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Updated: May 25, 2026

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

Total internal reflection fluorescence microscopy imaging-guided confocal single-molecule fluorescence spectroscopy.

Desheng Zheng¹, Leonora Kaldaras, H Peter Lu

¹Department of Chemistry, Center for Photochemical Sciences, Bowling Green State University, Bowling Green, Ohio 43403, USA.

The Review of Scientific Instruments

|February 4, 2012

Summary

We developed a new spectroscopy system for analyzing molecules at interfaces. This integrated system allows for simultaneous sampling and detailed analysis of individual molecules, advancing studies in biological and chemical systems.

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Last Updated: May 25, 2026

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Area of Science:

Biophysics
Chemical Physics
Analytical Chemistry

Background:

Studying molecular dynamics at interfaces is crucial for understanding biological and chemical processes.
Existing techniques may lack the resolution or throughput for comprehensive interface analysis.

Purpose of the Study:

To develop an integrated spectroscopy system for analyzing two-dimensional interfaces.
To enable simultaneous sampling and in situ analysis of individual molecules at interfaces.

Main Methods:

Combined total internal reflection fluorescence microscopy imaging with confocal single-molecule fluorescence spectroscopy.
Utilized fluorescent microspheres for system calibration.
Performed single-molecule spectroscopy measurements.

Main Results:

Demonstrated successful calibration of the integrated system.
Successfully conducted single-molecule spectroscopy measurements at interfaces.
The system allows for simultaneous sampling of multiple molecules and in situ analysis of individual molecules.

Conclusions:

The integrated single-molecule spectroscopy system is a powerful tool for studying molecular dynamics.
This approach significantly enhances the study of interfaces in biological and chemical systems.