Abstract

Applications for Small Particle Analysis Using High Sensitivity Flow and Imaging Flow Cytometry

Haley Pugsley, Manager and Senior Scientist, Luminex Corporation

Research in the fields of virology, microbiology, nanoparticles, and extracellular vesicles (EVs) has shown tremendous growth over the past few years. Viruses, which range in size from 17 nm to ~1.5 microns, small bacteria, nanoparticles, and small EVs such as exosomes (less than 150 nm in diameter), are all on the subcellular scale. Quantifying and characterizing small particles in a reproducible and reliable manner is challenging due to their small size. In this presentation, we will demonstrate the capabilities for small particle applications from Amnis® CellStream^® and Amnis^® ImageStream^®X Mk II. The Amnis CellStream system has the advantage of high throughput flow cytometry with higher sensitivity to small particles due to the CCD-based, time-delay-integration image capturing system. Here, we will present immunophenotyping data from EVs. The Amnis ImageStreamX Mk II that combines the quantitative power of flow cytometry with microscopy in one system has a High Gain mode to increase the sensitivity for small particles by adjusting the CCD-camera to a higher gain setting, increasing the signal obtained from the small particles while minimizing the noise. In this portion of the presentation, examples of High Gain mode using murine leukemia virus-sfGFP will be shown.