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Proceedings Paper

Short infrared laser pulses block action potentials in neurons
Author(s): Alex J. Walsh; Gleb P. Tolstykh; Stacey L. Martens; Bennett L. Ibey; Hope T. Beier
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Paper Abstract

Short infrared laser pulses have many physiological effects on cells including the ability to stimulate action potentials in neurons. Here we show that short infrared laser pulses can also reversibly block action potentials. Primary rat hippocampal neurons were transfected with the Optopatch2 plasmid, which contains both a blue-light activated channel rhodopsin (CheRiff) and a red-light fluorescent membrane voltage reporter (QuasAr2). This optogenetic platform allows robust stimulation and recording of action potential activity in neurons in a non-contact, low noise manner. For all experiments, QuasAr2 was imaged continuously on a wide-field fluorescent microscope using a Krypton laser (647 nm) as the excitation source and an EMCCD camera operating at 1000 Hz to collect emitted fluorescence. A co-aligned Argon laser (488 nm, 5 ms at 10Hz) provided activation light for CheRiff. A 200 mm fiber delivered infrared light locally to the target neuron. Reversible action potential block in neurons was observed following a short infrared laser pulse (0.26-0.96 J/cm2; 1.37-5.01 ms; 1869 nm), with the block persisting for more than 1 s with exposures greater than 0.69 J/cm2. Action potential block was sustained for 30 s with the short infrared laser pulsed at 1-7 Hz. Full recovery of neuronal activity was observed 5-30s post-infrared exposure. These results indicate that optogenetics provides a robust platform for the study of action potential block and that short infrared laser pulses can be used for non-contact, reversible action potential block.

Paper Details

Date Published: 8 February 2017
PDF: 7 pages
Proc. SPIE 10052, Optogenetics and Optical Manipulation, 100520J (8 February 2017); doi: 10.1117/12.2249519
Show Author Affiliations
Alex J. Walsh, National Research Council (United States)
Air Force Research Lab. (United States)
Gleb P. Tolstykh, General Dynamics Information Technology (United States)
Stacey L. Martens, Air Force Research Lab. (United States)
Bennett L. Ibey, Air Force Research Lab. (United States)
Hope T. Beier, Air Force Research Lab. (United States)


Published in SPIE Proceedings Vol. 10052:
Optogenetics and Optical Manipulation
Samarendra K. Mohanty; Nitish V. Thakor; E. Duco Jansen, Editor(s)

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