title: Method for physiologic phenotype characterization at the single-cell level in non-interacting and interacting cells.
abstract: Intercellular heterogeneity is a key factor in a variety of core cellular processes including proliferation, stimulus response, carcinogenesis, and drug resistance. However, cell-to-cell variability studies at the single-cell level have been hampered by the lack of enabling experimental techniques. We present a measurement platform that features the capability to quantify oxygen consumption rates of individual, non-interacting and interacting cells under normoxic and hypoxic conditions. It is based on real-time concentration measurements of metabolites of interest by means of extracellular optical sensors in cell-isolating microwells of subnanoliter volume. We present the results of a series of measurements of oxygen consumption rates (OCRs) of individual non-interacting and interacting human epithelial cells. We measured the effects of cell-to-cell interactions by using the system's capability to isolate two and three cells in a single well. The major advantages of the approach are: 1. ratiometric, intensity-based characterization of the metabolic phenotype at the single-cell level, 2. minimal invasiveness due to the distant positioning of sensors, and 3. ability to study the effects of cell-cell interactions on cellular respiration rates.
authors.affiliation: Arizona State University, Biodesign Institute, Tempe, Arizona, USA. lkelbaus@asu.edu
authors.forename: Laimonas
authors.initials: L
authors.lastname: Kelbauskas
authors.forename: Shashanka P
authors.initials: SP
authors.lastname: Ashili
authors.forename: Jeff
authors.initials: J
authors.lastname: Houkal
authors.forename: Dean
authors.initials: D
authors.lastname: Smith
authors.forename: Aida
authors.initials: A
authors.lastname: Mohammadreza
authors.forename: Kristen B
authors.initials: KB
authors.lastname: Lee
authors.forename: Jessica
authors.initials: J
authors.lastname: Forrester
authors.forename: Ashok
authors.initials: A
authors.lastname: Kumar
authors.forename: Yasser H
authors.initials: YH
authors.lastname: Anis
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Paulson
authors.forename: Cody A
authors.initials: CA
authors.lastname: Youngbull
authors.forename: Yanqing
authors.initials: Y
authors.lastname: Tian
authors.forename: Mark R
authors.initials: MR
authors.lastname: Holl
authors.forename: Roger H
authors.initials: RH
authors.lastname: Johnson
authors.forename: Deirdre R
authors.initials: DR
authors.lastname: Meldrum
mesh_terms: D002450:Cell Communication / Q000502:physiology*; D018929:Cell Culture Techniques / Q000295:instrumentation; D002461:Cell Line, Transformed; D019069:Cell Respiration / Q000502:physiology; D006801:Humans; D016014:Linear Models; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D010101:Oxygen Consumption / Q000502:physiology*; D010641:Phenotype; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods*