WHaloCaMP data

Raw data for figures present in the manuscript linked in Related Materials. Each tab of the spreadsheet corresponds to a main figure panel in the linked manuscript. Details of the methodology used to acquire the data are described in the methods section of the manuscript linked in Related Materials. The manuscript, "A modular chemigenetic calcium indicator enables in vivo functional imaging with near-infrared light", describes engineering and validation of the chemigenetic calcium indicator WHaloCaMP, which can be used for sensing calcium ion concentration changes in the neurons of model organisms using fluorescence microscopy.Abstract: Genetically encoded fluorescent calcium indicators have revolutionized neuroscience and other biological fields by allowing cellular-resolution recording of physiology during behavior. However, we currently lack bright, genetically targetable indicators in the near infrared that can be used in animals. Here, we describe WHaloCaMP, a modular chemigenetic calcium indicator built from bright dye-ligands and protein sensor domains that can be genetically targeted to specific cell populations. 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 dye-ligands that efficiently label the central nervous system in animals. When bound to a near-infrared dye-ligand, WHaloCaMP1a is more than twice as bright as jGCaMP8s, and shows a 7× increase in fluorescence intensity and a 2.1 ns increase in fluorescence lifetime upon calcium binding. We use WHaloCaMP1a with near-infrared fluorescence emission to image Ca²⁺ responses in flies and mice, to perform three-color multiplexed functional imaging of hundreds of neurons and astrocytes in zebrafish larvae, and to quantitate calcium concentration using fluorescence lifetime imaging microscopy (FLIM).本数据集为「相关材料」中链接的手稿内配图的原始数据。电子表格的每个工作表对应链接手稿中的一幅主图子面板。数据采集方法的详细说明见「相关材料」中链接手稿的方法学章节。题为《模块化化学遗传钙指示剂可实现近红外光介导的体内功能成像》的手稿，阐述了化学遗传钙指示剂WHaloCaMP的构建与验证工作，该指示剂可通过荧光显微镜术检测模式生物神经元内的钙离子浓度变化。摘要：基因编码荧光钙指示剂（genetically encoded fluorescent calcium indicators）通过实现行为过程中生理活动的细胞分辨率记录，彻底革新了神经科学及其他生物领域。然而目前仍缺乏可用于动物实验的高亮度、可遗传靶向的近红外波段钙指示剂。本研究报道了WHaloCaMP——一种由高亮度染料配体与蛋白质传感器结构域构建而成的模块化化学遗传钙指示剂，可通过遗传靶向定位至特定细胞群。WHaloCaMP的荧光变化源于结合的染料被精准排布的色氨酸所介导的可逆淬灭。WHaloCaMP可兼容从绿光到近红外波段的罗丹明类染料配体，其中多款配体可高效标记动物体内的中枢神经系统。当结合近红外染料配体时，WHaloCaMP1a的亮度是jGCaMP8s的两倍以上，且在结合钙离子时荧光强度提升7倍、荧光寿命延长2.1纳秒。本研究使用搭载近红外荧光发射通道的WHaloCaMP1a，对果蝇与小鼠体内的钙离子响应进行成像，对斑马鱼幼体中数百个神经元与星形胶质细胞开展三色多路复用功能成像，并通过荧光寿命成像显微镜（fluorescence lifetime imaging microscopy, FLIM）定量检测钙离子浓度。