- Detection of LacZ-Positive Cells in Living Tissue with Single-Cell Resolution
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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.
- Doura, Tomohiro,Kamiya, Mako,Obata, Fumiaki,Yamaguchi, Yoshifumi,Hiyama, Takeshi Y.,Matsuda, Takashi,Fukamizu, Akiyoshi,Noda, Masaharu,Miura, Masayuki,Urano, Yasuteru
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- A mitochondria-targeted turn-on fluorescent probe based on a rhodol platform for the detection of copper(i)
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A new spirocyclized rhodol-based fluorescent probe has been developed for detecting mitochondrial Cu+. Alkylation of the hydroxy group of a xanthene moiety with a tris(2-pyridylmethyl)amine-based ligand induced the formation of a non-fluorescent spirocyclic structure. The reaction with Cu + in the presence of submillimolar concentrations of glutathione at physiological pH resulted in the elimination of the ligand together with an increase in the fluorescence of the rhodol fluorophore. This probe was used to visualize mitochondrial Cu+ in copper supplemented cells. This journal is the Partner Organisations 2014.
- Taki, Masayasu,Akaoka, Kazushi,Mitsui, Koji,Yamamoto, Yukio
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p. 4999 - 5005
(2014/07/07)
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- β-galactosidase fluorescence probe with improved cellular accumulation based on a spirocyclized rhodol scaffold
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We identified a rhodol bearing a hydroxymethyl group (HMDER) as a suitable scaffold for designing fluorescence probes for various hydrolases. HMDER shows strong fluorescence at physiological pH, but phenolic O-alkylation of HMDER results in a strong prefe
- Kamiya, Mako,Asanuma, Daisuke,Kuranaga, Erina,Takeishi, Asuka,Sakabe, Masayo,Miura, Masayuki,Nagano, Tetsuo,Urano, Yasuteru
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p. 12960 - 12963
(2011/10/05)
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