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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.
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...
Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
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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Related Experiment Video

Updated: Jun 4, 2026

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

Principles of Total Internal Reflection Microscopy (TIRFM).

David Zenisek, David Perrais

    CSH Protocols
    |March 2, 2011
    PubMed
    Summary

    Total internal reflection fluorescence microscopy (TIRFM) enables high-resolution imaging of cellular events near surfaces. This technique is valuable for studying exocytosis in neurons and other cell types.

    Area of Science:

    • Cell Biology
    • Microscopy Techniques
    • Neuroscience

    Background:

    • Total internal reflection fluorescence microscopy (TIRFM) is a specialized imaging method.
    • It excels at visualizing events at the cell-cell or cell-substrate interface.
    • TIRFM utilizes the evanescent wave generated at a high refractive index boundary.

    Purpose of the Study:

    • To explain the fundamental principles of TIRFM.
    • To provide guidance on establishing a TIRFM system.
    • To highlight TIRFM's applications in studying cellular processes.

    Main Methods:

    • Selective imaging of fluorescent molecules near a glass interface.
    • Application of TIRFM to observe single vesicle exocytosis.
    • Utilizing fluorescent dyes (FM1-43) and fluorescent proteins (NPY-EGFP).

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

    Published on: May 3, 2022

    Single Molecule Fluorescence Microscopy on Planar Supported Bilayers
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    Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

    Published on: October 31, 2015

    Related Experiment Videos

    Last Updated: Jun 4, 2026

    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

    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

    Single Molecule Fluorescence Microscopy on Planar Supported Bilayers
    20:00

    Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

    Published on: October 31, 2015

    Main Results:

    • Demonstrated TIRFM's capability in studying exocytosis of synaptic vesicles in neurons.
    • Showcased TIRFM's utility in analyzing dense core granule exocytosis in chromaffin cells.
    • Provided a foundational understanding of TIRFM setup and theory.

    Conclusions:

    • TIRFM is a powerful technique for near-surface cellular event visualization.
    • It offers high resolution for studying dynamic processes like exocytosis.
    • The article serves as a guide for implementing and understanding TIRFM.