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4 March 2019 Automated detection of action potentials in interferometric imaging of spiking cells (Conference Presentation)
Kevin C. Boyle, Tong Ling, Georges Goetz, Peng Zhou, Yi Quan, Felix S. Alfonso, Tiffany W. Huang, Daniel Palanker
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Proceedings Volume 10890, Label-free Biomedical Imaging and Sensing (LBIS) 2019; 1089025 (2019) https://doi.org/10.1117/12.2508871
Event: SPIE BiOS, 2019, San Francisco, California, United States
Abstract
Wide-field interferometric imaging systems can detect mechanical deformations of a cell during an action potential (AP), such as in quantitative phase microscopy, which is highly sensitive to the changing optical path length. This enables non-invasive optophysiology of spiking cells without exogeneous markers, but high-fidelity imaging of such deformations requires averaging of a large number of spikes synchronized by electrical recordings. We have developed new iterative methods for detecting single APs from quantitative phase microscopy of spiking cells, enabling an all-optical detection system with high accuracy and good temporal resolution. We demonstrate performance of the method across multiple preparations of spiking HEK-293 cells and compare the outcomes of the all-optical measurements with the ground truth detected on a multi-electrode array. We initially use a spike-triggered average, synchronized to an electrical recording, to measure deformations during the AP in spiking cells, which reach up to 3 nm (0.9 mrad) with a rise time of 4 ms and fall time of about 120 ms. Based on this knowledge of the AP dynamics, optical data analysis can provide reliable spike detection, within a standard deviation of 11.6 ms (9.7% of the length of the action potential) with an 8.5% false negative detection rate. The method is robust to natural variations between cells and can be modified to function without any prior knowledge of the AP dynamics. Such a system could achieve high-throughput measurements of network activity in culture and help identify the mechanisms linking cell deformations to the changes of transmembrane potential.
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Kevin C. Boyle, Tong Ling, Georges Goetz, Peng Zhou, Yi Quan, Felix S. Alfonso, Tiffany W. Huang, and Daniel Palanker "Automated detection of action potentials in interferometric imaging of spiking cells (Conference Presentation)", Proc. SPIE 10890, Label-free Biomedical Imaging and Sensing (LBIS) 2019, 1089025 (4 March 2019); https://doi.org/10.1117/12.2508871
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KEYWORDS
Action potentials
Interferometry
Microscopy
Data analysis
Imaging systems
Iterative methods
Temporal resolution
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