Automated platform for multiparameter stimulus response studies of metabolic activity at the single-cell level
2011
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.
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