Imaging Membrane Proteins Using Total Internal Reflection Fluorescence Microscopy (TIRFM) in Mammalian Cells [PDF]
Bio-Protocol, 2023 The cell surfaceome is of vital importance across physiology, developmental biology, and disease states alike. The precise identification of proteins and their regulatory mechanisms at the cell membrane has been challenging and is typically determined ...
Kirin Gada +3 more
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Optimal Detection of Fusion Pore Dynamics Using Polarized Total Internal Reflection Fluorescence Microscopy [PDF]
Frontiers in Molecular Biosciences, 2021 The fusion pore is the initial narrow connection that forms between fusing membranes. During vesicular release of hormones or neurotransmitters, the nanometer-sized fusion pore may open-close repeatedly (flicker) before resealing or dilating irreversibly,
Joerg Nikolaus +12 more
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Quantifying F-actin patches in single melanoma cells using total-internal reflection fluorescence microscopy [PDF]
Scientific Reports, 2022 Total-internal reflection fluorescence (TIRF) microscope is a unique technique for selective excitation of only those fluorophore molecules in a cellular environment, which are located at the sub-diffraction axial distance of a cell’s contact-area ...
Elham Sheykhi +4 more
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A protocol for detecting elemental calcium signals (Ca2+ puffs) in mammalian cells using total internal reflection fluorescence microscopy [PDF]
STAR Protocols, 2021 Summary: This protocol outlines steps to visualize and detect Ca2+ puffs following photo-liberation of caged inositol-1,4,5-trisphosphate (IP3) from HEK-293 cells expressing only the native IP3R type 1 receptor using total internal reflection ...
Vikas Arige +4 more
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Single-Molecule Total Internal Reflection Fluorescence Microscopy. [PDF]
Cold Spring Harb Protoc, 2016 The advent of total internal reflection fluorescence (TIRF) microscopy has permitted visualization of biological events on an unprecedented scale: the single-molecule level. Using TIRF, it is now possible to view complex biological interactions such as cargo transport by a single molecular motor or DNA replication in real time.
Kudalkar EM, Davis TN, Asbury CL.
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Data Analysis for Total Internal Reflection Fluorescence Microscopy. [PDF]
Cold Spring Harb Protoc, 2016 In the microscopes we use to analyze total internal reflection fluorescence (TIRF), the emitted fluorescence is split chromatically, using dichroic filters, into either two or three different colors (“channels”). In our two-color instrument, the green emission wavelengths (405–488 nm; for imaging green fluorescent protein [GFP]-tagged proteins) and far-
Asbury CL.
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Preparation of Reactions for Imaging with Total Internal Reflection Fluorescence Microscopy. [PDF]
Cold Spring Harb Protoc, 2016 Here we present our standard protocol for studying the binding of kinetochore proteins to microtubules as a paradigm for designing single-molecule total internal reflection fluorescence (TIRF) microscopy experiments. Several aspects of this protocol require empirical optimization, including the method for anchoring the polymer or substrate to the ...
Kudalkar EM, Davis TN, Asbury CL.
europepmc +4 more sources
Communications Materials, 2021 Total internal reflection fluorescence microscopy typically relies on opaque optical waveguides, compatible only with upright microscopes. Here, a versatile approach is reported that uses CMOS-compatible transparent chips, demonstrated for the imaging of
Anish Priyadarshi +8 more
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Coverslip Cleaning and Functionalization for Total Internal Reflection Fluorescence Microscopy. [PDF]
Cold Spring Harb Protoc, 2016 Total internal reflection fluorescence (TIRF) microscopy allows visualization of biological events at the single-molecule level by restricting excitation to a precise focal plane near the coverslip and eliminating out-of-focus fluorescence. The quality of TIRF imaging relies on a high signal-to-noise ratio and therefore it is imperative to prevent ...
Kudalkar EM +3 more
europepmc +4 more sources
Imaging Early Steps of Sindbis Virus Infection by Total Internal Reflection Fluorescence Microscopy [PDF]
Advances in Virology, 2011 Sindbis virus (SINV) is an alphavirus that has a broad host range and has been widely used as a vector for recombinant gene transduction, DNA-based vaccine production, and oncolytic cancer therapy.
Youling Gu +2 more
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