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

PLoS Biol. 2020 Nov 24;18(11):e3000965. doi: 10.1371/journal.pbio.3000965. eCollection 2020 Nov.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Brain / metabolism
  • Caenorhabditis elegans / metabolism
  • Calcium / metabolism*
  • Fluorescent Dyes
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • HeLa Cells
  • Humans
  • Indicators and Reagents
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Mice
  • Myocytes, Cardiac / metabolism
  • Neurons / metabolism
  • Optical Imaging / methods*
  • Optogenetics
  • Protein Engineering
  • Spectroscopy, Near-Infrared
  • Xenopus laevis / metabolism

Substances

  • Fluorescent Dyes
  • Indicators and Reagents
  • Luminescent Proteins
  • Green Fluorescent Proteins
  • Calcium