Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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.

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview

Attenuated total reflectance (ATR) infrared spectroscopy is a powerful analytical technique used to study the composition of materials. It is widely employed in chemistry, materials science, forensic science, and other fields where sample characterization is required. ATR has several advantages over traditional transmission IR spectroscopy, including the requirement of little to no sample preparation and the ability to analyze a wide range of samples.
The ATR process begins by directing a beam...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Quantifying 3D live-cell membrane dynamics using dynamic metal-induced energy transfer spectroscopy (dynaMIET).

Science advances·2026

Same author

Persistence diagrams as morphological signatures of cells: A method to measure and compare cells within a population.

PLoS computational biology·2026

Same author

Mapping Optical Chirality with Single Fluorescent Molecules.

Nano letters·2026

Same author

Versatile Microfluidics Platform for Enhanced Multitarget Super-Resolution Microscopy.

ACS nano·2026

Same author

The near-infrared bacteriophytochrome-derived fluorescent protein PENELOPE enables RESOLFT superresolution microscopy.

Proceedings of the National Academy of Sciences of the United States of America·2025

Same author

Self-regulating and self-oscillating metal-organic framework hybrid plasmonic metasurfaces.

Nature communications·2025

Same journal

Generalizable framework for multi-site bone density prediction using non-dominant wrist optical biomarkers.

Biomedical optics express·2026

Same journal

Erratum: Review of dynamic optical coherence tomography for intracellular motility [Invited]: errata.

Biomedical optics express·2026

Same journal

Digital-micromirror-device-based illumination strategies for background suppression in single-molecule localization microscopy.

Biomedical optics express·2026

Same journal

Synergistic combination of convective self-assembly and hollow core fiber for sensitive SERS detection of glucose molecules.

Biomedical optics express·2026

Same journal

Multimodal diagnostic network integrating infrared and mass spectra for lung cancer.

Biomedical optics express·2026

Same journal

Multimodal Optical Biosensing for Precision Medicine and Healthcare: Introduction to the feature issue.

Biomedical optics express·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Dec 23, 2025

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Published on: May 3, 2022

Metasurface-based total internal reflection microscopy.

Antu Nehuen Gortari¹, Sophie Bouchoule¹, Edmond Cambril¹

¹Ctr. de Nanosciences et de Nanotechnologies, Univ. Paris-Sud, Univ. Paris-Saclay, France.

Biomedical Optics Express

|April 29, 2020

Summary

This summary is machine-generated.

We developed a novel dielectric metasurface for total internal reflection fluorescence microscopy (TIRFM). This low-cost metasurface enables high-contrast bioimaging of cellular surfaces, offering an accessible alternative to traditional methods.

More Related Videos

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

Published on: November 16, 2019

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

Related Experiment Videos

Last Updated: Dec 23, 2025

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Published on: May 3, 2022

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

Published on: November 16, 2019

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

Area of Science:

Optics and Photonics
Materials Science
Biotechnology

Background:

Dielectric metasurfaces offer advanced control over light wavefronts for visible light applications.
Total Internal Reflection Fluorescence Microscopy (TIRFM) is crucial for high-contrast imaging of cell membranes and surface molecules.
Conventional objective-type TIRFM relies on expensive, bulky objectives and has a limited field-of-view.

Purpose of the Study:

To introduce a novel, cost-effective dielectric metasurface for TIRFM.
To demonstrate a metasurface-based approach as a user-friendly alternative to conventional TIRFM setups.
To enable high-resolution imaging of biological samples at the cellular surface.

Main Methods:

Fabrication of periodic 2D arrays of asymmetric TiO2 nanostructures on borosilicate glass.
Characterization of light coupling efficiency and angular distribution into the glass substrate.
Validation of the metasurface's performance in TIRFM by imaging immunostained human mesenchymal stem cells.

Main Results:

The metasurface efficiently couples up to 70% of incident light into the first diffraction order at 65° in glass, exceeding the critical angle for TIR.
Minimal light leakage (~7%) into propagating modes reduces background fluorescence.
Successful imaging of cellular structures over a 200 µm x 200 µm field-of-view was achieved.

Conclusions:

The developed dielectric metasurface provides a low-cost, easy-to-use platform for TIRFM.
This technology offers high contrast, low photodamage, and high surface selectivity for fluorescence imaging.
Metasurface-based TIRFM presents a valuable tool for advanced bioimaging applications.