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A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging
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A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging
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Journal Article
A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging
2024
Overview
Genetically encoded fluorescent calcium indicators allow cellular-resolution recording of physiology. However, bright, genetically targetable indicators that can be multiplexed with existing tools in vivo are needed for simultaneous imaging of multiple signals. Here we describe WHaloCaMP, a modular chemigenetic calcium indicator built from bright dye-ligands and protein sensor domains. Fluorescence change in WHaloCaMP results from reversible quenching of the bound dye via a strategically placed tryptophan. WHaloCaMP is compatible with rhodamine dye-ligands that fluoresce from green to near-infrared, including several that efficiently label the brain in animals. When bound to a near-infrared dye-ligand, WHaloCaMP shows a 7× increase in fluorescence intensity and a 2.1-ns increase in fluorescence lifetime upon calcium binding. We use WHaloCaMP1a to image Ca
2+
responses in vivo in flies and mice, to perform three-color multiplexed functional imaging of hundreds of neurons and astrocytes in zebrafish larvae and to quantify Ca
2+
concentration using fluorescence lifetime imaging microscopy (FLIM).
WHaloCaMP is a chemigenetic calcium indicator that can be combined with different rhodamine dyes for multiplexed or FLIM imaging in vivo, as demonstrated for calcium imaging in neuronal cultures, brain slices,
Drosophila
, zebrafish larvae and the mouse brain.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
