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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.
Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...

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Visualization of Cortex Organization and Dynamics in Microorganisms, using Total Internal Reflection Fluorescence Microscopy
14:14

Visualization of Cortex Organization and Dynamics in Microorganisms, using Total Internal Reflection Fluorescence Microscopy

Published on: May 1, 2012

Imaging with total internal reflection fluorescence microscopy for the cell biologist.

Alexa L Mattheyses1, Sanford M Simon, Joshua Z Rappoport

  • 1Laboratory of Cellular Biophysics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.

Journal of Cell Science
|October 26, 2010
PubMed
Summary

Total internal reflection fluorescence (TIRF) microscopy selectively illuminates molecules near the cell membrane. This technique offers high-contrast imaging with reduced background and photodamage for cell biology research.

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Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biophysics

Background:

  • Total internal reflection fluorescence (TIRF) microscopy is a powerful technique for studying cellular events.
  • It selectively illuminates fluorophores within approximately 100 nm of the cell membrane.
  • This selective illumination reduces background noise and cellular photodamage.

Purpose of the Study:

  • To provide a comprehensive overview of Total internal reflection fluorescence (TIRF) microscopy.
  • To discuss its applications in cell biology.
  • To explain the physical principles, experimental setup, and troubleshooting for TIRF microscopy.

Main Methods:

  • Discussion of the physical principles behind TIRF microscopy.
  • Explanation of experimental setup considerations.
  • Troubleshooting common issues in TIRF experiments.

Main Results:

  • TIRF microscopy provides high-contrast images of the cell membrane.
  • It enables detailed analysis of molecular localization and dynamics near the plasma membrane.
  • Reduced background and photodamage allow for rapid imaging and sensitive detection.

Conclusions:

  • TIRF microscopy is a versatile tool for cell biology research.
  • Its ability to selectively image the cell periphery offers significant advantages.
  • Understanding its principles and application is crucial for researchers in the field.