CyTOF ™ Technology: Mass Spectrometry Solution to the Multi-parameter Flow Cytometry Challengeotential Role of Novel Imaging Modalities in Transplantation

Scott D. Tanner^1,2, Olga Ornatskyl, Vladimir Baranov², Dmitry Bandura².
¹University of Toronto, Toronto, ON, Canada; ²DVS Sciences Inc, ON, Canada

Learning Objectives:

1. Multi-parameter Flow Cytometry
2. Technology of Mass Cytometry
3. Immunological and Functional Assays

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 instnunent that "reads" the tag elements. Because there are many (100 or more) available stable isotopes, and the mass spectrometer provides exquisite resolution between detection channels, many parameters can be measured as easily as one. The mass cytometer itself is a specific configuration of an Inductively Coupled Plasma Time-of-Flight Mass Spectrometer, tailored for the analysis of up to 1000 cells per second for as many parameters as there are available stable isotope tags. The temporal evolution of the signal derived from DNA metalointercalation of each cell event informs on the integrity of the san1pled cell. The on-line processed data is compressed into *.fcs format that is compatible with third party cytometry software, so that the workflow "feels" like a flow cytometer, but reports on many parameters for single cells simultaneously without the need for spectral compensation. The ability to measure 30, 50 or more antigens simultaneously in single cells, without the need for compensation, will broaden the impact of flow cytometry, provide unprecedented precision and accuracy in the identitication of rare cell populations, and transform the proteomics and genomics research capability.

The technology has best application where many proteins or biomarkers arc to be determined simultaneously in single cells. For example, the simultaneous monitoring of multiple signaling pathways (e.g., phosphorylation events) under the influence of one or more agonists provides insight into the genesis of a diseased cell, or the pharmacokinetic characterization of a potential drug. We will show recent data, obtained in collaboration with the Nolan laboratory at Stanford University, on the simultaneous immunological and functional sUbpopu\ation of complex patient samples using a combination of 31 cell surface and intracellular antigens.