11 February 2011 A novel method for multiparameter physiological phenotype characterization at the single-cell level
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Non-genetic intercellular heterogeneity has been increasingly recognized as one of the key factors in a variety of core cellular processes including proliferation, stimulus response, carcinogenesis and drug resistance. Many diseases, including cancer, originate in a single or a few cells. Early detection and characterization of these abnormal cells can provide new insights into the pathogenesis and serve as a tool for better disease diagnosis and treatment. We report on a novel technology for multiparameter physiological phenotype characterization at the single-cell level. It is based on real-time measurements of concentrations of several metabolites by means of extracellular optical sensors in microchambers of sub-nL volume containing single cells. In its current configuration, the measurement platform features the capability to detect oxygen consumption rate and pH changes under normoxic and hypoxic conditions at the single-cell level. We have conceived, designed and developed a semi-automated method for single-cell manipulation and loading into microwells utilizing custom, high-precision fluid handling at the nanoliter scale. We present the results of a series of measurements of oxygen consumption rates (OCRs) of single human metaplastic esophageal epithelial cells. In addition, to assess the effects of cell-to-cell interactions, we have measured OCRs of two and three cells placed in a single well. The major advantages of the approach are a) multiplexed characterization of cell phenotype at the single-cell level, b) minimal invasiveness due to the distant positioning of sensors, and c) flexibility in terms of accommodating measurements of other metabolites or biomolecules of interest.
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Laimonas Kelbauskas, Laimonas Kelbauskas, Shashanka Ashili, Shashanka Ashili, Jeff Houkal, Jeff Houkal, Dean Smith, Dean Smith, Aida Mohammadreza, Aida Mohammadreza, Kristen Lee, Kristen Lee, Ashok Kumar, Ashok Kumar, Yasser Anis, Yasser Anis, Tom Paulson, Tom Paulson, Cody Youngbull, Cody Youngbull, Yanqing Tian, Yanqing Tian, Roger Johnson, Roger Johnson, Mark Holl, Mark Holl, Deirdre Meldrum, Deirdre Meldrum, } "A novel method for multiparameter physiological phenotype characterization at the single-cell level", Proc. SPIE 7902, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX, 79021Q (11 February 2011); doi: 10.1117/12.875483; https://doi.org/10.1117/12.875483

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