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Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

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.
Flow Cytometry01:23

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Updated: May 18, 2026

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)
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Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)

Published on: June 28, 2017

Optical microflow cytometer based on external total reflection.

Lung-Ming Fu1, Yao-Nan Wang

  • 1Department of Materials Engineering, National Pingtung University of Science and Technology, Pingtung, Taiwan.

Electrophoresis
|September 6, 2012
PubMed
Summary
This summary is machine-generated.

A new optical microflow cytometer simultaneously detects, counts, and sizes labeled and nonlabeled microparticles using external total reflection. This system accurately differentiates particle types in mixed samples for advanced microparticle analysis.

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Area of Science:

  • Biomedical Engineering
  • Optical Physics
  • Microfluidics

Background:

  • Accurate microparticle detection, enumeration, and sizing are crucial in various scientific fields.
  • Existing methods may face limitations in simultaneously analyzing labeled and nonlabeled particles.
  • Microfluidic devices offer miniaturized platforms for high-throughput analysis.

Purpose of the Study:

  • To propose and validate a novel optical microflow cytometer for simultaneous detection, enumeration, and sizing of labeled and nonlabeled microparticles.
  • To demonstrate the system's capability in distinguishing between fluorescent and nonfluorescent particles.
  • To enable accurate quantification of nonlabeled particles within mixed samples.

Main Methods:

  • Development of an optical microflow cytometer utilizing external total reflection.
  • Integration of a laser-induced fluorescence system, a polydimethylsiloxane (PDMS) chip, a plane mirror, and a dichroic beamsplitter.
  • Utilizing reflected light signals for total particle count/size and fluorescence signals for labeled particle detection.

Main Results:

  • The system successfully counted and sized fluorescent and nonfluorescent particles with diameters from 6 to 10.2 μm.
  • Demonstrated simultaneous detection and sizing of both labeled and nonlabeled microparticles.
  • Established a method to determine nonlabeled particle counts in mixed samples by signal subtraction.

Conclusions:

  • The proposed optical microflow cytometer provides a robust platform for simultaneous analysis of diverse microparticle populations.
  • External total reflection combined with fluorescence detection offers a sensitive and specific approach for microparticle characterization.
  • This technology has potential applications in diagnostics, environmental monitoring, and materials science requiring precise microparticle analysis.