Abstract
Apart from the spatial resolution enhancement, scaling of temporal resolution, equivalently the imaging throughput, of fluorescence microscopy is of equal importance in advancing cell biology and clinical diagnostics. Yet, this attribute has mostly been overlooked because of the inherent speed limitation of existing imaging strategies. To address the challenge, we employ an all-optical laser-scanning mechanism, enabled by an array of reconfigurable spatiotemporally-encoded virtual sources, to demonstrate ultrafast fluorescence microscopy at line-scan rate as high as 8 MHz. We show that this technique enables high-throughput single-cell microfluidic fluorescence imaging at 75,000 cells/second and high-speed cellular 2D dynamical imaging at 3,000 frames per second, outperforming the state-of-the-art high-speed cameras and the gold-standard laser scanning strategies. Together with its wide compatibility to the existing imaging modalities, this technology could empower new forms of high-throughput and high-speed biological fluorescence microscopy that was once challenged.
Highlights
Fluorescence microscopy is a versatile tool for scientific research and biomedical applications
Faster laser scanning up to hundreds of kHz can be achieved by acousto-optic deflectors (AODs) and electro-optic deflectors (EODs), these scanners suffer from the limited angular beam steering range (i.e. imaging field of view (FOV)) and the number of resolvable scanned points [6]
To highlight its versatile potential in the applications where high-speed and high-throughput measurements are critical, yet challenging, we present two demonstrations of free-space angular-chirp-enhanced delay (FACED) fluorescence imaging, namely highthroughput imaging flow cytometry at a fluorescence imaging throughput of 75,000 cells/second, and high-speed dynamical single-cell imaging at a 2D frame rate as high as 3,000 fps
Summary
Fluorescence microscopy is a versatile tool for scientific research and biomedical applications. The challenge stems from the inherent speed limitations in the common image capture schemes. The speed-versus-sensitivity trade-off in the typical image sensor arrays (e.g. charge-coupled device (CCD), or complementary metal-oxide semiconductor (CMOS) cameras) hinders fast dynamical fluorescence microscopy with a time resolution of millisecond or below, especially under the photon-budget-limited scenario [4]. Faster laser scanning up to hundreds of kHz can be achieved by acousto-optic deflectors (AODs) and electro-optic deflectors (EODs), these scanners suffer from the limited angular beam steering range (i.e. imaging field of view (FOV)) and the number of resolvable scanned points (and the resolution) [6]. AOD and EOD are intrinsically the dispersive elements which disperse (and distort) the broadband beams – making it non-trivial to be used in laser-scanning multi-photon imaging unless careful dispersion compensation is implemented [7]
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