Automated platform for multiparameter stimulus response studies of metabolic activity at the single-cell level

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 Meldrum

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMicrofluidics, BioMEMS, and Medical Microsystems IX
DOIs
StatePublished - 2011
EventMicrofluidics, BioMEMS, and Medical Microsystems IX - San Francisco, CA, United States
Duration: Jan 23 2011Jan 25 2011

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume7929
ISSN (Print)0277-786X

Other

OtherMicrofluidics, BioMEMS, and Medical Microsystems IX
CountryUnited States
CitySan Francisco, CA
Period1/23/111/25/11

Keywords

  • Single cell analysis
  • and transmembrane flux measurements
  • automated single cell systems
  • cell-cell interaction
  • eukaryotic cells
  • hypoxia
  • microfluidics
  • microsensors

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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  • Cite this

    Ashili, S. P., Kelbauskas, L., Houkal, J., Smith, D., Tian, Y., Youngbull, C., Zhu, H., Anis, Y. H., Hupp, M., Lee, K. B., Kumar, A. V., Vela, J., Shabilla, A., Johnson, R. H., Holl, M. R., & Meldrum, D. (2011). Automated platform for multiparameter stimulus response studies of metabolic activity at the single-cell level. In Microfluidics, BioMEMS, and Medical Microsystems IX [79290S] (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 7929). https://doi.org/10.1117/12.875438