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

Thestrup, T., Litzlbauer, J., Bartholomaeus, I., Mues, M., Russo, L., Dana, H., et al. (2014). Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes. Nature Methods, 11(2), 175-182. doi: 10.1038/NMETH.2773.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0017-C32A-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-4829-5
Genre: Journal Article

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 Creators:
Thestrup, T., Author
Litzlbauer, J., Author
Bartholomaeus, I., Author
Mues, M., Author
Russo, L.1, Author              
Dana, H., Author
Kovalchuk, Y., Author
Liang, Y. J., Author
Kalamakis, G., Author
Laukat, Y., Author
Becker, S.1, Author              
Witte, G., Author
Geiger, A., Author
Allen, T., Author
Rome, L. C., Author
Chen, T. W., Author
Kim, D. S., Author
Garaschuk, O., Author
Griesinger, C.1, Author              
Griesbeck, O., Author
Affiliations:
1Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society, ou_578567              

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 Abstract: 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, refined by a secondary screen in rat hippocampal neuron cultures. We tested the in vivo performance of the most sensitive variants in the brain and lymph nodes of mice. The sensitivity of the Twitch sensors matched that of synthetic calcium dyes and allowed visualization of tonic action potential firing in neurons and high resolution functional tracking of T lymphocytes. Given their ratiometric readout, their brightness, large dynamic range and linear response properties, Twitch sensors represent versatile tools for neuroscience and immunology.

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Language(s): eng - English
 Dates: 2014-01-052014-02
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1038/NMETH.2773
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Title: Nature Methods
Source Genre: Journal
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Pages: - Volume / Issue: 11 (2) Sequence Number: - Start / End Page: 175 - 182 Identifier: -