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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.

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Related Experiment Video

Updated: Jun 4, 2026

Imaging Exocytosis in Retinal Bipolar Cells with TIRF Microscopy
11:54

Imaging Exocytosis in Retinal Bipolar Cells with TIRF Microscopy

Published on: June 9, 2009

Imaging Exocytosis with Total Internal Reflection Microscopy (TIRFM).

David Zenisek, David Perrais

    CSH Protocols
    |March 2, 2011
    PubMed
    Summary

    Total internal reflection fluorescence microscopy (TIRFM) directly images vesicles before fusion. This technique visualizes synaptic vesicles and dense core granules in neurons and chromaffin cells, offering new insights into exocytosis.

    Area of Science:

    • Cell biology
    • Neuroscience
    • Biophysics

    Background:

    • Electrophysiological methods indirectly infer pre-fusion events.
    • Total internal reflection fluorescence microscopy (TIRFM) excels at imaging cell-surface events.
    • TIRFM selectively visualizes fluorescent molecules near a high refractive index surface.

    Purpose of the Study:

    • To utilize TIRFM for directly observing vesicle fusion.
    • To investigate pre-fusion steps in exocytosis.
    • To study synaptic vesicles and dense core granules.

    Main Methods:

    • Application of Total Internal Reflection Fluorescence Microscopy (TIRFM).
    • Direct imaging of synaptic vesicles and dense core granules.
    • Study of exocytosis in retinal bipolar neurons and chromaffin cells.

    More Related Videos

    Imaging Plasma Membrane Deformations With pTIRFM
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    Imaging Plasma Membrane Deformations With pTIRFM

    Published on: April 2, 2014

    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

    Related Experiment Videos

    Last Updated: Jun 4, 2026

    Imaging Exocytosis in Retinal Bipolar Cells with TIRF Microscopy
    11:54

    Imaging Exocytosis in Retinal Bipolar Cells with TIRF Microscopy

    Published on: June 9, 2009

    Imaging Plasma Membrane Deformations With pTIRFM
    12:28

    Imaging Plasma Membrane Deformations With pTIRFM

    Published on: April 2, 2014

    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

    Main Results:

    • Direct visualization of vesicle behavior preceding fusion.
    • Detailed observation of synaptic vesicle and dense core granule dynamics.
    • Insights into the spatial and temporal aspects of exocytosis.

    Conclusions:

    • TIRFM provides direct visualization of pre-fusion vesicle dynamics.
    • This method enhances understanding of exocytosis mechanisms.
    • Applicable to various cell types involved in secretion.