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Principal Investigator: Iyadh Douagi
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2020
Award: $770,735
Funding agency: National Institute of Allergy and Infectious Diseases
In 2020 the project aims to use state of the art instrumentation to support the immediate research needs for COVID19 research in flow cytometry.
We are particularly interested in using the CytoFLEX LX Flow Cytometry Analyzer. It is a highly capable flow cytometry analyzer equipped with 6 lasers and 21 channels of fluorescent signal. For the first time ever, it will be housed in a custom BSC to add an additional level of biosafety protection. As demand for SARS-CoV-2 BSL3 operations increases, this instrument will be a great compliment to the FCS RTB platform, allowing for the safe and precise analysis of SARS-CoV-2 samples.
The instrument specifications:
6 laser capability including the following wavelengths:
- UV - 355nm
- Blue - 488 nm
- Yellow - Green 561nm
- Violet - 405nm
- Red - 638nm
- Infra-Red - 808nm
21 fluorescent detection channels
Internal 96-well plate loader for high throughput screening.
A compact size to fit into a BSC
An accompanying BSC that must be able to fit into the BSL3 lab that has a door width
of 38.5
A peristaltic pump sample delivery system to permit:
- calculation of absolute cell counts without the need for reference beads
- user defined fluidic rates, with increments of 1ul/min between 10 and 240 ml/min
High sensitivity and ability to resolve dim populations at 24 bits and 7 decades of
resolution and <30 MESF-FITC and <10 MESF-PE
Avalanche photodiodes (APDs) working from 200nm to 1100nm
Wavelength Division Multiplexing (WDM) detection module with solid-state, high
efficiency, low-noise detectors
A system that is serviceable on-site, without requiring the user to send the
instrument in for repair and periodic maintenance
In addition, we are interested in using a high-dimensional cell sorter. As an example, in order to identify and isolate new monoclonal antibodies that can be used as potential therapy for COVID19, we need to analyze rare lymphocyte subsets from convalescent COVID19 patients with frequencies (<0.05%). Without the added capacities in terms of lasers and detectors, it is not possible to efficiently detect nor isolate those cells. When optimized, this strategy can easily be shared within a broad portfolio of DIR projects aiming to develop therapeutic strategies for COVID19.
We also aim to upgrade the BD FACSAria Fusion SO Laser Option. As we are currently, limited to 4 lasers and 11 colors, this upgrade will permit us to extend the capacity of the current BD FACSAria Fusion platform to 5 lasers and 18 colors, to make it closely match the other instruments. Many DIR investigators currently using the RTB will be able to take advantage of this added capacity for cell sorting within the BSL3 lab.
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