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Biomed Opt Express. 2016 Oct 20;7(11):4674-4684. eCollection 2016 Nov 1.

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

Author information

1
Department of Radiology and Biomedical Imaging, Yale University, New Haven, CT 06511, USA; Current affiliation: MGH/HST Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA 02129, USA; isencan@mgh.harvard.edu.
2
Biomedical Engineering, Yale University, New Haven, CT 06511, USA.
3
Department of Radiology and Biomedical Imaging, Yale University, New Haven, CT 06511, USA; Current affiliation: Department of Otology and Laryngology, Harvard Medical School, Massachusetts Eye and Ear Infirmary 243 Charles Street, Boston, MA 02114, USA.
4
Pediatrics, Yale University, New Haven, CT 06511, USA.
5
Applied Physics, Yale University, New Haven, CT 06511, USA.
6
Department of Radiology and Biomedical Imaging, Yale University, New Haven, CT 06511, USA; Biomedical Engineering, Yale University, New Haven, CT 06511, USA; Pediatrics, Yale University, New Haven, CT 06511, USA; Applied Physics, Yale University, New Haven, CT 06511, USA; michael.choma@yale.edu.

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

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