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Fluorescence linear dichroism imaging for quantifying membrane order.

Richard K P Benninger¹

¹Department of Bioengineering, University of Colorado, 1775 Aurora Court, Aurora, CO, 80045, USA, richard.benninger@ucdenver.edu.

Methods in Molecular Biology (Clifton, N.J.)

|October 22, 2014

Summary

This summary is machine-generated.

This study details fluorescent linear dichroism (fLD) imaging for measuring membrane order and topology in living cells. This advanced microscopy technique quantifies fluorophore orientation to reveal plasma membrane dynamics.

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Single-Molecule Imaging of Lateral Mobility and Ion Channel Activity in Lipid Bilayers using Total Internal Reflection Fluorescence TIRF Microscopy

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Published on: February 17, 2023

Area of Science:

Cell biology
Biophysics
Microscopy

Background:

Cellular plasma membranes are ordered, dynamic, and heterogeneous environments crucial for signaling.
Existing fluorescent microscopy methods offer high spatial and temporal resolution for membrane studies.

Purpose of the Study:

To describe detailed methods for performing fluorescent linear dichroism (fLD) imaging in biological membranes.
To enable spatially resolved measurement of membrane order and sub-resolution membrane topology.

Main Methods:

Utilizing a polarization-resolved fluorescence method, fluorescent linear dichroism (fLD) imaging.
Quantifying the orientation of membrane-bound fluorophores in living cells, specifically the plasma membrane.
Detailed protocols for sample preparation, microscopy system requirements, data collection, and image analysis.

Main Results:

Spatially resolved measurements of membrane order are achievable using fLD imaging.
Sub-resolution membrane topology, such as ruffling, can be detected and quantified.
The method is applicable to biological membrane environments like the plasma membrane of living cells.

Conclusions:

Fluorescent linear dichroism (fLD) imaging provides a powerful tool for analyzing membrane order and dynamics.
This technique offers high spatial and temporal resolution for studying cellular membrane organization.
Detailed methods are provided for the implementation and interpretation of fLD imaging in cell biology research.