COVID-19 Instrumentation in the RTB

National Institute Of Allergy And Infectious Diseases

Investigators

Linked publications, trials & patents

Abstract

The project aims to use state of the art instrumentation to support the immediate research needs for COVID19 research in flow cytometry. We are interested in developing new capacities for high-dimensional cell sorting requiring the precise identification of rare cell subsets based on multiple parameters and lasers. Current cell sorters are not matched to the evolving analytical demands in the field of high-parameter flow cytometry. Increasing number of RTB users are transitioning to complex assays for cell analysis. However, our sorters currently lack the ability to translate these panels directly into cell sorting for further downstream assays. The acquisition of the High-end cell sorter will contribute to open new possibilities in cell sorting based on up to 30 parameter and will allow for the first time simultaneous six-way sorting. These combined features are key for the successful isolation of rare-cells within often unique clinical samples. 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. In addition, recognizing the risk of aerosol exposure during cell sorting, biosafety guidelines were developed during the last two decades. The use of aerosol management systems (AMS) and enclosure of cell sorters within a biosafety cabinet (BSC) are today well-established procedures for infectious cell sorting. In contrast, analyzers are perceived to be at lower risk for aerosol generation. We recently demonstrated that aerosols can be generated during certain operations and proposed BSC enclosure for analysis of infectious samples (Aspland et al 2021). We aim to setup and a novel cytometry platform contained in a Class II BSC for safe analysis of infectious samples in BSL-3 laboratory. The integration of this platform into a BSL3 lab environment will allow DIR investigators to perform analysis of cellular perturbations in live cells exposed to SARSCov2. These findings are expected to provide new insights into cellular mechanisms of host pathogen interactions and to accelerate development of novel therapeutics for COVID19. Finally, recent advances in spectral technology allows to collect light through the entire spectrum and to objectively manage overlap between fluorescence spectra thereby improving data accuracy compared to conventional flow cytometry. The acquisition of a spectral cell sorter will (1) allow matching and direct transition of complex experimental protocols (based on 40 colors and beyond) currently used by multiple DIR investigators using the Cytek Spectral Analyzer platform. (2) By providing unique cell sorting capabilities based on ultra-deep immunophenotyping, will allow to increase understanding of immunes responses to pathogens including SARS-CoV-2, while ensuring continuity in developing this cutting-edge technology.