Affordable Access

In situ hybridization for light and electron microscopy: a comparison of methods for the localization of viral RNA using biotinylated DNA and RNA probes.

Authors
  • Troxler, M
  • Pasamontes, L
  • Egger, D
  • Bienz, K
Type
Published Article
Journal
Journal of Virological Methods
Publisher
Elsevier
Publication Date
Oct 01, 1990
Volume
30
Issue
1
Pages
1–14
Identifiers
PMID: 1964937
Source
Medline
License
Unknown

Abstract

We report procedures for in situ hybridization at the light and electron microscopic level for localization of viral RNA in poliovirus-infected, Lowicryl-embedded cells. We compare specificity and signal intensity of biotinylated, double-stranded DNA and single-stranded, strand-specific RNA probes, both corresponding to the same region of the poliovirus genome. The hybrids were detected with antibiotin antibodies or streptavidin with colloidal gold as a marker. Hybridization with the RNA probe was more sensitive and gave lower background than with DNA. Detection with immunogold proved to be by far more sensitive than with streptavidin. The hybridization and detection protocols for the DNA and the RNA probes could be applied without modification to light microscopic semi-thick sections as well as to electron microscopic ultrathin sections.

Report this publication

Statistics

Seen <100 times