Exoviewer 3

Manufactured by Unchained Labs

The ExoViewer 3 is a laboratory instrument designed for the analysis and characterization of extracellular vesicles (exosomes, microvesicles, and other particles). It utilizes a combination of light scattering and fluorescence detection to provide detailed information about the size, concentration, and molecular composition of these nanoscale structures.

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Lab products found in correlation

Nanoviewer 2, by Unchained Labs (2 mentions) Exoview r100, by Unchained Labs (2 mentions) Exoview r100 platform, by Unchained Labs (1 mentions) Exoview platform, by Unchained Labs (1 mentions) Anti cd63, by BD (1 mentions) Anti cd9, by BioLegend (1 mentions)

3 protocols using exoviewer 3

Exosomal Tetraspanin Profiling by ExoView

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The exosome markers were analyzed using the single particle interferometric reflectance imaging sensor with the ExoView platform (NanoView Biosciences, USA). Briefly, the ExoView tetraspanin kit with immobilized antibodies against the tetraspanins CD9, CD63, and CD81 on silicon dioxide chips (NanoView Biosciences, Boston, MA) was used to capture 2D and 3D-derived EVs. Briefly, samples were diluted in PBS with 0.5% Tween-20 (PBST) and then incubated on ExoView Tetraspanin Chip for 16 h at room temperature in a 24-well plate. Chips were then washed three times in 1 ml PBST for 3 min on an orbital shaker. The chips were then incubated for 1 h at RT in a cocktail of fluorescent antibodies comprised of anti-CD9-AF647; anti-CD63-AF488; anti-CD81-AF555 diluted in 5% bovine serum albumin. After careful rinsing and drying steps, image acquisition from each chip was carried out using the ExoView® R100 platform, and the data were analyzed by the NanoViewer 2.9 along with ExoViewer 3 (NanoView Biosciences), respectively.

Jalilian E., Massoumi H., Bigit B., Amin S., Katz E.A., Guaiquil V.H., Anwar K.N., Hematti P., Rosenblatt M.I, & Djalilian A.R. (2022). Bone marrow mesenchymal stromal cells in a 3D system produce higher concentration of extracellular vesicles (EVs) with increased complexity and enhanced neuronal growth properties. Stem Cell Research & Therapy, 13, 425.

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Exosome Characterization by ExoView R100

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PFP for each animal was diluted 1:30 in separate tubes with PBS containing 0.05% Tween 20 (PBS-T). 35 μL of this sample was carefully pipetted onto the silicon chip coated with individual antibody spots against mouse CD9, CD63, and CD81 as well as negative isotype controls. After overnight incubation in a 24-well plate, chips were washed three times on an orbital shaker with PBS-T. Then, the chips were incubated for one hour at RT with a cocktail of fluorescent antibodies (anti-CD9-AF647; anti-CD63-AF488; anti-CD81-AF555) diluted in BSA (5% final concentration in well). Chips were washed once in PBS-T, three times in PBS, and once in deionized water. Chips were carefully removed from the 24-well plate, washed further in deionized water and removed for drying. Image and data acquisition for each chip was performed with the ExoView^® R100 (NanoView Biosciences). Data analysis was performed with NanoViewer 2.9 and ExoViewer 3 (NanoView Biosciences). Antibodies were purchased from BioLegend (CD9: Cat# 124802, Clone: MZ3; CD63: Cat# 143902, Clone: NVG-2; CD81: Cat# 104902, Clone: Eat-2). Fluorescent conjugation of antibodies was performed at NanoView Biosciences. A schematic illustration is shown in Figure 4A.

Khan N., Cao T., He J., Ritzel R.M., Li Y., Henry R.J., Colson C., Stoica B.A., Faden A.I, & Wu J. (2020). Spinal cord injury alters microRNA and CD81+ exosome levels in plasma extracellular nanoparticles with neuroinflammatory potential. Brain, behavior, and immunity, 92, 165-183.

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Exosome Biomarker Quantification using ExoView

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For analyzing expression of EV biomarkers and EV quantification the ExoView R100 platform (NanoView Biosciences, Boston) was used. Human ExoView Tetraspanin (EV-TETRA-C) kit was used for the analysis. The samples (EV, total sweat and patch) were processed according to the manufacturer's protocol. 1 µg of protein samples were carefully loaded onto each chip and incubated for 24 h. After that, the chips were washed three times on an orbital shaker to remove unbound particles. The chips were incubated for one hour with the human anti-CD81 (BD Pharmingen 555675), anti-CD63 (BD Pharmingen 556019), and anti-CD9 (Biolegend V P018) fluorescently labelled antibodies. Mouse IgG (Biolegend 400101) were used as controls. The immunostained chips were washed three times in PBS, once in deionized water and dried. Imaging and data acquisition of the stained chips were performed with the ExoView R100 (NanoView Biosciences) and the data analysis with the ExoViewer 3 (NanoView Biosciences).

Zhyvolozhnyi A., Samoylenko A., Bart G., Kaisanlahti A., Hekkala J., Makieieva O., Pratiwi F., Miinalainen I., Kaakinen M., Bergman U., Singh P., Nurmi T., Khosrowbadi E., Abdelrady E., Kellokumpu S., Kosamo S., Reunanen J., Röning J., Hiltunen J, & Vainio S.J. (2024). Enrichment of sweat-derived extracellular vesicles of human and bacterial origin for biomarker identification. Nanotheranostics, 8(1), 48-63.

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