The nanoparticle size and exosome concentration were measured with a ZetaView PMX 110 (Particle Metrix, Germany). A video was recorded with time duration of 90 s, and the rate of frames per second was 30. A laser source with a wavelength of 405 nm was used to irradiate the exosome suspension at 25 °C. The scattered light of the nanoparticles was detected, and the concentration was calculated by counting the scattered nanoparticles. Moreover, by tracking the Brownian motion trajectory of the nanoparticles, the mean-square displacement of the nanoparticles per unit time was acquired; as a result, the nanoparticle size distribution could be determined.
Zetaview pmx 110
Manufactured by Particle Metrix
Sourced in Germany, United States
The ZetaView PMX 110 is a nanoparticle tracking analysis (NTA) instrument designed to measure the size, concentration, and movement of particles in liquids. It utilizes a laser light scattering technique to visualize and analyze individual particles in real-time.
Automatically generated - may contain errors
Lab products found in correlation
Pbs buffer, by Sartorius (17 mentions)
H 7650, by Hitachi (10 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (9 mentions)
Ht7700, by Hitachi (8 mentions)
Zetaview 8, by Particle Metrix (7 mentions)
0.22 μm filter, by Merck Group (6 mentions)
Exoquick, by System Biosciences (6 mentions)
Bca protein assay kit, by Beyotime (5 mentions)
Ht7800, by Hitachi (5 mentions)
Jem 1400, by JEOL (4 mentions)
Zetaview, by Particle Metrix (4 mentions)
Optima xpn 100, by Beckman Coulter (4 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (4 mentions)
Exosome isolation kit, by Umibio (4 mentions)
Jem 1230, by JEOL (3 mentions)
Bca kit, by Beyotime (3 mentions)
Tsg101 sc 13611, by Santa Cruz Biotechnology (3 mentions)
Qiazol, by Qiagen (3 mentions)
Tsg101, by Abcam (3 mentions)
Exoquick exosome precipitation solution, by System Biosciences (3 mentions)
304 protocols using zetaview pmx 110
1
Nanoparticle Characterization via ZetaView
2
Nanoparticle Tracking Analysis of Exosomes
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Exosomes number and size were further assessed by nanoparticle tracking analysis (NTA) using the ZetaView PMX 110 (Particle Metrix, Meerbusch, Germany) and Software ZetaView 8.02.3117 (link).
3
Particle Size and Quantity Analysis
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To measure the size and quantities of isolated particles, the suspension (1×107/ml and 1×109/ml) were examined using ZetaView PMX 110 (Particle Metrix, Meerbusch, Germany) equipped with a 405 nm laser. Videos were recorded (60 s, frame rate of 30 s), and particle movement was analyzed using NTA software (ZetaView 8.02.28).
4
Exosome Size and Concentration Analysis
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Nanoparticle tracking analysis (NTA) was used to measure the concentration and size of exos by Particle Tracking Analyzer (ZetaView PMX 110, Particle Metrix, Germany). The analyzer was calibrated with polystyrene microspheres (110 nm) and washed with PBS. Each sample was analyzed at least for three times by scanning 11 detection positions each. The NTA software was used to analyze the obtained data.
5
Exosome Characterization by NTA
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Appropriate exosome samples were diluted with PBS buffer (Solarbio, Beijing, China). The particle size and concentration of exosomes were measured through nanoparticle tracking analysis (NTA) at Vivacell Biosciences using the ZetaView PMX 110 (Particle Metrix, Meerbusch, Germany) and the corresponding ZetaView 8.04.02 software. We measured and collected NTA results at a total of 11 locations. The ZetaView system was calibrated using 110 nm polystyrene particles, and the temperature was maintained between approximately 23 and 30 °C.
6
Nanoparticle Tracking Analysis of Exosomes
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A nanoparticle-tracking analysis (NTA) with a Zeta View PMX 110 instrument (Particle Metrix, Meerbusch, Germany) was used for visualization and analysis of size and count of nanoparticles in suspension. Briefly, as previously described [11 (link),23 (link),24 (link)] (Elsayed et al. 2022; Elsayed et al. 2021; Elashiry et al. 2021), data about size and concentration were acquired with ZetaView software (8.02.28) after loading 10 μL of exosome sample into the sample chamber.
