Ultrahigh-speed, phase-sensitive full-field interferometric confocal microscopy for quantitative microscale physiology

Biomed Opt Express. 2016 Oct 20;7(11):4674-4684. doi: 10.1364/BOE.7.004674. eCollection 2016 Nov 1.

Abstract

We developed ultra-high-speed, phase-sensitive, full-field reflection interferometric confocal microscopy (FFICM) for the quantitative characterization of in vivo microscale biological motions and flows. We demonstrated 2D frame rates in excess of 1 kHz and pixel throughput rates up to 125 MHz. These fast FFICM frame rates were enabled by the use of a low spatial coherence, high-power laser source. Specifically, we used a dense vertical cavity surface emitting laser (VCSEL) array that synthesized low spatial coherence light through a large number of narrowband, mutually-incoherent emitters. Off-axis interferometry enabled single-shot acquisition of the complex-valued interferometric signal. We characterized the system performance (~2 μm lateral resolution, ~8 μm axial gating depth) with a well-known target. We also demonstrated the use of this highly parallelized confocal microscopy platform for visualization and quantification of cilia-driven surface flows and cilia beat frequency in an important animal model (Xenopus embryos) with >1 kHz frame rate. Such frame rates are needed to see large changes in local flow velocity over small distance (high shear flow), in this case, local flow around a single ciliated cell. More generally, our results are an important demonstration of low-spatial coherence, high-power lasers in high-performance, quantitative biomedical imaging.

Keywords: (110.4980) Partial coherence in imaging; (170.1790) Confocal microscopy; (170.3880) Medical and biological imaging; (170.5380) Physiology; (180.3170) Interference microscopy; (250.7260) Vertical cavity surface emitting lasers.