Journal
/
Biology
/
Single-Cell Multiplexed Fluorescence Imaging to Visualize Viral Nucleic Acids and Proteins and Monitor HIV, HTLV, HBV, HCV, Zika Virus, and Influenza Infection
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
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

DOI:
10.3791/61843-v

•

07:24 min

•

October 29, 2020

•

, , , , ,
1Laboratory of Biochemical Pharmacology, Department of Pediatrics,Emory University School of Medicine, 2Department of Molecular Microbiology & Immunology,University of Missouri School of Medicine

Chapters

  • 00:04Introduction
  • 00:24Cell Seeding, Fixation and Permeabilization
  • 01:40Coverslip Immobilization and Protease Treatment
  • 02:31Target Probe and Amplifier Hybridization
  • 03:40Protein Immunostaining and Nuclear Counterstaining
  • 04:48Mounting
  • 05:28Results: Representative Single‐Cell Multiplexed Fluorescence Imaging
  • 06:54Conclusion

Summary

Automatic Translation

Automatic Translation

Presented here is a protocol for a fluorescence imaging approach, multiplex immunofluorescent cell-based detection of DNA, RNA, and protein (MICDDRP), a method capable of simultaneous fluorescence single-cell visualization of viral protein and nucleic acids of different type and strandedness. This approach can be applied to a diverse range of systems.

Tags

Keywords: Single-cellMultiplexedFluorescenceImagingViralNucleic AcidsProteinsHIVHTLVHBVHCVZikaInfluenzaImmunofluorescenceCell-based DetectionDNARNAProteinPoly-D-lysinePermeabilizationProteaseHybridizationMicroscopy

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/61843?article_type=v

Purification and Visualization of Influenza A Viral Ribonucleoprotein Complexes
check_url/61843?article_type=v

A Technique to Simultaneously Visualize Virus-Specific CD8+ T Cells and Virus-Infected Cells In situ
check_url/61843?article_type=v

NanoDrop Microvolume Quantitation of Nucleic Acids
check_url/61843?article_type=v

Concentration Determination of Nucleic Acids and Proteins Using the Micro-volume Bio-spec Nano Spectrophotometer
check_url/61843?article_type=v

Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
check_url/61843?article_type=v

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues
check_url/61843?article_type=v

Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method
check_url/61843?article_type=v

Functional Imaging of Viral Transcription Factories Using 3D Fluorescence Microscopy
check_url/61843?article_type=v

Single Cell Micro-aspiration as an Alternative Strategy to Fluorescence-activated Cell Sorting for Giant Virus Mixture Separation
check_url/61843?article_type=v

Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV

Read Article