EV sizes (z-averaged diameter) were measured by dynamic light scattering (DLS) using Zetasizer (Nano ZS; Malvern). EVs were obtained by ultracentrifugation and diluted in PBS, samples were then transferred to a disposable cuvette, and 10 measurements for each sample were performed with a refractive index of 1.33 and absorption at 0.01. Data analysis was performed using Zetasizer 7.11 software (Malvern).
Zetasizer 7
Manufactured by Malvern Panalytical
Sourced in United Kingdom
The Zetasizer 7.11 software is a tool used for particle and molecular size analysis. It provides measurements of the hydrodynamic size, zeta potential, and electrophoretic mobility of particles and molecules in a sample. The software is designed to work with Malvern Panalytical's Zetasizer instruments to provide accurate and reliable data for a variety of applications.
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Lab products found in correlation
Zetasizer nano z, by Malvern Panalytical (8 mentions)
Malvern zetasizer instrument, by Malvern Panalytical (2 mentions)
Sas version 9, by SAS Institute (2 mentions)
Malvern zetasizer nano z, by Malvern Panalytical (1 mentions)
Zetasizer nano, by Malvern Panalytical (1 mentions)
Nanosight lm10, by Malvern Panalytical (1 mentions)
Phosphate buffered saline (pbs), by Merck Group (1 mentions)
Image lab 6, by Bio-Rad (1 mentions)
Facsdiva 6, by BD (1 mentions)
Softmax pro 5, by Molecular Devices (1 mentions)
Prism 7, by GraphPad (1 mentions)
L glutamine culture medium, by Thermo Fisher Scientific (1 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Cary 60 uv vis spectrophotometer, by Agilent Technologies (1 mentions)
Zen 0040 disposable micro cuvette, by Malvern Panalytical (1 mentions)
18 protocols using zetasizer 7
1
Extracellular Vesicle Size Measurement
2
Characterizing Nanoparticle Formulations by DLS
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The intensity-weighted (Z-average) hydrodynamic diameter (Dh) and PdI of the formulations were determined by dynamic light scattering (DLS) using a PN3702 Zetasizer Nano ZS (Malvern Instruments, Malvern, UK) equipped with a He–Ne Laser at 633 nm, set on 173° backscattering mode, at 25°C. Dh and PdI were calculated by the Zetasizer 7.11 software (Malvern Instruments) using the Stokes–Einstein equation with cumulant analysis, considering values of 0.8872 cP for water viscosity and 1.330 for water refractive index. To measure the mean emulsion droplet size and size dispersity (PdI), all formulations were previously emulsified in Milli-Q water at 37°C at 1:200 dilution and vortexed for 20 s. Values reported are the mean ± standard deviation of three readings of at least five formulations.
Zeta potential analysis was performed by laser Doppler anemometry (LDA) coupled to microelectrophoresis on the same equipment used for DLS analyses. The zeta potential values were determined on samples further diluted (100-fold) in Milli-Q water in disposable folded capillary zeta cells.
Zeta potential analysis was performed by laser Doppler anemometry (LDA) coupled to microelectrophoresis on the same equipment used for DLS analyses. The zeta potential values were determined on samples further diluted (100-fold) in Milli-Q water in disposable folded capillary zeta cells.
3
Characterization of PEG-Coated Silver Nanoparticles
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PEG–AgNPs were examined using transmission electron microscopy (TEM), as previously reported [8 (link),45 (link)]. In brief, the AgNP suspensions were sonicated for 15 min at room temperature (RT). The PEG–AgNP suspensions were placed on a 200 mesh Formvar/Carbon-coated copper grid and allowed to dry at room temperature for an hour. Thereafter, the grids were analyzed, and several images were captured at various magnifications using a TecnaiTM, (Casoria, Italy) G2 Spirit transmission microscope (FEI Company, Hillsboro, OR, USA).
Regarding the zeta potential analysis of PEG–AgNPs, all samples were diluted to 10% by volume in absolute ethanol, vortexed for 10 min, and then measured for size and zeta potential. The Malvern Zetasizer instrument (Malvern Panalytical, Malvern, UK) and Zetasizer 7.11 software were used for measurement and data processing. All measurements were performed in triplicate and at room temperature.
