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Improved genetically encoded near-infrared fluorescent calcium ion indicators for in vivo imaging

By Yong Qian, 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, Robert E. Campbell

Posted 09 Apr 2020
bioRxiv DOI: 10.1101/2020.04.08.032433

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 two 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 C. 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. ### Competing Interest Statement The authors have declared no competing interest.

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