Red-shifted fluorogenic substrate for detecting lacZ-positive cells in living tissue with single-cell resolution.
Authors of this article are:
Ito H Kawamata Y Kamiya M Tsuda-Sakurai K Tanaka S Ueno T Komatsu T Hanaoka K Okabe S Miura M Urano Y.
A summary of the article is shown below:
The Escherichia coli lacZ gene encoding β-galactosidase is a widely used reporter, but few synthetic substrates are available for detecting its activity with single-cell resolution in living samples. Our recently reported fluorogenic substrate SPiDER-βGal is suitable for this purpose, but its hydrolysis product shows green fluorescence emission, and a red-shifted analogue is required for use in combination with green fluorescent protein (GFP) markers. Here, we describe the development of a red-shifted fluorogenic substrate for β-galactosidase, SPiDER-Red-βGal, based on silicon rhodol scaffold and by using a carboxylic group as the intramolecular nucleophile. LacZ-positive cells were successfully labeled with SPiDER-Red-βGal at the single cell resolution in living samples, and enabled us to visualize different cell types in combination with GFP markers.
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