Detection of LacZ‐Positive Cells in Living Tissue with Single‐Cell Resolution |
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| Authors: | Dr. Tomohiro Doura Dr. Mako Kamiya Dr. Fumiaki Obata Dr. Yoshifumi Yamaguchi Dr. Takeshi Y. Hiyama Takashi Matsuda Prof. Akiyoshi Fukamizu Prof. Masaharu Noda Prof. Masayuki Miura Prof. Yasuteru Urano |
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| Affiliation: | 1. Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan;2. PRESTO, Japan, Science and Technology Agency, Kawaguchi, Saitama, Japan;3. Graduate School of Pharmaceutical Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, Japan;4. Division of Molecular Neurobiology, National Institute for Basic Biology, Myodaiji-cho, Okazaki, Aichi, Japan;5. School of Life Science, The Graduate University for Advanced Studies, Myodaiji-cho, Okazaki, Aichi, Japan;6. Life Science Center, Tsukuba Advanced Research Alliance, Tsukuba, Ibaraki, Japan;7. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, 305–8577, Japan;8. CREST, Japan, Agency for Medical Research and Development, Chiyoda-ku, Tokyo, Japan |
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| Abstract: | The LacZ gene, which encodes Escherichia coli β‐galactosidase, is widely used as a marker for cells with targeted gene expression or disruption. However, it has been difficult to detect lacZ‐positive cells in living organisms or tissues at single‐cell resolution, limiting the utility of existing lacZ reporters. Herein we present a newly developed fluorogenic β‐galactosidase substrate suitable for labeling live cells in culture, as well as in living tissues. This precisely functionalized fluorescent probe exhibited dramatic activation of fluorescence upon reaction with the enzyme, remained inside cells by anchoring itself to intracellular proteins, and provided single‐cell resolution. Neurons labeled with this probe preserved spontaneous firing, which was enhanced by application of ligands of receptors expressed in the cells, suggesting that this probe would be applicable to investigate functions of targeted cells in living tissues and organisms. |
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| Keywords: | fluorescent probes lacZ quinone methide intermediates single-cell resolution β -galactosidase |
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