Imaging Exosomes Derived from HUVEC Cells via CytoViva Darkfield Hyperspectral Microscopy

Exosomes represent one of the fastest growing areas of therapeutic research in life sciences. For example, exosomes isolated from stem cells have been shown to partially recapitulate the therapeutic effects of their donor cells without the drawbacks inherent to stem cell therapy. Additionally, exosomes can be used as highly effective drug delivery vectors or act as biomarkers for certain diseases.

Traditional methods to characterize the size or number of isolated exosomes include dynamic light scattering or nanoparticle tracking analysis tools. However, there is now a growing need to identify exosomes based on their cell origin, confirm drug loads added to exosomes or track their uptake in cells.

CytoViva, Inc ’s Enhanced Darkfield Hyperspectral Microscopy has emerged as an effective tool for a wide range of exosome imaging and analysis applications. This can include label free as well as fluorescently labeled exosomes. CytoViva’s technology incorporates enhanced darkfield microscopy optics that optimize the signal-to- noise scattering effect of exosomes, allowing them to be observed below 100nm in size. When combined with hyperspectral imaging, the optical spectral response is recorded in every pixel of the image. This pixel level spectral data can be analyzed to confirm the unique spectral properties of different types of exosomes and even map their presence in cell culture or other environments.

To demonstrate this capability, the figures above illustrate exosomes derived from human umbilical cord vein epithelial cells (HUVEC). Figure 1 is an enhanced darkfield hyperspectral image of isolated exosomes that are derived from HUVEC cells.

These exosomes are shown in their native state, with no fluorescence or other alteration. In Figure 2, a spectral library from individual pixels of the hyperspectral image of isolated exosomes is shown.

Figure 3 illustrates an enhanced darkfield hyperspectral image of HUVEC cells from which the exosomes in Figure 1 were originally derived.

Figure 4 illustrates spectral mapping of endosomes in the HUVEC cells. This spectral mapping utilized the spectral library from the isolated exosomes in figure 2, which were shed from the HUVEC cells. This mapping illustrates that the spectral response of the isolated exosomes is a match to the endosomal elements of the HUVEC from which they were derived.??

While this example illustrates the power of hyperspectral microscopy to characterize the origin point of isolated exosomes, it can also be used to determine drug loads on exosomes or to spectrally map exosomes that have been internalized by target cells for therapy. Additionally, if your work requires the use of fluorescent labels to identify exosomes, this technique can be used to multiplex a combination of different fluorescently labeled exosomes in a single mixed?sample.

?If you have questions regarding your exosome research and how CytoViva’s technology can advance these efforts, please contact us at [email protected]. We would be pleased to discuss your research and test image samples if appropriate.

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