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Total Internal Reflection Fluorescence Microscopy

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.

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Updated: Aug 20, 2025

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy VA-TIRFM

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy VA-TIRFM

Published on: October 2, 2012

Advanced quantification for single-cell adhesion by variable-angle TIRF nanoscopy.

Dalia El Arawi¹, Cyrille Vézy¹, Régis Déturche¹

¹Light, nanomaterials, nanotechnologies, ERL CNRS 7004, Université de Technologie de Troyes, Troyes, France.

Biophysical Reports

|November 25, 2022

Summary

This summary is machine-generated.

Variable-angle total internal reflection fluorescence (vaTIRF) nanoscopy quantifies cell adhesion from specific to nonspecific interactions. This method reveals that U87MG glioma cells overexpressing α5 integrins show higher contractility but lower adhesion energy.

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Last Updated: Aug 20, 2025

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy VA-TIRFM

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy VA-TIRFM

Published on: October 2, 2012

Imaging Molecular Adhesion in Cell Rolling by Adhesion Footprint Assay

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Published on: September 27, 2021

Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy

Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy

Published on: January 21, 2019

Area of Science:

Biophysics
Cell Biology
Nanotechnology

Background:

Existing cell adhesion techniques often overlook weak, nonspecific interactions.
Studying the full spectrum of cell adhesion requires advanced imaging methods.

Purpose of the Study:

To comprehensively investigate cell adhesion, from specific to nonspecific interactions.
To introduce and validate variable-angle total internal reflection fluorescence (vaTIRF) nanoscopy for cell adhesion studies.

Main Methods:

Utilized variable-angle total internal reflection fluorescence (vaTIRF) nanoscopy.
Mapped cell topography and cell cortex refractive index in real time.
Quantified cell-substrate binding energy and cell contractility.

Main Results:

vaTIRF successfully distinguished between high and low adhesive cell-substrate contacts.
Demonstrated a correlation between cell contractility and cell-substrate binding energy.
U87MG glioma cells overexpressing α5 integrins exhibited higher contractility and lower adhesion energy.

Conclusions:

vaTIRF nanoscopy is a powerful tool for quantitative analysis of cell adhesion dynamics.
Integrin expression levels influence cell contractility and adhesion energy.
Nonspecific cell-substrate interactions play a crucial role in overall cell adhesion.