EV size distribution and quantification of vesicles was analyzed by Nanoparticle tracking analysis (NTA) using a NanoSight NS3000 System (Malvern Instruments, UK) Samples were suspended in 0.22 μm pre-filtered PBS and dilutions between 1:1,000 and 1:10,000 were used to achieve a particle count between 2 × 108 and 2 × 109 per mL. Measurement of the diameter was performed on 3 independent experiments and showed as mode ± standard deviation.
Nanosight ns3000 system
Manufactured by Malvern Panalytical
Sourced in United Kingdom
The NanoSight NS3000 System is a nanoparticle characterization instrument that utilizes Nanoparticle Tracking Analysis (NTA) technology to measure the size, concentration, and motion of nanoparticles in liquid samples. The system tracks the Brownian motion of individual nanoparticles and uses this information to calculate their size and concentration.
Automatically generated - may contain errors
Lab products found in correlation
Sterile pbs, by Thermo Fisher Scientific (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Dmem high glucose, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Zetasizer, by Malvern Panalytical (1 mentions)
Dts1070, by Malvern Panalytical (1 mentions)
7 protocols using nanosight ns3000 system
1
Nanoparticle Tracking Analysis of EVs
2
Nanotracking of Extracellular Vesicles
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MSC-EV samples were resuspended in sterile PBS (Thermo Fisher Scientific). Samples were diluted to a concentration suitable for further analysis. EV size distribution and quantification were analyzed on the Nanosight NS3000 System (Malvern Panalytical, Malvern, UK).
3
Nanosight Analysis of Extracellular Vesicles
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NTA was performed using the Nanosight NS3000system (Malvern Instruments, USA). XOs (either from ultracentrifugation process or controlled-release experiment) were suspended in PBS to contain ~107–1010 particles per ml, which fits within the detection limits of Nanosight NS3000. Exosomes were analyzed based on light scattering using an optical microscope aligned perpendicularly to the beam axis. A 60-s video was recorded and subsequently analyzed using NTA software.
4
Extracellular Vesicle Size Analysis via NTA
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Extracellular vesicle size distribution and quantification was performed using nanoparticle tracking analysis (NTA) on a NanoSight NS3000 System (Malvern Instruments, Malvern, United Kingdom). EV pellets were suspended in PBS in 200 μl of 0.22-μm-filtered PBS.
5
Nanoparticle Characterization in Cell Culture
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The size distribution and quantification of nanoparticles in the conditioned medium were analyzed using nanoparticle tracking analysis (NTA) with the NanoSight NS3000 system (Malvern Instruments, Malvern, Worcs, UK. The samples were suspended in serum-free RPMI medium, and dilutions were made at a ratio of 1:2 to achieve particle counts ranging from 2 × 109 to 5 × 108 per mL. Three experimental replicates were performed for each biological replicate, and two biological replicates have been conducted thus far.
6
Comprehensive Characterization of Nanosystems
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Particle size and polydispersity index (PdI) were determined by Dynamic Light Scattering (DLS), and Z-potential values by Laser Doppler Anemometry (LDA), using a Zetasizer NanoZS (Malvern Instruments, UK). Measurements were performed at 25 °C with a detection angle of 173º upon 1/10 dilution with ultrapure water (MilliQ). Nanosystems were additionally characterized by nanoparticle tracking analysis (NTA), a method to measure particle size based on imaging of individual nanosystems. Experiments were conducted with a NanoSight NS3000 System (laser operating at λ = 488 nm) (Malvern Instruments, UK). Briefly, nanosystems were injected in the sample chamber at a 1000-fold dilution in ultrapure water. Five captures, with a camera level of 14, were used to determine several parameters such as average size, homogeneity and particle concentration. Colloidal stability of the nanosystems was determined after being stored at 4 and 37 °C, as well as after incubation in biological media (DMEM high glucose, Sigma Aldrich) supplemented or not with 1% v/v fetal bovine serum (FBS, Gibco).
7
Nanocarrier Characterization by DLS, ZP, and NTA
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The nanosystems were characterized using a Zetasizer® (NanoZS Malvern Instruments, England) to determine the particle size and polydispersity index (PdI) by dynamic light scattering (DLS). All samples were analysed after dilution by 10-fold with Milli-Q water in disposable microcuvettes (ZEN0040), at room temperature, and with a detection angle of 173°. Moreover, the Z potential (ZP), indicative of the surface charge of the nanocarriers was determined using the same instrument, by Laser Doppler Anemometry (LDA) after dilution by 40-fold with Milli-Q water, in folded capillary cells cuvettes (DTS 1070, Malvern Instruments). Each sample is given by 3 measurements and the average of independent samples is provided (n = 3). Samples were also analysed by Nanoparticle Tracking Analysis (NTA) to determine the concentration of particles per millilitre and the size distribution. The measurement was carried out with a NanoSight NS3000 System (Malvern Instruments, Worcestershire, UK) equipped with a 488 nm laser and a sCMOS camera. The samples were diluted 1000-fold in ultrapure water and measured with a camera level of 14.
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