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Related Experiment Video

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

Ikbal Sencan1, Brendan K Huang2, Yong Bian3

  • 1Department 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.

Biomedical Optics Express
|November 30, 2016
PubMed
Summary

We developed ultra-high-speed microscopy to study tiny biological movements. This new technique uses a special laser to capture fast cellular flows and cilia beating in real-time.

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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Area of Science:

  • Biomedical Imaging
  • Microscopy Technology
  • Fluid Dynamics

Background:

  • Characterizing microscale biological motions and flows is crucial for understanding cellular functions.
  • Existing microscopy techniques often lack the speed and sensitivity required for dynamic, in vivo measurements.

Purpose of the Study:

  • To develop and demonstrate an ultra-high-speed, phase-sensitive, full-field reflection interferometric confocal microscopy (FFICM) system.
  • To enable quantitative characterization of in vivo microscale biological motions and flows at unprecedented speeds.

Main Methods:

  • Utilized a dense vertical cavity surface emitting laser (VCSEL) array to synthesize low spatial coherence, high-power light.
  • Employed off-axis interferometry for single-shot acquisition of complex-valued interferometric signals.
  • Achieved 2D frame rates exceeding 1 kHz and pixel throughput rates up to 125 MHz.

Main Results:

  • Demonstrated FFICM system performance with ~2 μm lateral resolution and ~8 μm axial gating depth.
  • Visualized and quantified cilia-driven surface flows and cilia beat frequency in Xenopus embryos at >1 kHz frame rate.
  • Showcased the system's ability to capture high shear flow dynamics around individual ciliated cells.

Conclusions:

  • Ultra-high-speed FFICM enables quantitative analysis of rapid biological processes previously unobservable.
  • Low spatial coherence, high-power lasers are key enablers for high-performance, quantitative biomedical imaging.
  • This technology offers significant potential for advancing research in developmental biology and cell motility.