Photophysics of Calcium Green 1 invitro and in live cells

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Publication Date
DOI 10.1039/A900584F
Impact Factor 3.676
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Abstract

The absolute measurement of calcium activity in cells is hindered by the presence of different forms of the probe (adsorbed, protonated). Time-resolved fluorescence spectroscopy can be a sensitive technique to separate contaminating forms of the probe. The fluorescent probe Calcium Green 1 was characterised, invitro, in terms of calcium and also pH dependence, albumin adsorption, extent of hydrolysis, viscosity and medium polarity. A 1:1 stoichiometry was obtained for the calcium complexation with a Kd value that agrees with reported values. A pKa of the BAPTA moiety was measured as 6.7. The fluorescence lifetimes are slightly dependent on polarity but not on viscosity. It was observed that most of the interfering forms of the probe share the common lifetimes of 0.5 and 3.5 ns. An alternative mechanism for Calcium Green 1 sensing is proposed based on a ground state conformational equilibrium affecting the photoinduced electron transfer.

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Physical Chemistry Chemical Physics

Physical Chemistry Chemical Physics
CiteScore: 5.5
Self-citation Rate: 10.3%
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Physical Chemistry Chemical Physics (PCCP) is an international journal co-owned by 19 physical chemistry and physics societies from around the world. This journal publishes original, cutting-edge research in physical chemistry, chemical physics and biophysical chemistry. To be suitable for publication in PCCP, articles must include significant innovation and/or insight into physical chemistry; this is the most important criterion that reviewers and Editors will judge against when evaluating submissions. The journal has a broad scope and welcomes contributions spanning experiment, theory, computation and data science. Topical coverage includes spectroscopy, dynamics, kinetics, statistical mechanics, thermodynamics, electrochemistry, catalysis, surface science, quantum mechanics, quantum computing and machine learning. Interdisciplinary research areas such as polymers and soft matter, materials, nanoscience, energy, surfaces/interfaces, and biophysical chemistry are welcomed if they demonstrate significant innovation and/or insight into physical chemistry. Joined experimental/theoretical studies are particularly appreciated when complementary and based on up-to-date approaches.

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