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Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy

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Tutorial: fluorescence lifetime microscopy of membrane mechanosensitive Flipper probes.

Chloé Roffay¹, Juan Manuel García-Arcos¹, Pierrik Chapuis¹

¹Department of Biochemistry, University of Geneva, Geneva, Switzerland.

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|August 29, 2024

Summary

Researchers developed Flipper probes, fluorescent sensors that measure cellular forces by detecting changes in membrane tension. These probes offer a new way to visualize and quantify mechanical forces within cells, tissues, and organs.

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Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell

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Published on: August 4, 2015

Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Measuring mechanical forces within living cells is a significant technical hurdle in biological research.
Understanding cellular forces is crucial for various biological processes, including cell division, migration, and tissue development.

Purpose of the Study:

To introduce and detail the development of hydrophobic mechanosensing fluorescent probes, termed Flippers.
To provide a comprehensive guideline for utilizing Flipper probes with fluorescence lifetime imaging microscopy (FLIM) to measure cellular membrane biophysical parameters.

Main Methods:

Development of Flipper probes, which are hydrophobic fluorescent molecules whose fluorescence lifetime is sensitive to lipid packing.
Technical optimization of Flipper probes for effective cellular imaging.
Characterization of Flipper probes in diverse biological and in vitro systems using fluorescence lifetime imaging microscopy.

Main Results:

Flipper probes demonstrate a measurable change in fluorescence lifetime correlating with alterations in membrane tension.
Successful application of Flipper probes in various biological contexts, including cells, tissues, and organs.
Demonstrated ability of Flipper probes to report on long-range forces within biological systems.

Conclusions:

Flipper probes represent a valuable tool for quantifying membrane tension and cellular forces.
Fluorescence lifetime imaging microscopy with Flipper probes provides a robust method for assessing membrane biophysical parameters.
This technology enables new avenues for studying mechanobiology at cellular, tissue, and organ levels.