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Nat Methods. 2015 Sep;12(9):852-8. doi: 10.1038/nmeth.3480. Epub 2015 Jul 13.

Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging.

Author information

1
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.
2
Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, California, USA.
3
Department of Molecular and Cellular Biology, University of California, Davis, Davis, California, USA.
4
Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.

Abstract

Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca(2+) indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca(2+) detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP).

PMID:
26167640
PMCID:
PMC4597790
DOI:
10.1038/nmeth.3480
[Indexed for MEDLINE]
Free PMC Article

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