specific concentrations of activated Cdc42 across the cell.

Pubmed ID: 38503289

Associated grants

  • Agency: NIGMS NIH HHS, United States
    Id: R35 GM122596

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MATLAB (tool)

RRID:SCR_001622

Multi paradigm numerical computing environment and fourth generation programming language developed by MathWorks. Allows matrix manipulations, plotting of functions and data, implementation of algorithms, creation of user interfaces, and interfacing with programs written in other languages, including C, C++, Java, Fortran and Python. Used to explore and visualize ideas and collaborate across disciplines including signal and image processing, communications, control systems, and computational finance.

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