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Related Concept Videos

Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...

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Visualizing HIV-1 Assembly at the T-Cell Plasma Membrane Using Single-Molecule Localization Microscopy.

Rayane Dibsy1, Kaushik Inamdar1, Cyril Favard1

  • 1CNRS, University of Montpellier, Institut de Recherche en Infectiologie de Montpellier - IRIM, UMR9004, Montpellier, France.

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Summary

Super-resolution microscopy visualizes human immunodeficiency virus type 1 (HIV-1) Gag protein assembly in real-time. This advanced technique allows detailed study of viral replication at the single-molecule level in host cells.

Keywords:
HIV-1PALMSTORMSingle-molecule light microscopyTIRF-MViral Gag protein assembly

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Area of Science:

  • Cell Biology
  • Virology
  • Microscopy

Background:

  • Optical fluorescence microscopy advancements enable visualization of nanoscale biological processes.
  • Super-resolution microscopy and fluorescent probes allow studying viral replication at the single-particle or protein level in living cells.

Purpose of the Study:

  • To present a protocol for visualizing human immunodeficiency virus type 1 (HIV-1) Gag assembly at the host T-cell plasma membrane.
  • To detail the use of super-resolution light microscopy for studying viral protein assembly.

Main Methods:

  • Utilizing Total Internal Reflection Fluorescence Microscopy (TIRF-M) combined with Single-Molecule Localization Microscopy (SMLM).
  • Describing TIRF equipment, T-cell culture for HIV-1, sample preparation for PALM and STORM, acquisition protocols, and Gag assembling cluster analysis.

Main Results:

  • TIRF-M and SMLM enable detection and characterization of viral protein assembly at the host cell plasma membrane.
  • Single protein-level analysis of HIV-1 Gag assembly is achievable.

Conclusions:

  • Super-resolution microscopy provides a powerful tool for investigating viral assembly dynamics.
  • The described protocol facilitates detailed studies of HIV-1 replication mechanisms at the molecular level.