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Related Experiment Video

Updated: Jan 23, 2026

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging

Published on: April 8, 2019

Build Your Own Microscope: Step-By-Step Guide for Building a Prism-Based TIRF Microscope.

Dalton R Gibbs¹, Anisa Kaur², Anoja Megalathan³

¹Department of Chemistry, Virginia Commonwealth University, 1001 West Main Street, Richmond, VA 23284, USA. gibbsdr@vcu.edu.

Methods and Protocols

|June 6, 2019

Summary

This study provides a detailed guide for building a prism-based total internal reflection fluorescence (pTIRF) microscope. This accessible method enables advanced single-molecule analysis for various life science research applications.

Keywords:

fluorescence microscope pTIRF single-molecule FRET single-molecule detection

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Last Updated: Jan 23, 2026

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Published on: April 8, 2019

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Area of Science:

Optics and Photonics
Biophysics
Materials Science

Background:

Prism-based total internal reflection fluorescence (pTIRF) microscopy is crucial for single-molecule analysis of biomolecules, nanostructures, and cells.
It utilizes an evanescent field for surface molecule excitation, enabling single-molecule detection, dynamics analysis, and kinetic data acquisition.
Currently, no commercial pTIRF microscopes are available, and building custom systems requires significant expertise.

Purpose of the Study:

To bridge the technology gap by providing a comprehensive, step-by-step guide for constructing a pTIRF microscope.
To enable researchers to build their own pTIRF systems for in vitro single-molecule imaging and nanostructure analysis.

Main Methods:

Detailed instructions for building a pTIRF microscope using commercially available components.
Characterization methods to validate the performance and capabilities of the built pTIRF microscope.

Main Results:

Successful construction and characterization of a functional pTIRF microscope.
Demonstration of the microscope's utility for in vitro single-molecule imaging and nanostructure analysis.

Conclusions:

The provided guide democratizes access to pTIRF technology for single-molecule research.
This DIY approach empowers researchers in life sciences and materials science to conduct advanced imaging experiments.