Persistent Link:
http://hdl.handle.net/10150/193846
Title:
DEVELOPMENT AND ANALYSIS OF OPTICAL PH IMAGING TECHNIQUES
Author:
Lin, Yuxiang
Issue Date:
2010
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
The pH of tumors and surrounding tissues is a key biophysical property of the tumor microenvironment that affects how a tumor survives and how it invades the surrounding space of normal tissue. Research into tumorigenesis and tumor treatment is greatly dependent on accurate, precise, and reproducible measurements. Optical imaging is generally regarded as the best choice for non-invasive and high spatial resolution measurements. Ratiometric fluorescence imaging and fluorescence lifetime imaging microscopy (FLIM) are two primary ways for measuring tumor pH.pH measurements in a window chamber animal model using a ratiometric fluorescence imaging technique is demonstrated in this dissertation. The experimental setup, imaging protocols, and results are presented. A significantly varying bias was consistently observed in the measured pH. A comprehensive analysis on the possible error sources accounting for this bias is carried out. The result of analysis reveals that accuracy of ratiometric method is most likely limited by biological and physiological factors.FLIM is a promising alternative because the fluorescence lifetime is insensitive to the biological and physiological factors. Photon noise is the predominant error source of FLIM. The Fisher information matrix and the Cramér-Rao lower bound are used to calculate the lowest possible variance of estimated lifetime for time-domain (TD) FLIM. A statistical analysis of frequency-domain (FD) FLIM using homodyne lock-in detection is also performed and the probability density function of the estimated lifetime is derived. The results allow the derivation of the optimum experimental parameters, which yields the lowest variance of the estimated lifetime in a given period of imaging time. The analyses of both TD and FD-FLIM agree with results of corresponding Monte Carlo simulations.
Type:
text; Electronic Dissertation
Keywords:
accuracy and precision; error analysis; fluorescence lifetime; optical pH imaging; ratiometric imaging; statistical analysis
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Optical Sciences; Graduate College
Degree Grantor:
University of Arizona
Advisor:
Gmitro, Arthur F.
Committee Chair:
Gmitro, Arthur F.

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titleDEVELOPMENT AND ANALYSIS OF OPTICAL PH IMAGING TECHNIQUESen_US
dc.creatorLin, Yuxiangen_US
dc.contributor.authorLin, Yuxiangen_US
dc.date.issued2010en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractThe pH of tumors and surrounding tissues is a key biophysical property of the tumor microenvironment that affects how a tumor survives and how it invades the surrounding space of normal tissue. Research into tumorigenesis and tumor treatment is greatly dependent on accurate, precise, and reproducible measurements. Optical imaging is generally regarded as the best choice for non-invasive and high spatial resolution measurements. Ratiometric fluorescence imaging and fluorescence lifetime imaging microscopy (FLIM) are two primary ways for measuring tumor pH.pH measurements in a window chamber animal model using a ratiometric fluorescence imaging technique is demonstrated in this dissertation. The experimental setup, imaging protocols, and results are presented. A significantly varying bias was consistently observed in the measured pH. A comprehensive analysis on the possible error sources accounting for this bias is carried out. The result of analysis reveals that accuracy of ratiometric method is most likely limited by biological and physiological factors.FLIM is a promising alternative because the fluorescence lifetime is insensitive to the biological and physiological factors. Photon noise is the predominant error source of FLIM. The Fisher information matrix and the Cramér-Rao lower bound are used to calculate the lowest possible variance of estimated lifetime for time-domain (TD) FLIM. A statistical analysis of frequency-domain (FD) FLIM using homodyne lock-in detection is also performed and the probability density function of the estimated lifetime is derived. The results allow the derivation of the optimum experimental parameters, which yields the lowest variance of the estimated lifetime in a given period of imaging time. The analyses of both TD and FD-FLIM agree with results of corresponding Monte Carlo simulations.en_US
dc.typetexten_US
dc.typeElectronic Dissertationen_US
dc.subjectaccuracy and precisionen_US
dc.subjecterror analysisen_US
dc.subjectfluorescence lifetimeen_US
dc.subjectoptical pH imagingen_US
dc.subjectratiometric imagingen_US
dc.subjectstatistical analysisen_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineOptical Sciencesen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorGmitro, Arthur F.en_US
dc.contributor.chairGmitro, Arthur F.en_US
dc.contributor.committeememberUtzinger, Ursen_US
dc.contributor.committeememberClarkson, Eric W.en_US
dc.contributor.committeememberRouse, Andrew R.en_US
dc.identifier.proquest11164en_US
dc.identifier.oclc752261017en_US
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