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One Student's Introduction to the Applications, Principles, and Challenges of Mass Cytometry

By Pamella Tijerina Gann posted 03-10-2016 08:18

  

With the financial support of the Society of Toxicology (SOT) Supplemental Training for Education Program (STEP) Award administered by the Graduate Education Subcommittee, I had the opportunity to attend a training session by the University of Rochester Medical Center (URMC) Flow Core. Mass Cytometry is a relatively new technology that eliminates the weakness of emission signal overlap common in fluorescent, quantum dot, and other optical methods. Allowing the measurement of 30 or more antigens simultaneously in single cells, mass cytometry has provided unprecedented precision and accuracy in the identification of rare cell populations. 

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The next application deadline is May 1. Watch for the STEP currency that will be circulating at the SOT 2016 Annual Meeting.

The University of Rochester Medical Center obtained the DVS Sciences (acquired by Fluidigm in 2014) CyTOF Mass Cytometer in 2012 and has since become proficient at training and introducing users to the new technology. The flow core’s technical director, Matt Cochran, gives multiple lectures at several national conferences each year. His staff, especially Terry Wightman, was extremely helpful in explaining the novelties of CyTOF as well as the difficulties in implementation.

Mass cytometry, or CyTOF, is a variation of flow cytometry which uses stable isotopes as tags, rather than fluorochromes, in immunological assays with time-of-flight mass spectrometry detection. This enables an increase in the application of cytometry by allowing the combination of many more antibodies without emission signal overlap. Applicability involves experiments that have many proteins or biomarkers to be detected under multiple signaling pathways and few cells available for analysis. Using CyTOF, researchers have the unparalleled ability to phenotypically and functionally profile cells from normal and diseased states.  

Despite the unique capabilities of CyTOF, employing the technology isn’t without difficulties, which were discussed at great lengths during my training session. To become proficient in the technology, it is suggested one dedicated technician spend 6–8 months learning the system. Additionally, many researchers familiar with traditional flow cytometry are hesitant to accept the read-outs of CyTOF. This may be due to the lack of overlap with preceding techniques such as no equivalent forward or side scatter, slower acquisition rate (1,000 cells/second vs. 10,000 cells/second on conventional flow cytometers), and the destruction of cells during acquisition. Another inconvenience in the introduction of CyTOF is the initial investment in reagents as many laboratories currently have stocks of fluorescence-conjugated antibodies. This hurdle can be avoided by creating core reagent banks that enable scientists to share in the initial investment costs.

Advanced careers in core facilities offer an opportunity for scientists to engage in a vast variety of projects while maintaining their technical skills. My experience at URMC allowed me to peer into the world of a highly functional core facility and learn the basics behind the relatively novel technology of Mass Cytometry that isn’t offered at my institution’s core. Not only will my introduction to a new technique broaden my skills but also my awareness of the introduction of such technologies into large research facilities.

My department at the New York University Medical Center always looks for opportunities to stay relevant in the field of Toxicology. Since many of our studies involve multiple signaling pathways, many professors have requested an informational seminar on the application of CyTOF in our research. In the next year, I plan on holding such a seminar and educating my colleagues on the possible implementation of similar technologies in our department. In conclusion, my training session at URMC was an outstanding career development experience and I suggest other graduate students reach out to core facilities for such an opportunity.

  

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