Advisor(s)
Mark J. Niedre
Contributor(s)
Shashi Murthy, Charles A. DiMarzio
Date of Award
2011
Date Accepted
8-2011
Degree Grantor
Northeastern University
Degree Level
M.S.
Degree Name
Master of Science
Department or Academic Unit
College of Engineering, Department of Electrical and Computer Engineering
Keywords
electrical engineering, biomedical engineering, optics, in vivo flow cytometry, optical and photonic sensors and systems, single cell analysis
Disciplines
Electrical and Computer Engineering | Engineering
Abstract
Accurate quantification of circulating cell populations is important in many areas of preclinical and clinical biomedical research including the study of metastasized cancers, T-Lymphotocyes and hematopoietic stem cells. Normally this is done either by extraction and analysis of small blood samples or more recently using microscopy-based in vivo fluorescence flow cytometry. In this thesis, a new technological approach to this problem is described using detection of diffuse fluorescent light from relatively large blood vessels in vivo. The `tomographic diffuse fluorescence flow cytometer' (TDFFC) uses modulated lasers to illuminate a mouse limb and an array of optical fibers coupled to a high-sensitivity photomultiplier tube array operating in photon counting mode to detect weak fluorescence signals from cells. It is first demonstrated that the TDFFC instrument is capable of detecting fluorescent microspheres and Vybrant-DiD labeled cells with excellent accuracy in an optical flow phantom with similar size, optical properties, linear flow rates and autofluorescence as a mouse limb. Preliminary data demonstrating that the TDFFC is capable of detecting circulating cells in nude mice in vivo is also shown. Finally, a number of methods for performing coarse tomographic localization of fluorescent cells within the cross-section of a mouse limb using TDFFC data sets are described, and the feasibility of this approach is demonstrated using in vitro data sets. In principle, this device would allow interrogation of the whole blood volume of a mouse in minutes, with several orders of magnitude sensitivity improvement compared with current approaches.
Document Type
Master's Thesis
Rights Information
copyright 2011
Rights Holder
Eric William Zettergren
Permanent URL
Recommended Citation
Zettergren, Eric William, "Tomographic diffuse fluorescence flow cytometry for enumeration of rare circulating cells in vitro and in vivo" (2011). Electrical and Computer Engineering Master's Theses. Paper 69. http://hdl.handle.net/2047/d20002081
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