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      An expanded palette of genetically encoded Ca²⁺ indicators.

      Science (New York, N.Y.)

      Transfection, Spectrometry, Fluorescence, chemistry, Recombinant Fusion Proteins, Rats, Protein Engineering, metabolism, Neurons, Molecular Sequence Data, genetics, Luminescent Proteins, Humans, HeLa Cells, Green Fluorescent Proteins, Fluorescence Resonance Energy Transfer, Fluorescence, Directed Molecular Evolution, Calcium Signaling, analysis, Calcium, Caenorhabditis elegans, Animals, Genetically Modified, Animals, Adenosine Triphosphate

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          Abstract

          Engineered fluorescent protein (FP) chimeras that modulate their fluorescence in response to changes in calcium ion (Ca(2+)) concentration are powerful tools for visualizing intracellular signaling activity. However, despite a decade of availability, the palette of single FP-based Ca(2+) indicators has remained limited to a single green hue. We have expanded this palette by developing blue, improved green, and red intensiometric indicators, as well as an emission ratiometric indicator with an 11,000% ratio change. This series enables improved single-color Ca(2+) imaging in neurons and transgenic Caenorhabditis elegans. In HeLa cells, Ca(2+) was imaged in three subcellular compartments, and, in conjunction with a cyan FP-yellow FP-based indicator, Ca(2+) and adenosine 5'-triphosphate were simultaneously imaged. This palette of indicators paints the way to a colorful new era of Ca(2+) imaging.

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          Author and article information

          Journal
          21903779
          3560286
          10.1126/science.1208592

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