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Proceedings Paper

Automated platform for multiparameter stimulus response studies of metabolic activity at the single-cell level
Author(s): Shashanka P. Ashili; Laimonas Kelbauskas; Jeff Houkal; Dean Smith; Yanqing Tian; Cody Youngbull; Haixin Zhu; Yasser H. Anis; Michael Hupp; Kristen B. Lee; Ashok V Kumar; Juan Vela; Andrew Shabilla; Roger H. Johnson; Mark R. Holl; Deirdre R. Meldrum
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Paper Abstract

We have developed a fully automated platform for multiparameter characterization of physiological response of individual and small numbers of interacting cells. The platform allows for minimally invasive monitoring of cell phenotypes while administering a variety of physiological insults and stimuli by means of precisely controlled microfluidic subsystems. It features the capability to integrate a variety of sensitive intra- and extra-cellular fluorescent probes for monitoring minute intra- and extra-cellular physiological changes. The platform allows for performance of other, post- measurement analyses of individual cells such as transcriptomics. Our method is based on the measurement of extracellular metabolite concentrations in hermetically sealed ~200-pL microchambers, each containing a single cell or a small number of cells. The major components of the system are a) a confocal laser scan head to excite and detect with single photon sensitivity the emitted photons from sensors; b) a microfluidic cassette to confine and incubate individual cells, providing for dynamic application of external stimuli, and c) an integration module consisting of software and hardware for automated cassette manipulation, environmental control and data collection. The custom-built confocal scan head allows for fluorescence intensity detection with high sensitivity and spatial confinement of the excitation light to individual pixels of the sensor area, thus minimizing any phototoxic effects. The platform is designed to permit incorporation of multiple optical sensors for simultaneous detection of various metabolites of interest. The modular detector structure allows for several imaging modalities, including high resolution intracellular probe imaging and extracellular sensor readout. The integrated system allows for simulation of physiologically relevant microenvironmental stimuli and simultaneous measurement of the elicited phenotypes. We present details of system design, system characterization and metabolic response analysis of individual eukaryotic cells.

Paper Details

Date Published: 14 February 2011
PDF: 12 pages
Proc. SPIE 7929, Microfluidics, BioMEMS, and Medical Microsystems IX, 79290S (14 February 2011); doi: 10.1117/12.875438
Show Author Affiliations
Shashanka P. Ashili, Arizona State Univ. (United States)
Laimonas Kelbauskas, Arizona State Univ. (United States)
Jeff Houkal, Arizona State Univ. (United States)
Dean Smith, Arizona State Univ. (United States)
Yanqing Tian, Arizona State Univ. (United States)
Cody Youngbull, Arizona State Univ. (United States)
Haixin Zhu, Arizona State Univ. (United States)
Yasser H. Anis, Cairo Univ. (Egypt)
Michael Hupp, Arizona State Univ. (United States)
Kristen B. Lee, Arizona State Univ. (United States)
Ashok V Kumar, Arizona State Univ. (United States)
Juan Vela, Arizona State Univ. (United States)
Andrew Shabilla, Arizona State Univ. (United States)
Roger H. Johnson, Arizona State Univ. (United States)
Mark R. Holl, Arizona State Univ. (United States)
Deirdre R. Meldrum, Arizona State Univ. (United States)

Published in SPIE Proceedings Vol. 7929:
Microfluidics, BioMEMS, and Medical Microsystems IX
Holger Becker; Bonnie L. Gray, Editor(s)

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