Nanoparticle Tracking Analysis-based Exosome Characterization

Esoxomes are attracting increasing attention of researches in life science and pharmaceutical industry. In order to be of assistance, Creative Biolabs has established the Nanosight™ platform for exosome characterization. Based on this advanced platform, we can provide not only the size and concentration analysis but also analysis of phenotyping of exosome services for customers.

Exosome Basis

The term ‘exosome’ was proposed in 1987 to describe vesicles released from multivesicular endosomes in reticulocytes. Upon release, exosome may either circulate in the extracellular space adjacent to the site of release or enter into biological fluids (e.g., plasma, urine, and cerebrospinal fluid). Recent studies have indicated that exosome carries the genetic and proteomic contents of their parent cells. Moreover, increasing evidence has demonstrated that exosomes are important mediators of cell-to-cell communication and play crucial roles in both physiological and pathophysiological processes. They have been shown to be involved in inflammation, tumorigenesis, cardiovascular diseases, and so forth. Thus, the characterization of exosome in biofluids is required.

Exosome Characterization Methods

Due to the small sizes of exosomes, their analysis and quantification are often problematic. Commonly used techniques include isolation by ultracentrifugation, purification by sucrose cushion or gradients, immunoisolation by antibodies, western blotting, absorption onto latex beads followed by flow cytometry, atomic force microscopy and electron microscopy. Flow cytometry analysis of exosomes has also been reported, but is often confounded by the typical lower detection limit of 200–300 nm in most flow cytometers, although newly developed flow cytometers can now achieve better levels of detection. Recently, nanoparticle tracking analysis (NTA) has been utilized to visualise and measure nanoparticles within the size range of 30 nm to 1,000 nm.

A graphic representation of the nanoparticle tracking analysis (NTA) principle.Fig.1 A graphic representation of the nanoparticle tracking analysis (NTA) principle. (Szatanek, 2017)

High-end Nanoparticle Tracking Analysis Technologies to Characterize Exosome

Creative Biolabs offers robust technologies to solve the exsiting problems in exosome characterization.

  • Nanoparticle tracking analysis

    Nanoparticle tracking analysis can accurately measure vesicle size and concentration. Particles move rapidly in a liquid sample under Brownian motion and act as point refractors when they pass through a laser beam. Videos can be recorded and a detailed differential particle size distribution graph can be produced using analytical software.

  • Fluorescence nanoparticle tracking analysis

    Fluorescence nanoparticle tracking analysis (fl-NTA) allows for accurate sizing, counting, and phenotyping of exosome. Fluorophores (quantum dots) attached to antibodies or other biological probes, only vesicles fluorescently labeled are tracked. There are two protocols for the analysis of exosome using fl-NTA. The first protocol describes the use of the general cell membrane dye, to label exosome and their detection by fl-NTA. The second protocol is an antibody based method, for labeling exosome, with a Qdot-conjugated antibody specific for the target exosome marker, and then quantifying marker positive exosome using fl-NTA.

Features of Our Services

  • Highly resolution of per particle
  • Particle count and concentration measurements
  • Skillful scientific team
  • Best after-sale service

With the help of our well-established technologies and experienced scientists, Creative Biolabs committed to develop Nanosight™ platform for exosome characterization services. We receive prepared exosome samples or exosome containing tissue samples (supported by our exosome isolation service). Based on your project purpose, you can choose nanoparticle tracking analysis or fl-NTA with different exosome labelling protocol. We are happy to make it accessible to all kinds of research and industrial customers. Besides, we are open to discussions. Please don’t hesitate to contact us for more information.

Reference:

  1. Szatanek, R.; et al. The methods of choice for extracellular vesicles (EVS) characterization. Int J Mol Sci. 2017, 29;18(6).
For Research Use Only. Cannot be used by patients.
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