Extracellular vesicles (EVs) such as exosomes (70 nm – 160 nm in diameter) and microvesicles (100 nm – 1,000 nm diameter) can be harvested from cell-culture supernatants and from all bodily fluids. Current standard techniques to visualize, quantify, and characterize EVs are electron microscopy, nanoparticle tracking analyses, and dynamic light scattering. To further characterize and discriminate EVs, more exact high-throughput technologies to analyze their surface are highly desired. Although conventional flow cytometry is limited to measuring particles down to approximately 300 nm – 500 nm, a relatively new flow-cytometric method—called “imaging flow cytometry”—allows for the analysis of EVs smaller than 300 nm. This webinar will introduce viewers to the challenges, limitations, and pitfalls of flow cytometry-based EV analysis, and to the imaging flow cytometry methodology. Also covered will be techniques for analyzing exosomes, microvesicles, and apoptotic bodies in unprocessed samples, how imaging flow cytometry can be used to evaluate or reevaluate EV isolation techniques, and the advantages and disadvantages of using this method.
During the webinar, viewers will learn about:
- Using an eGFP-positive biological calibrator to optimize imaging flow cytometry acquisition and analysis parameters
- The importance of appropriate controls when using fluorescent dyes and antibodies
- The value that imaging flow cytometry brings to multiparametric EV surface analysis.
Questions from the live, online audience will be answered during the webinar.
The webinar will last approximately 60 minutes.
André Görgens, Ph.D.
University Hospital Essen
Dr. Görgens completed a Master’s in biology in Düsseldorf, Germany and a Ph.D. in hematopoietic stem cell biology at University Hospital Essen, Germany. He remained in Essen as a postdoctoral research fellow at the Institute for Transfusion Medicine to pursue research into exosomes/extracellular vesicles (EVs) and currently is working to unravel the role of EVs in normal hematopoiesis and hematological malignancies. Recently, he optimized and validated an imaging flow cytometry–based method for improved high-resolution EV analysis.
Moderator: Sean Sanders, Ph.D.
Dr. Sanders did his undergraduate training at the University of Cape Town, South Africa, and his Ph.D. at the University of Cambridge, UK, supported by the Wellcome Trust. Following postdoctoral training at the National Institutes of Health and Georgetown University, Dr. Sanders joined TranXenoGen, a startup biotechnology company in Massachusetts working on avian transgenics. Pursuing his parallel passion for writing and editing, Dr. Sanders joined BioTechniques as an editor, before joining Science/AAAS in 2006. Currently Dr. Sanders is the Editor for Custom Publishing for the journal Science and Program Director for Outreach.