Localization of adenovirus DNA by in situ hybridization electron microscopy

Renjie Jiao, Wendou Yu, Mingxiao Ding, Zhonghe Zhai

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Biotinylated deoxyadenosine triphosphate (dATP) (Bio7dATP) and 3H deoxythymidine triphosphate (dTTP) labeled adenovirus DNA were hybridized in situ to thin sections of Lowicryl K4Membedded and wholemount 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 goldconjugated secondary antibodies and then criticalpoint dried while 3HdTTP labeled probe by electron microscopic autoradiography. On Lowicryl K4M sections, gold particles and silver grains were mainly restricted in the nucleus. Furthermore, wholemount results suggested that replicating adenovirus DNA is localized on the nuclear matrix of its host cell. In this paper, the described nonradioactive procedures for hybrid detection offered several advantages: (a) rapid signal detection; (ob) superior morphological preservation and spatial resolution; (c) precise localization; and (d) on Lowicryl K4M sections, signal to noise equivalent to radiolabeling. Copyright1992 WileyLiss, Inc.

Original languageEnglish (US)
Pages (from-to)23-31
Number of pages9
JournalMicroscopy Research and Technique
Volume21
Issue number1
DOIs
StatePublished - Mar 1 1992
Externally publishedYes

Keywords

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

ASJC Scopus subject areas

  • Anatomy
  • Histology
  • Instrumentation
  • Medical Laboratory Technology

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