The Nano Flow Cytometry Pipeline at the Cytometry Platfrom, CBUTechS
Cytometry POSTER – Nano flow cytometry pipeline
In order to bring more insight into the mechanisms of transfer of biological molecules such as proteins and RNA between cells, and to study signaling pathways, it is necessary to detect, characterize and quantify extracellular vesicles (EVs) and virions. However, this is challenging due to the small size of these particles (e.g. EVs range of diameter from 40 to 130 nm) and to the lack of specific staining reagents for purification of viruses. The resolution of conventional cytometers and sorters enables distinguishing or sorting particles with a diameter of minimum 300nm.
The Cytometry platform of the CB UTechS has collaborated with scientific groups of Virology and Microbiology departments to develop a pipeline that pushes the limits of conventional approaches to allow analysis and sorting of subcellular particles.
With the “Nano Flow Cytometry pipeline” that we developed, it is possible to visualize nanoparticles in the 40–850 nm size range. This pipeline takes advantage of a combination of an adapted optical bench and settings of a jet-in-air cell sorter and the new nano analyzer system. The NanoAnalyzerTM (NanoFCM) technology allows detecting and quantifying EVs and viruses in a fast and reliable way without the need of any fluorescent read out. In parallel, with the optimized high-speed sorting (Moflo AstriosTM, Beckman Coulter) that combines sample labelling, specific optical parameters, and finely defined laser power settings, we have demonstrated our capacity to detect and sort retroviral particles.