Apr 12, 2012
from 09:00 AM to 11:00 AM
|Where||LCCC, Pagano Conference Room|
|Contact Name||Lori Smith|
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Mass Cytometry (also referred to as CyTOF, cytometry by time-of-flight) applies the analytical characteristics of atomic mass spectrometry to the challenges of multi-parameter flow cytometry. With a 30-year history as the state of the art in the quantitative determination of the elemental composition of matter, the significant attributes that translate to cell analysis are the linear dynamic range, up to 100 independent (mass) detection channels and sensitivity that is linearly dependent on the number of metal atoms appended to the protein or biomarker.
This talk will present the fundamentals of the technology, and highlight those design decisions that allow it to emulate fluorescent flow cytometers. Briefly, probes (antibodies, DNA-intercalators) are labeled uniquely with enriched stable isotopes of the transition elements. Cell samples are typically fixed, permeabilized and stained in a familiar manner, and are injected individually into a high temperature (7000 K) plasma where they are vaporized, atomized and ionized. The atomic ions, including the tagging isotopes, are mass analyzed and counted. Single cell events are recognized in real-time based on the transient profile of the signals, and the data is converted to the *.fcs format that is compatible with third party flow cytometry software. Up to 1000 cells per second can be analyzed for 40+ proteins simultaneously, without the need for spectral overlap compensation. Various approaches to the analysis of such massively multi-variate data will be presented in the context of current applications of the technology. Our own work on neural network identification of rare cell populations is complemented by the ground-breaking work on the characterization of continua in the hematopoietic system (Bendall, Simonds et al, Science, May 2011) and in CD8+ T-cell populations (Newell et al, Immunity, January 2012), and massive multiplexing through isotopic bar-coding as demonstrated by Bodenmiller and Zunder (paper pending).