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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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Related Experiment Video

Updated: Jun 28, 2026

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
06:43

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 flow cytometry.

Jun Wang1, Ning Bao, Leela L Paris

  • 1Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, Indiana 47907, USA.

Analytical Chemistry
|November 15, 2008
PubMed
Summary
This summary is machine-generated.

Total internal reflection fluorescence flow cytometry (TIRF-FC) analyzes cell membranes at high throughput. This new method reveals cell population heterogeneity and subcellular protein localization with single-cell resolution.

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IP-FCM: Immunoprecipitation Detected by Flow Cytometry
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IP-FCM: Immunoprecipitation Detected by Flow Cytometry

Published on: December 2, 2010

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Last Updated: Jun 28, 2026

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06:43

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IP-FCM: Immunoprecipitation Detected by Flow Cytometry
12:17

IP-FCM: Immunoprecipitation Detected by Flow Cytometry

Published on: December 2, 2010

Area of Science:

  • Cell biology
  • Biophysics
  • Microscopy

Background:

  • Total internal reflection fluorescence microscopy (TIRFM) studies cell membrane events using evanescent waves.
  • TIRFM is limited by low throughput and inability to detect cell population heterogeneity.

Purpose of the Study:

  • Develop a high-throughput analytical tool for cell membrane analysis.
  • Overcome limitations of traditional TIRFM for population studies.

Main Methods:

  • Introduced total internal reflection fluorescence flow cytometry (TIRF-FC).
  • Utilized an elastomeric valve to direct cells to the evanescent field.
  • Achieved single-cell resolution at 100-150 cells/s throughput.

Main Results:

  • Demonstrated detection of subcellular fluorescent protein location differences.
  • Confirmed quantitative capabilities with data processing and analysis.
  • Showcased ability to analyze cell population dynamics near the cell surface.

Conclusions:

  • TIRF-FC offers high-throughput analysis of cell membrane events.
  • This method addresses limitations of TIRFM for population heterogeneity studies.
  • TIRF-FC is valuable for understanding cell surface dynamics and population-level events.