Cell sorting flow cytometer with spectral detection

About This Grant

Project Summary This shared instrument grant requests funds for an Aurora CS for fluorescence-based cell sorting, to be installed at Yale University. Flow Cytometry is a critical resource for biomedical research. Nearly every discipline in the biologic sciences uses this technology that is critical for a variety of applications downstream of the sorting of cells. There is a growing demand for high-parameter flow cytometry at Yale to characterize biological samples as completely as possible. As the amount of published phenotypic information on distinct cell types and their subsets continues to increase, the number of parameters that is then needed to uniquely identify and distinguish diverse cell subset is likewise growing at a rapid pace. However, the number of cells within a sample can sometimes be small and as a result, an increasing number of studies require higher parameter panels during cell sorting to collect deep phenotypic information with small samples that cant also be analyzed separately at another instrument. For these reasons the Aurora CS is an ideal instrument to meet these needs because it 1) has full spectral detection with more than 60 sensitive detectors that generate spectral signatures that can be computationally “unmixed”, 2) enables measurement of over 30 cellular parameters simultaneously, 3) greatly improves the ability to distinguish multiple fluorescent proteins such as GFP, YFP or RFPs with overlapping emissions, or any other challenging dye combination, 4) identifies spectral signatures of unique cell-type specific autofluorescence that can then be computationally removed, solving a perennial problem with highly autofluorescent samples such as those derived from kidneys, lungs or intestines, and 5) further offers the ability to sort objects as small as 100 nm, such as EVs, synaptosomes or platelets. The requested equipment would replace a heavily used but aging BD FACSAria. The vendor will soon cease to offer service contracts on this model. Since this FACSAria is the only cell-sorting flow cytometer available in its building, any equipment breakdown could result in delays that would significantly impede ongoing research. It is critical that the existing instrument be replaced with a reliable cell-sorter that can be repaired in a timely fashion, preferably one that can be operated independently by investigators when staff is not available. The Aurora CS will serve as an ideal “future proof” cell sorter in a demanding location while addressing multiple other unmet needs on campus. Thus, the requested equipment will empower Yale researchers to interrogate complex biology with more challenging phenotypic panels, accelerating the pace of research while engendering greater insights and deeper knowledge.