| Autor: |
Nguyen, Cuong, Upadhyay, Hansini, Murphy, Michael, Borja, Gabriel, Rozsahegyi, Emily J., Barnett, Adam, Brookings, Ted, McManus, Owen B., Werley, Christopher A. |
| Zdroj: |
Biomedical Optics Express; February 2019, Vol. 10 Issue: 2 p789-806, 18p |
| Abstrakt: |
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. |
| Databáze: |
Supplemental Index |
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