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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: Oct 15, 2025

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules

Published on: August 8, 2019

Micromirror Total Internal Reflection Microscopy for High-Performance Single Particle Tracking at Interfaces.

Xuanhui Meng¹, Adar Sonn-Segev¹, Anne Schumacher¹

¹Physical and Theoretical Chemistry Laboratory, Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3TA, U.K.

|October 25, 2021

Summary

This summary is machine-generated.

This study introduces micromirror-based total internal reflection dark field microscopy for enhanced single particle tracking. It achieves nanometer precision at high speeds, ideal for observing nanoscale motion.

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Last Updated: Oct 15, 2025

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules

Published on: August 8, 2019

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

Area of Science:

Physical Sciences
Biophysics
Nanotechnology

Background:

Single particle tracking (SPT) is crucial for observing nanoscale motion in various scientific fields.
Fluorescence microscopy excels in high-background settings like cellular environments.
Scattering-based detection offers superior localization precision and speed due to an unlimited photon budget.

Purpose of the Study:

To develop a microscopy technique combining high background suppression with excellent localization precision and speed for SPT.
To demonstrate the capability of this new method for characterizing nanoscale dynamics at interfaces.

Main Methods:

Utilized micromirror-based total internal reflection dark field microscopy.
Employed 20 nm gold nanoparticles as probes.
Achieved 6 μs exposure times and a 25 × 25 μm² field of view.

Main Results:

Demonstrated background suppression comparable to interferometric scattering microscopy.
Achieved nanometer localization precision for 20 nm gold nanoparticles.
Characterized sub-nanometer deterministic flows of nanoparticles at liquid-liquid interfaces.

Conclusions:

Micromirror-based TIR-dark field microscopy offers a powerful new tool for high-performance SPT.
This technique approaches the optimal balance of background suppression, localization precision, and temporal resolution for nanoparticle tracking at interfaces.