Abstract

Expanding Capabilities for Small Particle Analysis Using Imaging Flow Cytometry and Full Spectral Flow Cytometry

Stephanie Brunelle, Senior Product Manager, Cytek BioSciences

In this presentation, we will discuss how to expand analysis options to study small particles and extracellular vesicles (EVs) using Cytek’s solutions, including the Cytek® Amnis® ImageStream®, Cytek® Aurora™, and Cytek® Northern Lights™ systems.  Research in microbiology, virology, nanoparticles, and EVs has grown tremendously over the past few years. Small bacteria, viral particles, nanoparticles, and small EVs, such as exosomes (less than 150 nm in diameter), are all on the subcellular scale. Detection, quantification, and characterization of small objects in a reproducible and reliable manner is challenging due to their small size and interference with debris. We will demonstrate the capabilities of the Cytek® systems for small particle applications. The Cytek® Amnis® ImageStream®X Mk II system, which combines the quantitative power of flow cytometry with microscopy in one system, has a “High Gain” mode to increase the sensitivity for small particle detection by adjusting the CCD-camera to a higher gain setting, thus increasing the signal obtained from the small particles while minimizing the noise.  We will present data from analyzing recombinant EVs expressing green fluorescent protein (GFP) on the ImageStream® system using “High Gain” mode.  Data from EV size measurements, GFP expression, and tetraspanin expression in calibrated units will be presented. The Cytek® Aurora™ and Northern Lights™ full spectrum flow cytometers can be upgraded with the Enhanced Small Particle™ (ESP™) detection option. With the blue SSC detector and new violet SSC collection optics that enhance small particle sensitivity and resolution, both systems allow full SSC dynamic range to detect large cells and subcellular EVs. We will show bead and EV data from our high-sensitivity detectors, flat-top beam profiling, and low-noise electronics on the Cytek® Aurora™ system and how they can be combined for small particle detection. The data presented illustrates how the various tools and technologies available from Cytek for EV analysis provide robust and reproducible results for advancing discovery in this field.