Localization of beta-D-galactosidase activity in semithin epon sections of embryonic tissues using differential interference contrast optics.
In the present study we describe a method for the histochemical demonstration of beta-D-galactosidase activity on tissue sections processed for light microscopy at high resolution. 5-Bromo-indolyl-beta-D-galactopyranoside (Bluo-Gal) was utilized as an indigogenic method for the demonstration of Escherichia coli beta-D-galactosidase reporter gene activity whose expression was studied in a transgenic line where the enzyme, with a nuclear localization signal (nlacZ), is under the transcriptional control of a striated muscle-specific promoter. At the light-microscopic level, by using Differential Interference Contrast (DIC) optics, the reaction product was detected as precipitates in the form of fine birefringent crystals. These were located around and inside the nuclei of beta-gal-expressing cells. This simple method allows an easy and rapid identification of few or even one labeled cell(s) within large microscopic fields (whole embryos) and the labeled cell(s) can be evaluated both morphologically and quantitatively.
Poggi
P
P
Institute of Human Normal Anatomy, University of Pavia, Italy.
Icaro-Cornaglia
A
A
Coletta
M
M
Kelly
R
R
Cossu
G
G
D000818:Animals; D004622:Embryo, Mammalian / Q000201:enzymology*; D004853:Epoxy Resins; D006651:Histocytochemistry / Q000379:methods; D051379:Mice; D008822:Mice, Transgenic; D008857:Microscopy, Interference / Q000379:methods; D008867:Microtomy / Q000379:methods; D016613:Plastic Embedding / Q000379:methods; D001616:beta-Galactosidase / Q000378:metabolism*
D004853:Epoxy Resins; C004875:EPON; D001616:beta-Galactosidase