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Updated: Jun 19, 2026

Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis

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Published on: November 6, 2019

Total internal reflection fluorescence (TIRF) microscopy.

Kenneth N Fish¹

¹Department of Psychiatry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.

Current Protocols in Cytometry

|October 10, 2009

Summary

This summary is machine-generated.

Total internal reflection fluorescence microscopy (TIRFM) uses an evanescent wave to excite fluorophores near surfaces. This technique is ideal for studying neuronal plasma membrane receptors with high optical sectioning.

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Area of Science:

Biophysics
Optical Microscopy
Cell Biology

Background:

Total internal reflection fluorescence microscopy (TIRFM) is an optical technique.
It excites fluorophores within a thin axial region (optical section) using an evanescent wave.
The evanescent wave is generated at the solid-liquid interface during total internal reflection.

Purpose of the Study:

To review the history, optical theory, and hardware configurations of TIRFM.
To provide experimental details and methodological considerations for TIRFM.
To facilitate the study of receptors at the plasma membrane in neurons.

Main Methods:

Utilizes total internal reflection of excitation light at a solid-liquid interface.
Generates an evanescent electromagnetic field that decays exponentially with distance.
Efficiently excites fluorescent molecules within a few hundred nanometers of the surface.

Main Results:

TIRFM provides high axial resolution by selectively exciting fluorophores near the interface.
The evanescent wave's properties enable precise optical sectioning.
The technique allows for detailed investigation of surface phenomena.

Conclusions:

TIRFM is a powerful tool for surface imaging and molecular studies.
Understanding TIRFM principles is crucial for its effective application.
This method is particularly valuable for neuroscience research on membrane receptors.