7
Extracellular Vesicle Isolation and Analysis
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Extracellular vesicles were precipitated from 1ml serum using an isolation kit (miRCURY Exosome Isolation Kit, Qiagen, Venlo, Netherlands) as previously described (19 (link)) followed by extraction of cell-free RNA. For qPCR, an EV isolation and RNA extraction were performed from a second 1 ml serum aliquot. Particle concentration and size distribution were evaluated by Nanoparticle Tracking Analysis on a ZetaView PMX 110 (Particle Metrix, Meerbusch, Germany) and visualized by transmission electron microscopy. Following the Shapiro-Wilk normality test, statistical significance of particle tracking analysis data were evaluated by ordinary one-way ANOVA followed by Tukey’s multiple comparison test using Graphpad Prism (version 8.3.1) with a significance level of p ≤ 0.05. Particle concentration and size were reported as mean values ± standard deviation. Additionally, precipitated vesicles were visualized after negative staining with 4% uranyl acetate by transmission electron microscopy using a Zeiss EM900 (Carl Zeiss Microscopy GmbH, Jena, Germany) with a wide-angle dual speed 2KCCD camera at 80 kV.
8
Exosome Morphology and Sizing Analysis
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After being fixed with 2.5% glutaraldehyde, isolated exosome bullets were centrifuged at 100 000g to remove the glutaraldehyde. The bullets were negatively stained by 3% aqueous phosphotungstic acid and fixed on copper mesh Formvar grids. The morphology of exosomes samples was observed by the JEOL Transmission Electron Microscope (JEM-1230; JEOL, Tokyo, Japan). The size of exosomes was detected by nanoparticle tracking analysis (NTA), using ZetaView PMX 110 (Particle Metrix, Meerbusch, Germany) and corresponding software ZetaView 8.04.02.
9
Nanoparticle Tracking Analysis of OMVs
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Size and concentration of OMVs were estimated by nanoparticle tracking analysis using the ZetaView PMX 110 (Particle Metrix, Meerbusch, Germany) and the appurtenant software (ZetaView 8.02.28) according to manufacturer’s instructions29 (link). Results are given as an average based on five independent measurements for each OMV sample.
10
Characterization of Platelet-Derived Small Extracellular Vesicles
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The morphology of the obtained P-sEVs was determined by TEM (transmission electron microscope), and 15 μL of the P-sEV suspension was dropped onto copper grids covered with a carbon support film (Zhongjingkeyi Technology, China). The samples were air-dried for 1 min at room temperature, and then, excess fluid was removed with filter paper. The samples were negatively stained with 2% uranyl acetate for 1 min. After that, the stained samples were baked under a lamp for 10 min. Finally, TEM (FEI, USA) was performed at 200 kV to visualize and examine the morphology of the P-sEVs.
The particle size and concentration of P-sEVs were measured using nanoparticle tracking analysis (NTA) at VivaCell Shanghai with ZetaView PMX 110 (Particle Metrix, Meerbusch, Germany) and corresponding software ZetaView 8.04.02. Isolated P-sEV samples were appropriately diluted using 1X PBS buffer (Biological Industries, Israel) to measure the particle size and concentration. NTA measurements were recorded and analyzed at 11 positions. The ZetaView system was calibrated using 110 nm polystyrene particles. The temperature was maintained at approximately 23 °C.
The particle size and concentration of P-sEVs were measured using nanoparticle tracking analysis (NTA) at VivaCell Shanghai with ZetaView PMX 110 (Particle Metrix, Meerbusch, Germany) and corresponding software ZetaView 8.04.02. Isolated P-sEV samples were appropriately diluted using 1X PBS buffer (Biological Industries, Israel) to measure the particle size and concentration. NTA measurements were recorded and analyzed at 11 positions. The ZetaView system was calibrated using 110 nm polystyrene particles. The temperature was maintained at approximately 23 °C.
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