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Journal of Biomedical Optics • Open Access

Improved diffuse fluorescence flow cytometer prototype for high sensitivity detection of rare circulating cells in vivo
Author(s): Noah Pestana; Luke J. Mortensen; Judith M. Runnels; Dwayne Vickers; Shashi K. Murthy; Charles P. Lin; Mark J. Niedre

Paper 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.

Paper Details

Date Published: 5 July 2013
PDF: 8 pages
J. Biomed. Opt. 18(7) 077002 doi: 10.1117/1.JBO.18.7.077002
Published in: Journal of Biomedical Optics Volume 18, Issue 7
Show Author Affiliations
Noah Pestana, Northeastern Univ. (United States)
Luke J. Mortensen, Massachusetts General Hospital (United States)
Judith M. Runnels, Massachusetts General Hospital (United States)
Dwayne Vickers, Northeastern Univ. (United States)
Shashi K. Murthy, Northeastern Univ. (United States)
Charles P. Lin, Massachusetts General Hospital (United States)
Mark J. Niedre, Northeastern Univ. (United States)

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