Quantitative in situ detection of human β1-interferon mRNA

Feng-Jyu Tang-Nelson, Paul O.P. Ts'o, Stephen A. Lesko

Research output: Contribution to journalArticle

Abstract

A rapid, quantitative in situ mRNA hybridization method was developed to study human β1-interferon gene expression through the application of biotinylated DNA probes and computer-assisted microscopy. The β1-interferon DNA probe was chemically biotinylated and used for the hybridization to β1-interferon mRNA in fixed human HT1080 cells induced with poly (IC). For the purpose of quantitation, a computerized microphotometric system was employed for acquisition of signal intensity generated by streptavidin-alkaline phosphatase or streptavidin-FITC (fluorescein isothiocyanate), which were used to detect the hybrids formed after in situ hybridization. About 60% of the cells exhibited hybridization signals in induced cells. The speed, specificity and quantitation of this non-isotopic in situ hybridization method should be generally useful to measure gene expression at the single cell level.

Original languageEnglish (US)
Pages (from-to)41-44
Number of pages4
JournalActa Histochemica et Cytochemica
Volume26
Issue number1
DOIs
StatePublished - Jan 1 1993
Externally publishedYes

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Physiology
  • Biochemistry
  • Cell Biology

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