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Cell Imaging Using Total Internal Reflection Fluorescence Microscopy02:51

Cell Imaging Using Total Internal Reflection Fluorescence Microscopy

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Source: Daniele, F., et. al TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis. J. Vis. Exp. (2015).This video demonstrates the imaging of human neuroblastoma cells using a total internal reflection fluorescence (TIRF) microscope to visualize fluorophore-tagged synaptic vesicles. The cells are first focused in epifluorescence mode. Adjustments are then made to achieve total internal reflection,...
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Total Internal Reflection Fluorescence Microscopy01:05

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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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Source: Winkle, C. C., et al. Utilizing Combined Methodologies to Define the Role of Plasma Membrane Delivery During Axon Branching and Neuronal Morphogenesis. J. Vis. Exp. (2016)This video demonstrates total internal reflection fluorescence (TIRF) microscopy for visualizing exocytic events in cortical neurons. A pH-sensitive green fluorescent protein (GFP) marker fluoresces upon vesicle fusion, with selective TIRF excitation capturing membrane-proximal exocytosis in time-lapse...
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Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins06:43

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

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We present a protocol for implementing interference-reflection microscopy and total-internal-reflection-fluorescence microscopy for the simultaneous imaging of dynamic microtubules and fluorescently labeled microtubule-associated...
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Related Experiment Video

Updated: Jan 19, 2026

Cell Imaging Using Total Internal Reflection Fluorescence Microscopy
02:51

Cell Imaging Using Total Internal Reflection Fluorescence Microscopy

Published on: June 17, 2025

603

Highly angular resolving beam separator based on total internal reflection.

Moritz Mihm, Ortwin Hellmig, André Wenzlawski

    Applied Optics
    |September 11, 2019
    PubMed
    Summary

    This study introduces an optical element that separates beams differing only in angle using total internal reflection. This innovation can reduce laser system size and complexity by shortening beam paths.

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    Last Updated: Jan 19, 2026

    Cell Imaging Using Total Internal Reflection Fluorescence Microscopy
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    Total Internal Reflection Fluorescence Microscopy for Visualization of Exocytic Events
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    Area of Science:

    • Optics and Photonics
    • Laser Systems Engineering

    Background:

    • Separating superimposed optical beams with slight angular differences is crucial for various laser applications.
    • Existing methods can be complex and lead to bulky optical setups.

    Purpose of the Study:

    • To develop a novel optical element for efficient angular separation of superimposed beams.
    • To demonstrate the element's effectiveness in a practical application involving acousto-optic modulators.

    Main Methods:

    • Utilizing total internal reflection at an air gap between two prisms to achieve angular resolution.
    • Designing an optical element adaptable to different wavelengths and component sizes.

    Main Results:

    • Successfully demonstrated the separation of superimposed beams with different diffraction orders at 800 nm.
    • The optical element effectively resolves and separates beams based on their angular divergence.

    Conclusions:

    • The presented optical element offers a compact and efficient solution for beam separation based on angle.
    • This technology has the potential to significantly reduce the size and complexity of laser systems.