Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes

  title={Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes},
  author={Thomas Thestrup and Julia Litzlbauer and Ingo Bartholom{\"a}us and Marsilius Mues and Luigi Russo and Hod Dana and Yuri Kovalchuk and Yajie Liang and Georgios Kalamakis and Yvonne Laukat and Stefan Becker and Gregor Witte and Anselm Geiger and Taylor Allen and Lawrence C Rome and Tsai-Wen Chen and Douglas S Kim and O. V. Garaschuk and Christian Griesinger and Oliver Griesbeck},
  journal={Nature Methods},
The quality of genetically encoded calcium indicators (GECIs) has improved dramatically in recent years, but high-performing ratiometric indicators are still rare. Here we describe a series of fluorescence resonance energy transfer (FRET)-based calcium biosensors with a reduced number of calcium binding sites per sensor. These 'Twitch' sensors are based on the C-terminal domain of Opsanus troponin C. Their FRET responses were optimized by a large-scale functional screen in bacterial colonies… CONTINUE READING
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