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Cytometry of Reaction Rate Constant (Crrc): Optimization of the Technique and Development of a Novel Protocol to Assess Cell Population Heterogeneity Based on Aldh1a1 Activity in Vitro

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dc.contributor.advisorKrylov, Sergey N.
dc.contributor.authorNebbioso, Giammarco
dc.date.accessioned2023-12-08T14:35:14Z
dc.date.available2023-12-08T14:35:14Z
dc.date.issued2023-12-08
dc.date.updated2023-12-08T14:35:14Z
dc.degree.disciplineBiology
dc.degree.levelMaster's
dc.degree.nameMSc - Master of Science
dc.description.abstractCytometry of Reaction Rate Constant (CRRC) is a novel analytical technique which aims to study cellular heterogeneity based on the activity of enzymatic reactions. In the past, CRRC was able to assess cellular heterogeneity in vitro by investigating the activity of ABC-transporter enzymes. However, CRRC showed poor robustness to highly motile cells. Here, I report on the development of a workflow to make CRRC robust to cell motility. The novel workflow was used to develop a protocol for in vitro CRRC studies of aldehyde dehydrogenase 1A1 (ALDH1A1)-based cell heterogeneity. The data collected suggested a potential positive correlation between the activity of ALDH1A1 and the age of a cell line. Finally, I demonstrated the robustness of the new CRRC ALDH1A1 assay to a 20% change in the initial substrate concentration. Overall, these studies confirm the potential for CRRC to become a reliable analytical tool for studies of reaction-based cell heterogeneity.
dc.identifier.urihttps://hdl.handle.net/10315/41687
dc.languageen
dc.rightsAuthor owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.
dc.subjectBiology
dc.subjectAnalytical chemistry
dc.subjectMolecular biology
dc.subject.keywordsCytometry
dc.subject.keywordsFluorescence microscopy
dc.subject.keywordsClassical microscopy
dc.subject.keywordsReaction rate constant
dc.subject.keywordsCell heterogeneity
dc.subject.keywordsTime-lapse experiment
dc.subject.keywordsMichaelis-Menten equation
dc.subject.keywordsFirst order kinetics
dc.subject.keywordsKinetics
dc.titleCytometry of Reaction Rate Constant (Crrc): Optimization of the Technique and Development of a Novel Protocol to Assess Cell Population Heterogeneity Based on Aldh1a1 Activity in Vitro
dc.typeElectronic Thesis or Dissertation

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