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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editor: Gregory W. Faris
  • Vol. 4, Iss. 9 — Sep. 4, 2009

Flow visualization and flow cytometry with holographic video microscopy

Fook Chiong Cheong, Bo Sun Rémi Dreyfus, Jesse Amato-Grill, Ke Xiao, Lisa Dixon, and David G. Grier  »View Author Affiliations


Optics Express, Vol. 17, Issue 15, pp. 13071-13079 (2009)
http://dx.doi.org/10.1364/OE.17.013071


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Abstract

The video stream captured by an in-line holographic microscope can be analyzed on a frame-by-frame basis to track individual colloidal particles’ three-dimensional motions with nanometer resolution, and simultaneously to measure their sizes and refractive indexes. Through a combination of hardware acceleration and software optimization, this analysis can be carried out in near real time with off-the-shelf instrumentation. An efficient particle identification algorithm automates initial position estimation with sufficient accuracy to enable unattended holographic tracking and characterization. This technique’s resolution for particle size is fine enough to detect molecular-scale coatings on the surfaces of colloidal spheres, without requiring staining or fluorescent labeling. We demonstrate this approach to label-free holographic flow cytometry by detecting the binding of avidin to biotinylated polystyrene spheres.

© 2009 Optical Society of America

OCIS Codes
(100.2960) Image processing : Image analysis
(120.7250) Instrumentation, measurement, and metrology : Velocimetry
(180.6900) Microscopy : Three-dimensional microscopy
(090.1995) Holography : Digital holography

ToC Category:
Instrumentation, Measurement, and Metrology

History
Original Manuscript: March 31, 2009
Revised Manuscript: July 7, 2009
Manuscript Accepted: July 11, 2009
Published: July 16, 2009

Virtual Issues
Vol. 4, Iss. 9 Virtual Journal for Biomedical Optics
July 17, 2009 Spotlight on Optics

Citation
Fook Chiong Cheong, Bo Sun Rémi Dreyfus, Jesse Amato-Grill, Ke Xiao, Lisa Dixon, and David G. Grier, "Flow visualization and flow cytometry with holographic video microscopy," Opt. Express 17, 13071-13079 (2009)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-17-15-13071


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