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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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Related Experiment Video

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In Situ Visualization of the Phase Behavior of Oil Samples Under Refinery Process Conditions

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High-Throughput In Situ Total Internal Reflection Imaging for Visualizing, Qualitatively Screening, and

Hao Lin^1,2, Zixin Wu¹, Qi Yang¹

¹Shenzhen Key Laboratory of Advanced Layered Materials for Value-Added Applications, Institute of Materials Research, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, P. R. China.

Analytical Chemistry

|February 27, 2026

Summary

This summary is machine-generated.

A new high-throughput platform using in situ total internal reflection imaging (TIRi) accelerates the discovery of efficient catalysts for hydrogen evolution. This method enables rapid screening and optimization of electrocatalysts for sustainable energy technologies.

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Area of Science:

Electrochemistry
Materials Science
Sustainable Energy

Background:

Advancing hydrogen energy is crucial for global energy transition and carbon neutrality.
Novel catalysts and in situ characterization are essential for efficient water electrolysis.
Current methods for catalyst evaluation can be time-consuming and lack scalability.

Purpose of the Study:

To develop a high-throughput in situ total internal reflection imaging (TIRi) platform for catalyst characterization.
To enable simultaneous visualization of catalyst uniformity, performance screening, and compositional analysis.
To accelerate the discovery and optimization of electrocatalysts for sustainable energy applications.

Main Methods:

Integration of a redesigned optical architecture with a 4 × 4 electrode array for in situ TIR imaging.
Utilizing optical contrast correlated with electrochemical response for catalyst evaluation.
Fabrication and testing of representative catalysts (Pt/C, NiFe, MoS2, WS2) and a Mo-Ru compositional-gradient alloy.

Main Results:

Verified spatial consistency and elucidated activity variations for hydrogen evolution reaction (HER) catalysts.
Identified an optimal Mo:Ru composition (1:0.339) for a Mo-Ru alloy catalyst.
Demonstrated the platform's ability to correlate electrocatalytic performance with compositional ratios.

Conclusions:

The developed TIRi platform offers a nondestructive, cost-efficient, and scalable method for high-throughput catalyst discovery.
The platform provides statistically robust results without compromising mechanistic insight.
This operando framework accelerates the rational design of electrocatalysts for sustainable energy technologies.