Title | A near-infrared genetically encoded calcium indicator for in vivo imaging. |
Publication Type | Journal Article |
Year of Publication | 2021 |
Authors | AA Shemetov, MV Monakhov, Q Zhang, JE Canton-Josh, M Kumar, M Chen, ME Matlashov, X Li, W Yang, L Nie, DM Shcherbakova, Y Kozorovitskiy, J Yao, N Ji, and VV Verkhusha |
Journal | Nat Biotechnol |
Volume | 39 |
Issue | 3 |
Start Page | 368 |
Pagination | 368 - 377 |
Date Published | 03/2021 |
Abstract | While calcium imaging has become a mainstay of modern neuroscience, the spectral properties of current fluorescent calcium indicators limit deep-tissue imaging as well as simultaneous use with other probes. Using two monomeric near-infrared (NIR) fluorescent proteins (FPs), we engineered an NIR Förster resonance energy transfer (FRET)-based genetically encoded calcium indicator (iGECI). iGECI exhibits high levels of brightness and photostability and an increase up to 600% in the fluorescence response to calcium. In dissociated neurons, iGECI detects spontaneous neuronal activity and electrically and optogenetically induced firing. We validated the performance of iGECI up to a depth of almost 400 µm in acute brain slices using one-photon light-sheet imaging. Applying hybrid photoacoustic and fluorescence microscopy, we simultaneously monitored neuronal and hemodynamic activities in the mouse brain through an intact skull, with resolutions of ~3 μm (lateral) and ~25-50 μm (axial). Using two-photon imaging, we detected evoked and spontaneous neuronal activity in the mouse visual cortex, with fluorescence changes of up to 25%. iGECI allows biosensors and optogenetic actuators to be multiplexed without spectral crosstalk. |
DOI | 10.1038/s41587-020-0710-1 |
Short Title | Nat Biotechnol |