The task of differentiating between Extracellular Vesicles (EVs) and other nanoparticles (e.g. Nanobubbles, protein aggregates, inorganic salt precipitates) typically found within a biological sample can be very challenging.
Here we report a unique technology for rapid determination of membranous particles along with an analysis of multiple EV biomarkers. In spite of the experimental sophistication, the operational costs of our instrument are kept to a minimum.
NTA is a well-established method to analyze the size & concentration of EVs with sizes ranging between 50 – 200 nm (Giebel and Helmbrecht, 2017; Soo et al. 2012). Unfortunately, the standard scattered-light methodology is not able to discriminate EVs from other ultrafine particles in the same size range (see figure 1).
To overcome the limitations of standard NTA, we developed a series of NTA instruments which are optimized not only for scatter light but also for fluorescence NTA (Melzer et al., 2019). Each NTA system is equipped with up to four excitation lasers along with their corresponding emission filters. Our multi-laser systems are driven by software that provides rapid and automated exchange between:
(i) the detection modes (scatter vs. fluorescence),
(ii) the different laser sources, and
(iii) the corresponding emission filters (See instrument page).
Our instruments make it possible to measure the size, concentration, zeta potential, and fluorescence intensity (up to four detection channels), all within a single sample volume (figure 2). Furthermore, our new patent-pending anti-bleaching technology allows the use of fast bleaching fluorophores like e.g. FITC and Calcin.
Giebel B., Helmbrecht C. Methods to Analyze EVs; Methods Mol Biol. 2017;1545:1-20.
Soo C.Y., Song Y., Zheng Y., Campbell E.C., Riches A.C., Gunn-Moore F. and Powis S.J. Nanoparticle tracking analysis monitors microvesicle and exosome secretion from immune cells. Immunology. 2012, 136:192–197. doi:10.1111/j.1365-2567.2012.03569.
Melzer C, Rehn V, Yang Y, Bähre H, von der Ohe J, Hass R. Taxol-Loaded MSC-Derived Exosomes Provide a Therapeutic Vehicle to Target Metastatic Breast Cancer and Other Carcinoma Cells. Cancers (Basel). 2019 Jun 9;11(6). pii: E798. doi: 10.3390/cancers11060798.
Techology Note Introduction In addition to using Nano Particle Tracking Analysis (NTA) to measurement the size distribution and concentration of EV samples, both Microfluidic Resistive
Along with generally accepted methods are some “tricks of the trade” such as the addition of Tween® or BSA; however, some of those additions are problematic.
Application Note Download Abstract Determination of the titer of viruses and bacteriophages is an indispensable key technology in virological research and for diagnostic purposes. Depending
Platinum Presenter Talk at ISEV 2022 in Lyon Dr. Clemens Helmbrecht NTA is a multi-parameter technology Nanoparticle Tracking Analysis (NTA) is well known for measuring
The preparation of high-value biological material, such as proteins, requires a multi-step procedure of separation and purification. Each step must be optimized to reach full efficiency of yield.
Making a great exosome research tool even better, SBI has developed ExoGlow TM -NTA, a proprietary dye that enables fluorescent analysis of only the extracellular vesicles (EVs) present in a heterogeneous sample.
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