MRI: Acquisition of FACSARIA-II, a next-generation high-speed cell sorter for a Flow Cytometry Core supporting interdisciplinary research and training

University Of Massachusetts Amherst, Amherst MA

Investigators

Abstract

Flow cytometry is a powerful analytic technique that facilitates the characterization, quantification and/or isolation of cell populations based on innate or manipulated cellular attributes. Labeling cells externally or internally with fluorescently-conjugated antibodies or fluorescent vital dyes, or by genetically expressing fluorescent reporter molecules, all represent strategies for defining cell populations that may be differentiated using a flow cytometer.In recent years, BectonDickinson has made significant technological improvements, leading the field in creating the next-generation of cell sorters with its FACSAria (4) platform. A newer, user-friendly, bench-top high-speed cell sorter, the FACSAriaII, has digital electronics that allows high-speed analyses (70,000 events/second) based on up to 14 parameters, including cell size and internal complexity, with concomitant high-speed, simultaneous sorting of two or four populations, saving time and resources for users. Sample nozzles are easily interchanged to accommodate sorting large cells. Its fixed-laser alignment increases sensitivity, minimizes start-up time, improves reproducibility between experiments and enables automated daily quality control. It is capable of sorting cells aseptically for subsequent cell culture. The FACSAriaII platform is upgradeable, permitting economic configuration at the outset and retaining the flexibility to meet evolving user needs in the future. High-speed sorting, large cell size accommodation, increased sensitivity and upgradability make the FACSAriaII the best possible choice now and an excellent long-term investment. This proposal describes the research of nine PIs as major users of a FACSAriaII, and an additional 10 minor users. Their projects, are broadly grouped by their requirements for one or more of the unique capabilities intrinsic to the FACSAriaII: i) high-speed sorting of unique target cell populations; ii) simultaneous, high-speed enrichment of multiple cell populations for further in vivo or ex vivo utilization; iii) high-speed sorting requiring multi-parameter stratification. Many of these PIs have optimized protocols in place, ensuring data will be obtained almost immediately upon acquiring a FACSAriaII. Reagent development is another important goal of many of the PIs, and acquiring a FACSAriaII will provide them with the capacity to advance and accelerate discovery for entire fields of research. Chemical engineers will modify and analyze plant cells, on a per cell basis, to improve their production of cancer-treating compounds, or engineer bacterial quorum sensing for synthetic biology and industrial biotechnology. Biologists will be able to better understand the genes that regulate the structural dynamics of plant growth and chemists will use bacterial expression systems to generate and screen for synthetic allosteric ?switches? that can regulate cell death. Polymer scientists will track the efficiency and consequences of synthetic payload uptake into viable cells, while immunologists will recover and manipulate rare cell populations, transferring them from one host to another to better define mechanisms of disease, identify novel therapeutic targets, or advance vaccine development. Ongoing, diverse and, in many cases, interdisciplinary research will be strengthened and accelerated by the capabilities of a FACSAriaII high-speed cell sorter. Many of the proposed research projects have applied objectives including potential translational benefits, such as identifying novel therapeutic targets and providing proof-of-principle data for novel delivery of payload. Additionally, some of these projects may have industrial or commercial applications. On-site access to a FACSAriaII will hasten progress toward these objectives. The group of major and minor users collectively trains many undergraduate and graduate students, and many of these through grant-funded interdisciplinary education and training, such as the University of Massachusetts/ Amherst Institute for Cellular Engineering and the Chemistry/Biology Interface. Looking forward, through outreach to sister colleges in the Five College Consortium, the Flow Core Facility will provide expanded opportunities for training undergraduate students in flow cytometric techniques, including students from Mt. Holyoke and Smith College, two all-women colleges that prepare a significant percentage of their undergraduate enrollment for advanced training in the STEM fields. Specifically, in conjunction with Smith College's Summer Science and Engineering Program, offered each year to high school-aged young women from across the country, a flow cytometry and cell sorting research module will be developed to introduce the next generation of women scientists to the power of single-cell analysis. Looking back, the University of Massachusetts/Amherst has a long-standing reputation for recruiting and retaining under-represented groups, in part through its lead role in the Northeast Alliance for Graduate Education and the Professoriate. This five-institution organization is committed to expanding opportunities for scientific graduate education and career advancement for under-represented groups. In 2008, the Cargnegie Foundation recognized the outreach efforts of the University of Massachusetts/Amherst when it designated UMass a Community Engaged University. The FACSAriaII, an instrument that combines powerful single-cell sorting and analysis with an accessible, user-friendly design will play an invaluable role in continuing these efforts.