Simultaneous voltage and calcium imaging and optogenetic stimulation with high sensitivity and a wide field of view

Biomedical Optics Express
Cuong NguyenChristopher A Werley

Abstract

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-HR 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.

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Citations

Feb 20, 2020·Critical Reviews in Biochemistry and Molecular Biology·Xiang XuXiaofeng Jia
Sep 5, 2019·The Journal of Pharmacology and Experimental Therapeutics·Nathan P CoussensKurt Rasmussen
Dec 29, 2019·International Journal of Molecular Sciences·Alfons PenzkoferPeter Hegemann
Nov 10, 2020·Toxicological Sciences : an Official Journal of the Society of Toxicology·Ruth RobertsJennifer B Pierson
Dec 20, 2020·Nature Reviews. Cardiology·Emilia Entcheva, Matthew W Kay
Jun 25, 2021·Frontiers in Computational Neuroscience·Yuan ZengZhiyuan Yan
Jul 1, 2021·Journal of the American Chemical Society·Yuecheng ZhouBianxiao Cui
Aug 18, 2021·Biomaterials·Olaia F VilaGordana Vunjak-Novakovic

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