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Massively multi-parameter single cell data by Mass Cytometry: the technology of its acquisition and networks for its interpretation

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Mass Cytometry brings the power, resolution, sensitivity and quantitative capabilities of atomic mass spectrometry to high throughput single cell analysis in order to address the challenges of multi-parameter, quantitative flow cytometry. Individual cells that have been immunologically stained with stable isotope tags are injected into the analytical instrument that ³reads² the tag elements. The cells are vaporized, atomized and ionized in a high temperature plasma, and the atomic composition of each cell including the metal tags is measured by time of flight mass spectrometry. Adapted from its long-time use in elemental analysis, the atomic mass spectrometer provides high sensitivity for many (up to 100) independent mass channels and offers the capability for absolute quantification. At present, 35 stable isotopes of the metals are available as tags, and we expect that another 30 will be available in the foreseeable future with the eventual potential for 100. The staining protocol is similar to that of flow cytometry, and the data output is in FCS format for porting into third party flow cytometry analysis software. Because the detection channels are independent, and the sensitivity to each probe is similar, the selection of staining panels is trivial. Accordingly, it is as easy to quantitatively analyze many parameters as a few, facilitated by the absence of need for compensation.

A high level introductory tutorial on the technology of element-labeling and analysis will be given. We will use data from our laboratory and that of our collaborators in the Nolan group at Stanford University, notably on determining differential immune and drug responses across a human hematopoietic continuum using 31 simultaneous cell surface and intracellular probes, to assess the current art in multidimensional data analysis.

Scott is co-founder and Chief Technology Officer (CTO) at DVS

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