Design and Initial Characterization of a Small Near-Infrared Fluorescent Calcium Indicator

Near-infrared (NIR) genetically encoded calcium indicators (GECIs) are becoming powerful tools for neuroscience. Because of their spectral characteristics, the use of NIR GECIs helps to avoid signal loss from the absorption by body pigments, light-scattering, and autofluorescence in mammalian tissues. In addition, NIR GECIs do not suffer from cross-excitation artifacts when used with common fluorescent indicators and optogenetics actuators. Although several NIR GECIs have been developed, there is no NIR GECI currently available that would combine the high brightness in cells and photostability with small size and fast response kinetics. Here, we report a small FRET-based NIR fluorescent calcium indicator iGECInano. We characterize iGECInano in vitro, in non-neuronal mammalian cells, and primary mouse neurons. iGECInano demonstrates the improvement in the signal-to-noise ratio and response kinetics compared to other NIR GECIs.

Automated summary

Near-infrared (NIR) genetically encoded calcium indicators (GECIs) are powerful tools in neuroscience due to their spectral characteristics that minimize signal loss and cross-excitation artifacts. However, currently available NIR GECIs lack high brightness, photostability, small size, and fast response kinetics. In this study, we introduce a small FRET-based NIR fluorescent calcium indicator called iGECInano, which demonstrates improved signal-to-noise ratio and response kinetics compared to other NIR GECIs.