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Related Concept Videos

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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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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Updated: Dec 18, 2025

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy
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Spatiotemporal Analysis of Caveolae Dynamics Using Total Internal Reflection Fluorescence Microscopy.

Yosuke Senju1, Shiro Suetsugu2

  • 1Research Institute for Interdisciplinary Science (RIIS), Okayama University, Okayama, Japan. yosuke.senju@okayama-u.ac.jp.

Methods in Molecular Biology (Clifton, N.J.)
|June 18, 2020
PubMed
Summary

Total internal reflection fluorescence microscopy (TIRFm) visualizes cellular events near the plasma membrane. This technique enhances signal-to-noise for studying caveolae dynamics and their physiological roles.

Keywords:
CaveolaeEndocytosisKymographLateral diffusionTotal internal reflection fluorescence microscopy

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Area of Science:

  • Cell Biology
  • Biophysics
  • Microscopy

Background:

  • Plasma membrane dynamics are crucial for cellular functions.
  • Caveolae are vital for membrane tension, endocytosis, and signaling.
  • Observing these events requires high-resolution imaging techniques.

Purpose of the Study:

  • To apply Total Internal Reflection Fluorescence Microscopy (TIRFm) for analyzing cellular events at the plasma membrane.
  • To investigate the dynamics of caveolae using TIRFm.

Main Methods:

  • Utilizing Total Internal Reflection Fluorescence Microscopy (TIRFm).
  • Focusing illumination on molecules near the glass-cell interface.
  • Minimizing background fluorescence for enhanced signal detection.

Main Results:

  • TIRFm provides a high signal-to-noise ratio for plasma membrane observation.
  • The method allows detailed analysis of molecular localization and dynamics.
  • Specific application demonstrated for studying caveolae dynamics.

Conclusions:

  • TIRFm is a powerful tool for studying plasma membrane events.
  • The technique offers superior visualization of cellular dynamics near the membrane.
  • This approach facilitates research into the physiological roles of structures like caveolae.