Researchers from SiMPore and the University of Rochester presented two posters at the 2023 International Society for Extracellular Vesicles annual meeting which featured nanomembrane-based analysis of single extracellular vesicles. The scientific poster presented data jointly by authors from SiMPore and the University of Rochester. A second poster in the exhibition hall featured a guide for single EV analysis using SiMPore’s µSiM-EV technology.
Simplified visualization of extracellular vesicle preparations by silicon nanomembrane-enabled microfluidic devices
Neville et al, 2023, International Society for Extracellular Vesicles annual meeting.
Developing and characterizing methods for separation and concentration of extracellular vesicles (EVs) remains an area of significant interest. EV applications increasingly require isolated EVs of high purity and yield to perform basic biological, diagnostic, and especially therapeutic analyses. An ability to characterize EVs by simple and rapid means before, during, and after isolation steps would benefit understanding of both currently used methods and development of new methods. However, assessing EV preparations for EV size, concentration, and presence/absence of molecular cargo currently requires multiple dedicated instruments and practiced knowledge of these systems’ procedures and pitfalls. We aim to bridge this gap by developing a user-friendly EV visualization procedure for determining concentration and detection of molecular markers within EV preparations.
Here, we report on our assessment of a microfluidic device capable of rapid visualization of EVs using simple pipet-driven loading and on-membrane EV capture and imaging. This device, named the µSiM-EV, was assessed for its ability to catch and visualize EVs by a number of microscopy techniques. On-membrane immunofluorescence was used to detect EV surface proteins, while comparisons to nanoparticle tracking analysis were used for assessing EV concentration.
We assess nanoporous silicon nitride membranes, which are incorporated into the µSiM-EV device, for their ability to capture small- and medium-sized EVs. We showcase the ability to visualize EVs when captured on-membrane by optical and electron microscopy. We further compare nanoparticle tracking analysis to the µSiM-EV for particle concentration determination.
The µSiM-EV offers ease of use for rapid EV visualization during separation and concentration procedures. Future work will focus on expanding the variety and number of EV preparations from different sources that are analyzed via the µSiM-EV procedure.
µSiM-EV: Silicon Membrane-Enabled Microfluidics for Simplified Single Extracellular Vesicle Visualization
SiMPore Inc., 2023, International Society for Extracellular Vesicles annual meeting.
The µSiM-EV enables imaged-based analysis of individual EVs.
Works with EVs prepared by ultracentrifugation, size-exclusion chromatography, or filtration from a variety of biofluids.
Requires simple pipette-driven loading and conventional epifluorescent microscopy, with no specialized equipment needed.