TY - JOUR
T1 - Sensitive and rapid detection of β-galactosidase expression in intact cells by microinjection of fluorescent substrate
AU - Brustugun, Odd Terje
AU - Mellgren, Gunnar
AU - Gjertsen, Bjørn Tore
AU - Bjerkvig, Rolf
AU - Døskeland, Stein Ove
PY - 1995/8
Y1 - 1995/8
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0029155352&partnerID=8YFLogxK
U2 - 10.1006/excr.1995.1241
DO - 10.1006/excr.1995.1241
M3 - Article
C2 - 7641787
AN - SCOPUS:0029155352
SN - 0014-4827
VL - 219
SP - 372
EP - 378
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -