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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.
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview01:13

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview

Attenuated total reflectance (ATR) infrared spectroscopy is a powerful analytical technique used to study the composition of materials. It is widely employed in chemistry, materials science, forensic science, and other fields where sample characterization is required. ATR has several advantages over traditional transmission IR spectroscopy, including the requirement of little to no sample preparation and the ability to analyze a wide range of samples.
The ATR process begins by directing a beam...
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Related Experiment Video

Updated: Jun 19, 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

Partial internal reflections on total internal reflection fluorescent microscopy.

Sanford M Simon1

  • 1Rockefeller University, New York, NY 10065, USA. simon@rockefeller.edu

Trends in Cell Biology
|October 13, 2009
PubMed
Summary
This summary is machine-generated.

Total internal reflection (TIR) fluorescence microscopy visualizes membrane events. This method requires careful validation to ensure observed signals accurately represent the biological phenomena under investigation.

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Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)
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Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)

Published on: October 2, 2012

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers
20:00

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Published on: October 31, 2015

Related Experiment Videos

Last Updated: Jun 19, 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

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)
09:14

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)

Published on: October 2, 2012

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers
20:00

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Published on: October 31, 2015

Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Fluorescence microscopy is vital for studying biological processes.
  • Visualizing narrow biological membranes (4nm) presents challenges due to light scattering.
  • Total internal reflection (TIR) fluorescence microscopy enhances membrane event visualization.

Purpose of the Study:

  • To address the challenge of validating signals in TIR fluorescence microscopy.
  • To ensure observed signals represent genuine biological phenomena.
  • To optimize the application of TIR microscopy for new biological problems.

Main Methods:

  • Development of techniques to narrow data collection planes.
  • Application of total internal reflection (TIR) principles.
  • Methodological validation strategies for TIR fluorescence microscopy.

Main Results:

  • TIR fluorescence microscopy offers improved visualization of membrane events.
  • Established methods exist to narrow the focal plane for enhanced imaging.
  • A critical need for signal validation in TIR microscopy was identified.

Conclusions:

  • TIR fluorescence microscopy is a powerful tool for membrane studies.
  • Careful validation is essential when applying TIR microscopy to new biological questions.
  • Further development in validation techniques will enhance TIR microscopy's utility.