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Sensitive and rapid detection of beta-galactosidase expression in intact cells by microinjection of fluorescent substrate.
- Source :
-
Experimental cell research [Exp Cell Res] 1995 Aug; Vol. 219 (2), pp. 372-8. - Publication Year :
- 1995
-
Abstract
- Bacterial beta-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-beta-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.
Details
- Language :
- English
- ISSN :
- 0014-4827
- Volume :
- 219
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Experimental cell research
- Publication Type :
- Academic Journal
- Accession number :
- 7641787
- Full Text :
- https://doi.org/10.1006/excr.1995.1241