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Method for physiologic phenotype characterization at the single-cell level in non-interacting and interacting cells. J Biomed Opt 2012 Mar;17(3):037008

Date

04/17/2012

Pubmed ID

22502580

Pubmed Central ID

PMC3602818

DOI

10.1117/1.JBO.17.3.037008

Scopus ID

2-s2.0-84864922062   17 Citations

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.

Author List

Kelbauskas L, Ashili SP, Houkal J, Smith D, Mohammadreza A, Lee KB, Forrester J, Kumar A, Anis YH, Paulson TG, Youngbull CA, Tian Y, Holl MR, Johnson RH, Meldrum DR

Author

Roger H. Johnson PhD Associate Professor in the Biophysics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Cell Communication
Cell Culture Techniques
Cell Line, Transformed
Cell Respiration
Humans
Linear Models
Microfluidic Analytical Techniques
Microscopy
Oxygen Consumption
Phenotype
Single-Cell Analysis