Welsh, Joshua, Horak, Peter, Wilkinson, James, Ford, Verity J., Jones, Jennifer C., Smith, David, Holloway, Judith and Englyst, Nicola (2019) FCMPASS software aids extracellular vesicle light scatter standardization’. Cytometry Part A. (doi:10.1002/cyto.a.23782).
Abstract
The study of extracellular vesicles (EVs) is a rapidly growing field due to their great potential in many areas of clinical medicine including diagnostics, prognostics, theranostics, and therapeutics. Flow cytometry is currently one of the most popular methods of analysing EVs due to it being a high-throughput, multi-parametric technique, that is readily available in the majority of research labs. Despite its wide use, few commercial flow cytometers are designed specifically for the detection of EVs. Many flow cytometers used for EV analysis are working at their detection limits and are unable to detect the majority of EVs. Currently, very little standardisation exists for EV flow cytometry which is an issue because flow cytometers vary considerably in the way they collect scattered or fluorescent light from particles being interrogated. This makes published research hard-to-interpret, compare,and in some cases, impossible to reproduce. Here we demonstrate a method of flow cytometer light scatter standardisation, utilising flow cytometer post-acquisition analysis software (FCMPASS). FCMPASS is built upon Mie theory and enables the approximation of flow cytometer geometric parameters either by analyzing beads of known diameter and refractive index or by inputting the collection angle if known. The software is then able to create a scatter-diameter curve and scatter-refractive index curve that enables researchers to convert scattering data and instrument sensitivity into standardised units. Furthermore, with the correct controls, light scatter data can be converted to diameter distributions or refractive index distributions. FCMPASS therefore offers a freely available and ergonomic method of standardizing and further extending EV characterization using flow cytometry.
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- Current Faculties > Faculty of Engineering and Physical Sciences > Zepler Institute for Photonics and Nanoelectronics > Fibre and Systems Group
Zepler Institute for Photonics and Nanoelectronics > Fibre and Systems Group - Current Faculties > Faculty of Engineering and Physical Sciences > Zepler Institute for Photonics and Nanoelectronics > Nanophotonics Group > Photonic Systems Circuits and Sensors Group
Zepler Institute for Photonics and Nanoelectronics > Nanophotonics Group > Photonic Systems Circuits and Sensors Group - Faculties (pre 2018 reorg) > Faculty of Natural and Environmental Sciences (pre 2018 reorg) > Institute for Life Sciences (pre 2018 reorg)
Current Faculties > Faculty of Environmental and Life Sciences > Institute for Life Sciences > Institute for Life Sciences (pre 2018 reorg)
Institute for Life Sciences > Institute for Life Sciences (pre 2018 reorg)
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