Apparatus for the isolation of specific primary cells from tissue

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NSERC-RTI funds are requested to acquire a platform for conducting fluorescence activated cell sorting (FACS) to isolate primary cells from tissue. Our request is for a state-of-the-art FACS from BD Biosciences (BD FACS MelodyT 2 Laser), which will be a unique addition to be housed in the Farquharson Life Science Building at York University. FACS is the gold standard method to isolate primary cells, based on its precision, flexibility and reliability. It is ideal for the isolation of adult stem cells and fluorescently marked cells from the complex multi-cell population released by tissue digestion. This sophisticated equipment will enable the discrimination between distinct live cell populations based on discrete fluorescence labelling, and then will sort and collect these populations to grow in culture or to be used for further assessments. Importantly, the requested FACS is ideal for use laboratory use: it is user-friendly and does not require a dedicated technician. The use of FACS is integral to the NSERC-funded research programmes of primary applicant Dr. Anthony Scimè and co-applicants Drs. Tara Haas and Emilie Roudier, who are in the School of Kinesiology and Health Science and members of the York University Muscle Health Research Centre (MHRC). The applicants are diverse with respect to gender, ethnicity and career stage, thus providing distinct perspectives and role models to trainees. Each applicant's current and future research hinges on the effective purification of specific primary cell populations from the mixed cell types present within tissues. Currently, there is a dire need for this apparatus, as York University does not possess a 2-laser FACS. At present, FACS must be performed at facilities located several kilometers distant. The lag-time spent transporting the materials (pre and post sort) causes the loss of integrity and excess death of isolated cells. Those facilities also are heavily used and have limited availability. Crucially, these are issues that significantly impede the research progress of the applicants and HQP. The FACS that we intend to purchase will not only be available to meet the immediate and long-term research needs the applicants, but those of several research groups at York University including the Muscle Health research community and others in the Faculties of Health, Science and Engineering. Importantly, many HQP will benefit from the use of this equipment. Trainees will become proficient using the gold standard foundational cell biological approach of FACS to investigate specific cell populations. Having this user-friendly FACS at York will enable more trainees to incorporate this sophisticated technique into their projects, which will elevate the quality and potential impact of their data. It will attract new trainees to join our research programs and will facilitate collaborations of HQP with other researchers.