Flow visualization and flow cytometry with holographic video microscopy

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

    Research output: Contribution to journalArticle

    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.

    Original languageEnglish (US)
    Pages (from-to)13071-13079
    Number of pages9
    JournalOptics Express
    Volume17
    Issue number15
    DOIs
    StatePublished - Jul 20 2009

    ASJC Scopus subject areas

    • Atomic and Molecular Physics, and Optics

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  • Cite this

    Cheong, F. C., Sun, B., Dreyfus, R., Amato-Grill, J., Xiao, K., Dixon, L., & Grier, D. G. (2009). Flow visualization and flow cytometry with holographic video microscopy. Optics Express, 17(15), 13071-13079. https://doi.org/10.1364/OE.17.013071