Sensitive and rapid detection of β-galactosidase expression in intact cells by microinjection of fluorescent substrate

Odd Terje Brustugun, Gunnar Mellgren, Bjørn Tore Gjertsen, Rolf Bjerkvig, Stein Ove Døskeland*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

11 Citations (Scopus)

Abstract

Bacterial β-galactosidase, coded by lacZ, is a widely used reporter for studies of transcriptional activity of eukaryotic promoters at the single cell level. Unfortunately, current detection methods, like X-gal cytochemistry, are slow, have suboptimal sensitivity, and are incompatible with cell survival. By a novel approach based on microinjection into cells of the fluorogenic substrate 5-chloromethylfluorescein di-β-D-galactopyranoside lacZ gene expression was detected without affecting cell viability or proliferative capacity. The method was far more sensitive than the conventional X-gal cytochemistry in all cell systems tested (primary hepatocytes, fibroblasts, and glioma cells). Results were obtained within seconds to minutes after injection, and cells remained fluorescent for hours.

Original languageEnglish
Pages (from-to)372-378
Number of pages7
JournalExperimental Cell Research
Volume219
Issue number2
DOIs
Publication statusPublished - Aug 1995
Externally publishedYes

Fingerprint

Dive into the research topics of 'Sensitive and rapid detection of β-galactosidase expression in intact cells by microinjection of fluorescent substrate'. Together they form a unique fingerprint.

Cite this