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TitleImproved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging.
Publication TypeJournal Article
Year of Publication2020
AuthorsQian, Yong, Danielle M. Orozco Cosio, Kiryl D. Piatkevich, Sarah Aufmkolk, Wan-Chi Su, Orhan T. Celiker, Anne Schohl, Mitchell H. Murdock, Abhi Aggarwal, Yu-Fen Chang, Paul W. Wiseman, Edward S. Ruthazer, Edward S. Boyden, and Robert E. Campbell
JournalPLoS Biol
Volume18
Issue11
Paginatione3000965
Date Published2020 11
ISSN1545-7885
KeywordsAnimals, Brain, Caenorhabditis elegans, Calcium, Fluorescent Dyes, Green Fluorescent Proteins, HeLa Cells, Humans, Indicators and Reagents, Luminescent Proteins, Mice, Myocytes, Cardiac, Neurons, Optical Imaging, Optogenetics, Protein Engineering, Spectroscopy, Near-Infrared, Xenopus laevis
Abstract

Near-infrared (NIR) genetically encoded calcium ion (Ca2+) indicators (GECIs) can provide advantages over visible wavelength fluorescent GECIs in terms of reduced phototoxicity, minimal spectral cross talk with visible light excitable optogenetic tools and fluorescent probes, and decreased scattering and absorption in mammalian tissues. Our previously reported NIR GECI, NIR-GECO1, has these advantages but also has several disadvantages including lower brightness and limited fluorescence response compared to state-of-the-art visible wavelength GECIs, when used for imaging of neuronal activity. Here, we report 2 improved NIR GECI variants, designated NIR-GECO2 and NIR-GECO2G, derived from NIR-GECO1. We characterized the performance of the new NIR GECIs in cultured cells, acute mouse brain slices, and Caenorhabditis elegans and Xenopus laevis in vivo. Our results demonstrate that NIR-GECO2 and NIR-GECO2G provide substantial improvements over NIR-GECO1 for imaging of neuronal Ca2+ dynamics.

DOI10.1371/journal.pbio.3000965
Alternate JournalPLoS Biol
PubMed ID33232322
PubMed Central IDPMC7723245
Grant ListMOP-123514 / / CIHR / Canada
FS-154310 / / CIHR / Canada
R01 DA045549 / DA / NIDA NIH HHS / United States
R01 DA029639 / DA / NIDA NIH HHS / United States
/ HHMI / Howard Hughes Medical Institute / United States
FDN-143238 / / CIHR / Canada
U01 NS094246 / NS / NINDS NIH HHS / United States
RF1 NS113287 / NS / NINDS NIH HHS / United States
U01 NS090565 / NS / NINDS NIH HHS / United States
R01 MH122971 / MH / NIMH NIH HHS / United States