Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Herpes simplex virus-cell interactions studied by immunogold cryosection electron microscopy.

Helle Lone Jensen1, Bodil Norrild

  • 1Department of Pathology, Rigshospitalet, University Hospital of Copenhagen, Copenhagen, Denmark.

Methods in Molecular Biology (Clifton, N.J.)
|October 28, 2004
PubMed
Summary

This study presents a reproducible cryosection electron microscopy technique for high-resolution immunolocalization of multiple viral antigens. The method enhances visualization of virus-cell interactions and preserves ultrastructure.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Celebrating a century of APMIS: a legacy of pathology, microbiology, and immunology.

APMIS : acta pathologica, microbiologica, et immunologica Scandinavica·2024
Same author

Meta-analysis of penile cancer: conceptual interpretations - Authors' reply.

The Lancet. Oncology·2019
Same author

Prevalence of human papillomavirus DNA and p16<sup>INK4a</sup> in penile cancer and penile intraepithelial neoplasia: a systematic review and meta-analysis.

The Lancet. Oncology·2018
Same author

Human papillomavirus and p16 in squamous cell carcinoma and intraepithelial neoplasia of the vagina.

International journal of cancer·2018
Same author

Heading for centennial anniversary and beyond.

APMIS : acta pathologica, microbiologica, et immunologica Scandinavica·2017
Same author

Correlation between HPV status at T and N sites of oropharyngeal squamous cell carcinomas.

Acta oto-laryngologica·2017

Area of Science:

  • Cell Biology
  • Virology
  • Microscopy

Background:

  • High-resolution imaging of virus-cell interactions is crucial for understanding viral pathogenesis.
  • Existing immunolabeling techniques can have limitations in sensitivity and specificity.

Purpose of the Study:

  • To develop and optimize a reliable postembedding immunogold labeling technique for cryosection electron microscopy.
  • To enable high-resolution, multi-antigen detection in a single ultrathin cryosection.

Main Methods:

  • A three-layer indirect immunogold labeling protocol for ultrathin cryosections.
  • Utilized primary monoclonal antibodies from the same species for simultaneous antigen detection.
  • Employed a buffered paraformaldehyde-glutaraldehyde fixative to block non-specific antibody binding sites.

Related Experiment Videos

Main Results:

  • Successfully demonstrated co-localization of herpes simplex virus type 1 glycoproteins (gC-1 and gD-1) in nuclear membranes.
  • Visualized labeled viral particles in the perinuclear space and virions within the endoplasmic reticulum and Golgi apparatus.
  • The method preserved both ultrastructure and antigenicity, avoiding issues like variable particle size and cross-labeling.

Conclusions:

  • The optimized indirect immunogold labeling technique is simple, reproducible, and suitable for studying virus-cell interactions.
  • This method offers superior immunolabeling compared to two-layer methods and preserves ultrastructural integrity.
  • Effective blocking of secondary antibody binding sites ensures specific and reliable multi-antigen detection.