Simultaneous detection of beta-galactosidase activity and surface antigen expression in viable haematopoietic cells.
The quantitation of intracellular beta-galactosidase activity has been described for viable cells. By using the fluorogenic substrate fluorescein-di-beta-D-galactopyranoside (FDG) in conjunction with flow cytometry, the proportion of positive cells as well as the level of expression can be determined. In this paper we describe beta-galactosidase expression in lymphoid and myeloid cells from transgenic mice that widely express beta-galactosidase from an inserted lacZ transgene. Both foetal and adult haematopoietic tissues are able to express beta-galactosidase. The intracellular fluorescence reflecting beta-galactosidase activity can be readily combined with fluorescently labelled antibodies against cell surface antigens. Thus, beta-galactosidase can be used as a marker in transplantation experiments to study the development of lymphoid and myeloid precursor cells.
Children's Medical Research Institute, Wentworthville, New South Wales, Australia.
C N
CN
Berger
S S
SS
Tan
K S
KS
Sturm
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis* / Q000097:blood; D015415:Biomarkers; D003433:Crosses, Genetic; D005260:Female; D005434:Flow Cytometry; D005452:Fluoresceins; D005697:Galactosides; D006412:Hematopoietic Stem Cells / Q000201:enzymology* / Q000276:immunology*; D008099:Liver / Q000196:embryology / Q000201:enzymology; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D013950:Thymus Gland / Q000196:embryology / Q000201:enzymology; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000378:metabolism*
D000954:Antigens, Surface; D015415:Biomarkers; D005452:Fluoresceins; D005697:Galactosides; C038848:fluorescein-digalactoside; D001616:beta-Galactosidase