Bigfoot Multispectral High Speed Fluorescence Activated Cell Sorter

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We are requesting funds to assist us in purchasing a ThermoFisher Scientific Bigfoot spectral high speed cell sorter to expand and upgrade the equipment of the main Flow Cytometry Core Facility at the Albert Einstein College of Medicine. The Bigfoot is based on a remarkable new design that makes use of a principle known as full spectrum fluorescence analysis. This technology uses the full spectrum of emissions over a fixed detector array and an unmixing algorithm to greatly increase the number of fluorophores detected per cell, while eliminating most compensation issues that are major limitations of conventional flow cytometers. The optimized but flexible optical configuration makes the instrument robust and relatively easy to use, and permits detection of any fluorescence emission in the 400-900 nm range without requiring major adjustments of optical filters. The Bigfoot sorter’s optical design and unmixing algorithm enable the use of a wide array of dyes, including those with highly overlapping spectra—a feat not possible using conventional compensation-based cytometers and sorters. The problem of correct compensation of dyes with overlapping emission spectra, a major challenge for advanced multiparameter analysis, is mostly eliminated by the full spectrum approach of the Bigfoot. In addition to the unique spectral detection and processing, the Bigfoot provides extremely fast sort speeds for high yields and purity of specific cell populations. The instrument supports a range of sample outputs, including 6-way sorting into tubes and automated 8-way sorting into various format microtiter plates. A large cohort of NIH-funded investigators at Einstein working in diverse areas of biomedical science will benefit from access to this instrument, including the 24 major and 14 minor users specifically identified in this application. We plan to house and operate the Bigfoot sorter in our well established main Flow Cytometry Core Facility, where there will be ample expertise, oversight and attention to biosafety issues to ensure that the instrument is used correctly and to its full capacity. The Core Facility has an experienced senior faculty member (Dr. Steven Porcelli) as its supervisor, and employs two full time faculty level associates and two technicians to direct operations and provide technical supervision. There is also a range of internal consultants with expertise in areas of major relevance to the development of applications that will use the Bigfoot cell sorter. The Core Facility has a well-developed and carefully monitored financial plan, and receives support from the institution and from our NCI-funded Cancer Center to ensure its efficient and stable operation. By incorporating the Bigfoot spectral high speed cell sorter into this core laboratory, a large body of investigators pursuing diverse NIH-funded basic and translational research projects will have convenient and affordable access to this important new technology.