Engineering a subcellular targetable, red-emitting, and ratiometric fluorescent probe for Ca2+ and its bioimaging applications |
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Authors: | Baocun Zhu Hongying Jia Xiaoling Zhang Yan Chen Haipeng Liu Weihong Tan |
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Institution: | (1) Department of Chemistry, School of Science, Beijing Institute of Technology, Beijing, 100081, China;(2) Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100080, China;(3) Center for Research at Bio/nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Shands Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, FL 32611-7200, USA; |
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Abstract: | A new, visible-light-excited and red-emitting fluorescent Ca2+ probe, STDBT, was synthesized, which consists of 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid as a Ca2+-chelating moiety and two benzothiazolium hemicyanine dyes as fluorophores. The spectral profiles of its free and Ca2+-bound forms were studied. Upon addition of Ca2+, the fluorescence spectra of STDBT displayed a significant enhancement (about 48-fold) in fluorescence intensity and a 20-nm
blueshift (from 600 to 580 nm) in the emission spectrum. Both the absorption and the excitation spectra of STDBT showed a
very large (more than 100 nm) hypsochromic shift in the long-wavelength maxima upon binding with Ca2+. Interestingly, in contrast with the commonly used Ca2+ indicator Fluo-3, when the acetoxymethyl ester of STDBT enters into cells, it distributes both in the cytosol and the nucleus,
but displays a very clear boundary between the two compartments. This allows STDBT to be used as a double targetable Ca2+ probe that can be used to report cytoplasmic Ca2+ and nuclear Ca2+ simultaneously. |
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