CYTOF – HARNESSING MASS SPECTROMETRY FOR MULTIPARAMETRIC FLOW CYTOMETRY

Mass Cytometry (CyTOF) is a single-cell multiparametric antigen detection technology based on inductively coupled plasma mass spectrometry. It is an extension of Flow Cytometry in which antibodies are tagged with isotopically pure rare earth elements, rather than fluorophores, allowing simultaneous measurement of more than 40 parameters while circumventing the issue of spectral overlap. Hundreds of cells per second are passed through an argon plasma, ionizing the multi-atom metal tags, which are then analyzed by a time-of-flight mass spectrometer. Operation of the CyTOF is facilitated by the Flow Cytometry Unit at the Weizmann Institute of Science where researchers receive guidance regarding the technology, methodology, sample preparation, data acquisition and basic analysis. Conjugation service is provided by the Antibody Unit as well as management, maintenance and storage of a comprehensive institute-scale shared antibodies bank. Panel design and ordering of antibodies is accomplished by an automated algorithm, CyWIS, constructed by the Bioinformatics Unit and interlinked to the antibodies inventory. In-depth data analyses are also supported by the Bioinformatics Unit. Consequently, this backing allows easy assimilation of the platform to the researchers in a routine working manner, whilst maintaining good quality and high throughput reproducible data generation from the CyTOF. The lecture will illustrate this workflow and some of the CyTOF applications including profiling of blood, bone marrow, spleen, colon and ileum. Additionally, mass-tag cell barcoding will be described as well as the future technology of imaging mass cytometry.