Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Immunofluorescence Microscopy

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.

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Lenacapavir-induced lattice hyperstabilization is central to HIV-1 capsid failure at the nuclear pore complex and in the cytoplasm.

eLife·2026

Same author

Systematic discovery of pro- and anti-HIV host factors in primary human CD4+ T cells.

Cell·2026

Same author

HIV-1 uncoating location dictates sites of integration.

Nature communications·2026

Same author

Development of a Complementation Assay to Monitor Pan-Coronavirus 3C-like Protease Activity.

Viruses·2026

Same author

Elucidating the Mechanism by Which HIV-1 Nucleocapsid Mutations Confer Resistance to Integrase Strand Transfer Inhibitors.

bioRxiv : the preprint server for biology·2025

Same author

Lenacapavir-induced Lattice Hyperstabilization is Central to HIV-1 Capsid Failure at the Nuclear Pore Complex and in the Cytoplasm.

bioRxiv : the preprint server for biology·2025

Search research articles

Related Experiment Video

Updated: Jun 26, 2025

Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy

Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy

Published on: October 7, 2010

Single-Virion Analysis: A Method to Visualize HIV-1 Particle Content Using Fluorescence Microscopy.

Alice Duchon¹, Ryan C Burdick², Vinay K Pathak²

¹Viral Recombination Section, HIV Dynamics and Replication Program, National Cancer Institute, Frederick, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)

|May 14, 2024

Summary

Analyzing human immunodeficiency virus type 1 (HIV-1) virion content is crucial for understanding viral replication. A new single-virion analysis method uses fluorescence microscopy to determine the presence and distribution of molecules within individual HIV-1 particles.

Keywords:

Fluorescence microscopy Gag HIV particle imaging RNA genome packaging Virus assembly

More Related Videos

Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection

Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection

Published on: October 29, 2020

Detection of Viral RNA by Fluorescence in situ Hybridization FISH

Detection of Viral RNA by Fluorescence in situ Hybridization FISH

Published on: May 5, 2012

Related Experiment Videos

Last Updated: Jun 26, 2025

Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy

Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy

Published on: October 7, 2010

Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection

Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection

Published on: October 29, 2020

Detection of Viral RNA by Fluorescence in situ Hybridization FISH

Detection of Viral RNA by Fluorescence in situ Hybridization FISH

Published on: May 5, 2012

Area of Science:

Virology
Molecular Biology
Microscopy

Background:

HIV-1 virions assemble with viral RNA, cellular RNAs, and proteins.
Some virion components are essential for replication, while others, like host proteins, can inhibit it.
Traditional biochemical assays lack the resolution to determine molecule distribution within virion populations.

Purpose of the Study:

To introduce and validate a single-virion analysis method for studying HIV-1 particle composition.
To overcome the limitations of traditional biochemical assays in assessing virion content distribution.
To provide a flexible tool for analyzing nucleic acid and protein incorporation into HIV-1 particles.

Main Methods:

Development of a single-virion analysis technique utilizing fluorescence microscopy.
Labeling of HIV-1 unspliced RNA with a fluorescently tagged RNA-binding protein (Protein A).
Labeling of Gag proteins with a distinct fluorescent protein (Protein B) to identify viral particles.

Main Results:

The method allows for the direct visualization and quantification of molecules within individual HIV-1 particles.
Demonstrated the ability to determine the proportion of particles containing unspliced HIV-1 RNA.
Successfully adapted the method to study the incorporation of APOBEC3 proteins and modified Gag proteins into virions.

Conclusions:

Single-virion analysis is a powerful and flexible method for detailed investigation of HIV-1 virion content.
This technique provides insights into both the presence and distribution of viral and host molecules.
Facilitates a deeper understanding of HIV-1 assembly and replication dynamics.