Simultaneous voltage and calcium imaging and optogenetic stimulation with high sensitivity and a wide field of view
Simultaneous voltage and calcium imaging and optogenetic stimulation with high sensitivity and a wide field of view
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DOI：
10.1364/boe.10.000789
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发表时间：
2019-02-01
影响因子：
3.4
通讯作者：
Werley, Christopher A.
中科院分区：
文献类型：
--
作者：
Cuong Nguyen;Upadhyay, Hansini;Werley, Christopher A.
Transmembrane voltage and intracellular calcium concentration are coupled parameters essential to the function of neurons, cardiomyocytes. and other excitable cells. Here we introduce the Firefly-IIR microscope for simultaneous optogenetic stimulation and voltage and calcium imaging with fluorescent proteins using three spectrally distinct visible color bands. Firefly-HR combines patterned stimulation, near-total internal reflection laser excitation through a prism located between the sample and a water-immersion objective, and concurrent imaging of three color channels. The microscope has efficient light collection, low fluorescent background, and a large field of view (0.24 x 1.2 mm @1000 frames/sec). We characterize optical crosstalk and demonstrate capabilities with three applications: (1) probing synaptically connected neuronal microcircuits, (2) examining the coupling between neuronal action potentials and calcium influx, and (3) studying the pharmacology of paced human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) via simultaneous recordings of voltage, calcium, and contraction. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
