In vivo multispectral, multiparameter, photoacoustic lymph flow cytometry with natural cell focusing, label-free detection and multicolor nanoparticle probes

Authors

  • Ekaterina I. Galanzha,

    1. University of Arkansas for Medical Sciences, Philips Classic Laser Laboratories, Winthrop P. Rockefeller Cancer Institute, Little Rock, Arkansas 72205
    2. Saratov State University, Institute of Optics and Biophotonics, Saratov 410012, Russia
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  • Evgeny V. Shashkov,

    1. University of Arkansas for Medical Sciences, Philips Classic Laser Laboratories, Winthrop P. Rockefeller Cancer Institute, Little Rock, Arkansas 72205
    2. Prokhorov General Physics Institute, Moscow 119991, Russia
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  • Valery V. Tuchin,

    1. Saratov State University, Institute of Optics and Biophotonics, Saratov 410012, Russia
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  • Vladimir P. Zharov

    Corresponding author
    1. University of Arkansas for Medical Sciences, Philips Classic Laser Laboratories, Winthrop P. Rockefeller Cancer Institute, Little Rock, Arkansas 72205
    • Philips Classic Laser Laboratories, University of Arkansas for Medical Sciences, 4301 West Markham St. #543, Little Rock, AR 72205-7199, USA
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Abstract

Compared with blood tests, cell assessment in lymphatics is not well-established. The goal of this work was to develop in vivo lymph tests using the principles of flow cytometry. Cells in living animals were counted by laser (420–2,300 nm) generation of photoacoustic (PA) signals in individual cells hydrodynamically focused by lymph valves into a single file flow, and using endogenous absorption as intrinsic cell-specific markers, or gold nanorods, nanoshells, and carbon nanotubes as multicolor probes. PA data were verified by high-speed transmission, photothermal, and fluorescent imaging. Counting of melanoma and immune-related cells in normal, apoptotic, and necrotic states in lymphatics in vivo was demonstrated to have the unprecedented sensitivity as one metastatic cell among millions of white blood cells. The time-resolved PA spectral identification of flowing cells was achieved using multicolor labels and laser pulses of different wavelengths and time delays. Multiparameter, noninvasive, portable flow cytometer can be used for preclinical studies on animals with the potential of translation to humans for in vivo PA mapping of colorless lymph vessels and sentinel nodes with simultaneous single cell detection and metastasis assessment without labeling or use of contrast dyes and/or novel low-toxic multicolor probes with different absorption spectra. © 2008 International Society for Advancement of Cytometry

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