Line excitation array detection fluorescence microscopy at 0.8 million frames per second

Chris Martin, Tianqi Li, Evan Hegarty, Peisen Zhao, Sudip Mondal and Adela Ben-Yakar ()
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Chris Martin: The University of Texas at Austin
Tianqi Li: The University of Texas at Austin
Evan Hegarty: The University of Texas at Austin
Peisen Zhao: The University of Texas at Austin
Sudip Mondal: The University of Texas at Austin
Adela Ben-Yakar: The University of Texas at Austin

Nature Communications, 2018, vol. 9, issue 1, 1-10

Abstract: Abstract Three-dimensional, fluorescence imaging methods with ~1 MHz frame rates are needed for high-speed, blur-free flow cytometry and capturing volumetric neuronal activity. The frame rates of current imaging methods are limited to kHz by the photon budget, slow camera readout, and/or slow laser beam scanners. Here, we present line excitation array detection (LEAD) fluorescence microscopy, a high-speed imaging method capable of providing 0.8 million frames per second. The method performs 0.8 MHz line-scanning of an excitation laser beam using a chirped signal-driven longitudinal acousto-optic deflector to create a virtual light-sheet, and images the field-of-view with a linear photomultiplier tube array to generate a 66 × 14 pixel frame each scan cycle. We implement LEAD microscopy as a blur-free flow cytometer for Caenorhabditis elegans moving at 1 m s−1 with 3.5-µm resolution and signal-to-background ratios >200. Signal-to-noise measurements indicate future LEAD fluorescence microscopes can reach higher resolutions and pixels per frame without compromising frame rates.

Date: 2018
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DOI: 10.1038/s41467-018-06775-0

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