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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

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Flash-and-Freeze: A Novel Technique to Capture Membrane Dynamics with Electron Microscopy

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Published on: May 1, 2017

Mechanosensitive Conjugated Oligoelectrolytes for Visualizing Temporal Changes in Live Cells.

Ji-Yu Zhu^1,2, Samuel J W Chan^1,3, Hongyue Cui⁴

¹Department of Chemistry, National University of Singapore, Singapore, 117543, Singapore.

Angewandte Chemie (International Ed. in English)

|May 6, 2025

Summary

Researchers developed new fluorescent probes called conjugated oligoelectrolytes (COEs) that change color based on their environment. These probes can track changes in live cell membranes over time, offering insights into cellular mechanics.

Keywords:

Conjugated oligoelectrolytes Fluorescent Probes Fluorogenic Lipid Bilayer Mechanosensitivity

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Area of Science:

Biophysics
Cell Biology
Materials Science

Background:

Membrane-intercalating conjugated oligoelectrolytes (COEs) are fluorescent dyes used for bioimaging.
Current COEs visualize dye location but not dynamic cellular changes.
There is a need for probes that report on the physical properties of live cell membranes.

Purpose of the Study:

To develop novel, environmentally sensitive COEs for live-cell imaging.
To create probes capable of reporting temporal changes in intracellular membrane properties.
To utilize these probes for visualizing dynamic biophysical changes in cellular compartments.

Main Methods:

Design and synthesis of the COE-BY series featuring a BODIPY core and cationic pendant groups.
Characterization of photophysical properties, including two-photon absorption and NIR-II excitation.
Testing probe sensitivity to lipid composition and membrane tension in model vesicles and live cells.
Application of two-photon fluorescence lifetime imaging microscopy (2p-FLIM) for dynamic imaging.

Main Results:

The COE-BY series exhibits high two-photon absorption, NIR-II excitation, and brightness.
Probe emission lifetime is sensitive to lipid composition and membrane tension.
COE-BYPhOC4 successfully imaged changes in intracellular membrane mechanics.
COEs demonstrated stable vesicle-binding within the endolysosomal pathway for long-term monitoring.

Conclusions:

Environmentally sensitive COEs provide a new tool for visualizing temporal changes in live cell membrane physical properties.
These probes enable long-term monitoring of biophysical changes in intracellular vesicles.
The developed COEs advance bioimaging capabilities for studying cellular dynamics and mechanics.