Localization of adenovirus DNA by in situ hybridization electron microscopy

R. Jiao, Wendou Yu, M. Ding, Z. H. Zhai

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Biotinylated deoxyadenosine triphosphate (dATP) (Bio-7-dATP) and 3H deoxythymidine triphosphate (dTTP) labeled adenovirus DNA were hybridized in situ to thin sections of Lowicryl K4M-embedded and whole-mount extracted HeLa cells infected with adenovirus. The biotinylated probe was detected by exposing the extracted cells or sections to antibodies against biotin followed by colloidal gold-conjugated secondary antibodies and then critical- point dried while 3H-dTTP labeled probe by electron microscopic autoradiography. On Lowicryl K4M sections, gold particles and silver grains were mainly restricted in the nucleus. Furthermore, whole-mount results suggested that replicating adenovirus DNA is localized on the nuclear matrix of its host cell. In this paper, the described non-radioactive procedures for hybrid detection offered several advantages: a) rapid signal detection; b) superior morphological preservation and spatial resolution; c) precise localization; and d) on Lowicryl K4M sections, signal to noise equivalent to radiolabeling.

Original languageEnglish (US)
Pages (from-to)23-31
Number of pages9
JournalMicroscopy Research and Technique
Volume21
Issue number1
DOIs
StatePublished - 1992
Externally publishedYes

Fingerprint

adenoviruses
Adenoviridae
Antibodies
in situ hybridization
Electron microscopy
In Situ Hybridization
electron microscopy
Electron Microscopy
DNA
deoxyribonucleic acid
Gold
gold
Signal detection
antibodies
nuclear matrix
Nuclear Matrix
Gold Colloid
radiolabeling
Silver
cells

Keywords

  • Autoradiography
  • Biotination
  • DNA replication
  • Nick translation
  • Nuclear matrix

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Anatomy
  • Instrumentation

Cite this

Localization of adenovirus DNA by in situ hybridization electron microscopy. / Jiao, R.; Yu, Wendou; Ding, M.; Zhai, Z. H.

In: Microscopy Research and Technique, Vol. 21, No. 1, 1992, p. 23-31.

Research output: Contribution to journalArticle

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