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<p>Genetically encoded calcium (Ca<sup>2+</sup>) indicators (GECIs) are essential tools for monitoring neuronal activity, but the performance of red fluorescent GECIs has remained limited. In particular, many red indicators are relatively dim, produce low signal-to-noise ratios and can undergo unwanted photoswitching when exposed to blue light, restricting their use in all-optical experiments that combine imaging with optogenetics or multicolor imaging. Here we show the development of PinkyCaMP, a Ca<sup>2+</sup> sensor based on the bright red fluorescent protein mScarlet. PinkyCaMP exhibits markedly improved brightness, photostability and signal-to-noise ratio compared to existing red GECIs, while remaining fully compatible with blue-light-based optogenetic and dual-color imaging approaches. PinkyCaMP is well-tolerated by neurons, showing no detectable toxicity or aggregation, both in vitro and in vivo. PinkyCaMP enables a broad spectrum of imaging modalities, including single-photon methods, such as fiber photometry, widefield imaging and miniature microscopy imaging, as well as two-photon imaging in awake mice.</p>

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PinkyCaMP: an mScarlet-based calcium sensor with enhanced brightness, photostability and multiplexing capabilities

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月24日

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  • Ryan Fink,
  • Shosei Imai,
  • Nala Gockel,
  • German Lauer,
  • Kim Renken,
  • Jonas Wietek,
  • Paul J. Lamothe-Molina,
  • Falko Fuhrmann,
  • Manuel Mittag,
  • Tim Ziebarth,
  • Annika Canziani,
  • Martin Kubitschke,
  • Vivien Kistmacher,
  • Anny Kretschmer,
  • Eva Sebastian,
  • Jana Ottens,
  • Dietmar Schmitz,
  • Takuya Terai,
  • Jan Gründemann,
  • Sami I. Hassan,
  • Tommaso Patriarchi,
  • Andreas Reiner,
  • Martin Fuhrmann,
  • Robert E. Campbell,
  • Olivia Andrea Masseck

摘要

Genetically encoded calcium (Ca2+) indicators (GECIs) are essential tools for monitoring neuronal activity, but the performance of red fluorescent GECIs has remained limited. In particular, many red indicators are relatively dim, produce low signal-to-noise ratios and can undergo unwanted photoswitching when exposed to blue light, restricting their use in all-optical experiments that combine imaging with optogenetics or multicolor imaging. Here we show the development of PinkyCaMP, a Ca2+ sensor based on the bright red fluorescent protein mScarlet. PinkyCaMP exhibits markedly improved brightness, photostability and signal-to-noise ratio compared to existing red GECIs, while remaining fully compatible with blue-light-based optogenetic and dual-color imaging approaches. PinkyCaMP is well-tolerated by neurons, showing no detectable toxicity or aggregation, both in vitro and in vivo. PinkyCaMP enables a broad spectrum of imaging modalities, including single-photon methods, such as fiber photometry, widefield imaging and miniature microscopy imaging, as well as two-photon imaging in awake mice.