Tuesday, 3:00 PM – 5:30 PM
By Jochen Behrends, Head Fluorescence Cytometry, Research Center Borstel and Anne Gompf, Head of Flow Cytometry Core Facility,
Technische Universität Dresden
Abstract
The analysis of small particles – such as extracellular vesicles (EVs), viruses, and nanoparticles – by flow cytometry remains technically challenging due to their small size, low refractive index, and limited signal generation. However, advances in instrumentation, calibration strategies, and reference materials have improved the detection and characterization of particles in the submicron range.
This tutorial provides an overview of the theoretical and practical aspects of measuring small particles by flow cytometry. The first part introduces key theoretical foundations, including light scattering, fluorescence detection, instrument sensitivity, and methodological limitations. It summarizes insights from state-of-the-art literature and current recommendations or guidelines in the field. Practical experience from our own work with small particle analysis will also be shared, highlighting common pitfalls, controls, and calibration approaches.
In the second part, practical strategies for optimizing small particle measurements will be presented, including instrument setup, thresholding strategies, sample preparation, and the use of appropriate reference materials.
The theoretical session is open to all participants. Due to technical limitations, the subsequent hands-on training at the flow cytometers (e.g. ImageStream, Discover S8, …) is limited to 18 participants. During this session, attendees will gain practical experience in instrument setup, optimization, and data acquisition for small particle analysis.
The tutorial aims to bridge theory and practice and provide guidance to improve the reliability and reproducibility of small particle measurements by flow cytometry.