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Related Experiment Video

Updated: May 10, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

High frame-rate fluorescence confocal angle-resolved linear dichroism microscopy.

Xiao Wang¹, Alla Kress, Sophie Brasselet

¹Aix-Marseille Université, Institut Fresnel, F-13013 Marseille, France.

The Review of Scientific Instruments

|June 8, 2013

Summary

This study introduces a faster angle-resolved linear dichroism technique for imaging molecular orientation in cells. The enhanced method achieves high-speed, precise orientation measurements using a spinning disk confocal system.

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Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

Angle-resolved linear dichroism (ARLD) provides molecular orientation information.
Existing ARLD techniques can be limited by acquisition speed.
High-speed imaging is crucial for studying dynamic cellular processes.

Purpose of the Study:

To develop a faster ARLD technique for molecular orientation imaging.
To combine spinning disk confocal microscopy with electro-optical polarization switching.
To quantify and compensate for polarization distortions and analyze signal-to-noise ratios.

Main Methods:

Implemented a spinning disk confocal excitation scheme.
Integrated electro-optical polarization switching and camera acquisition.
Quantified and compensated for polarization distortions introduced by the spinning disk.
Analyzed camera signal-to-noise characteristics for precision quantification.

Main Results:

Achieved precise molecular orientation imaging at full frame rates (approx. 1 second).
Successfully applied the technique to giant unilamellar vesicles and living cells.
Demonstrated imaging of lipid probes DiIC18 and di-8-ANEPPQ.

Conclusions:

The developed technique significantly boosts ARLD acquisition speed without compromising axial resolution.
This method enables rapid, precise molecular orientation mapping in biological samples.
The technique's performance is ultimately limited by fluorophore brightness and camera sensitivity.