Improved diffuse fluorescence flow cytometer prototype for high sensitivity detection of rare circulating cells in vivo

Noah Pestana; Luke J. Mortensen; Judith M. Runnels; Dwayne Vickers; Shashi K. Murthy; Charles P. Lin; Mark J. Niedre

doi:10.1117/1.JBO.18.7.077002

5 July 2013 Improved diffuse fluorescence flow cytometer prototype for high sensitivity detection of rare circulating cells in vivo

Noah Pestana, Luke J. Mortensen, Judith M. Runnels, Dwayne Vickers, Shashi K. Murthy, Charles P. Lin, Mark J. Niedre

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Journal of Biomedical Optics, Vol. 18, Issue 7, 077002 (July 2013). https://doi.org/10.1117/1.JBO.18.7.077002

Abstract

Detection and enumeration of rare circulating cells in mice are important problems in many areas of preclinical biomedical research. Recently, we developed a new method termed “diffuse fluorescence flow cytometry” (DFFC) that uses diffuse photons to increase the blood sampling volume and sensitivity versus existing in vivo flow cytometry methods. In this work, we describe a new DFFC prototype with approximately an order-of-magnitude improvement in sensitivity compared to our previous work. This sensitivity improvement is enabled by a number of technical innovations, which include a method for the removal of motion artifacts (allowing interrogation of mouse hindlegs that was less optically attenuating versus the tail) and improved collection optics and signal preamplification. We validated our system first in limb mimicking optical flow phantoms with fluorescent microspheres and then in nude mice with fluorescently labeled mesenchymal stem cells at injected concentrations of 5×10 ³ cells/mL . In combination, these improvements resulted in an overall cell counting sensitivity of about 1 cell/mL or better in vivo.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Noah Pestana, Luke J. Mortensen, Judith M. Runnels, Dwayne Vickers, Shashi K. Murthy, Charles P. Lin, and Mark J. Niedre "Improved diffuse fluorescence flow cytometer prototype for high sensitivity detection of rare circulating cells in vivo," Journal of Biomedical Optics 18(7), 077002 (5 July 2013). https://doi.org/10.1117/1.JBO.18.7.077002

Published: 5 July 2013

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