Regarding the zeta potential analysis of PEG–AgNPs, all samples were diluted to 10% by volume in absolute ethanol, vortexed for 10 min, and then measured for size and zeta potential. The Malvern Zetasizer instrument (Malvern Panalytical, Malvern, UK) and Zetasizer 7.11 software were used for measurement and data processing. All measurements were performed in triplicate and at room temperature.
4
Dynamic Light Scattering Analysis of Nanoparticles
5
Characterization of PEG-Coated Silver Nanoparticles
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Transmission electron microscopy (TEM) of AgNPs was performed by a method described in our previous paper [10 (link)]. Briefly, the suspensions were subjected to sonication at room temperature for 15 min prior to processing for TEM. A drop of PEG-AgNPs suspensions was deposited on a 200-mesh Formvar/Carbon coated copper grid and allowed to dry for 1 h at room temperature. Then, the grids were examined and photographed at different magnifications using Tecnai™ G2 Spirit transmission microscope (FEI Company, Hillsboro, OR, USA).
Regarding zeta potential analysis, the PEG-AgNPs suspensions were diluted to 10% by volume in absolute ethanol, vortexed for 10 minutes, and then were subjected to the size distribution and zeta potential measurements using Malvern zetasizer instrument (Malvern Panalytical, UK) and Zetasizer 7.11 software for the measurement and data processing. All measurements were carried out at room temperature and were done in triplicate.
Regarding zeta potential analysis, the PEG-AgNPs suspensions were diluted to 10% by volume in absolute ethanol, vortexed for 10 minutes, and then were subjected to the size distribution and zeta potential measurements using Malvern zetasizer instrument (Malvern Panalytical, UK) and Zetasizer 7.11 software for the measurement and data processing. All measurements were carried out at room temperature and were done in triplicate.
6
Granulometric Analysis of Extracellular Vesicles
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To control the size of the isolated MVs, granulometric analysis was carried out using the dynamic light scattering method and the Zetаsizer NanoZS laser correlation spectrometer (Malvern Instruments, UK). The particles ranged from 0.3 nm to 10 μm. MV diameter was calculated using Zetasizer 7.11 software (Malvern Instruments, UK). The size of the MVs produced by NK cells of the NK-92 cell line ranged from 210-490 nm, and the peak of the MV quantity distribution was 315 nm. These data complied with our previous work [35] and that of other research groups that ascertained the size of MVs produced by various cells [20, 71, 75] . The reproducibility of the MV granulometric analysis results obtained in our laboratory at different times has led us to recommend laser correlation analysis as a standard for assessing the isolation purity of these extracellular objects.
7
Characterization of Lipid Nanoparticle Formulations
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LNP size,
polydispersity index (PDI), and zeta potential were measured using
the Zetasizer Nano (Malvern Instruments) and Zetasizer 7.12 software
(Malvern) with the following settings: material refractive index of
1.4, absorbance of 0.01, dispersant viscosity of 0.882 cP, refractive
index of 1.33, and dielectric constant of 79. LNPs were diluted 10-fold
in PBS and equilibrated at room temperature (RT) prior to analysis
in a plastic cuvette for particle size measurements or in a Dip cell
for zeta potential measurements. Three measurements of up to 100 runs
were collected for each sample until the value equilibrated.
polydispersity index (PDI), and zeta potential were measured using
the Zetasizer Nano (Malvern Instruments) and Zetasizer 7.12 software
(Malvern) with the following settings: material refractive index of
1.4, absorbance of 0.01, dispersant viscosity of 0.882 cP, refractive
index of 1.33, and dielectric constant of 79. LNPs were diluted 10-fold
in PBS and equilibrated at room temperature (RT) prior to analysis
in a plastic cuvette for particle size measurements or in a Dip cell
for zeta potential measurements. Three measurements of up to 100 runs
were collected for each sample until the value equilibrated.
8
Characterization of TYR-βCD and TYR/CS Complexes
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The measurements for size, polydispersity index (PDI) and ζ-potential were performed in a Zetasizer Nano ZS, Malvern Instruments Ltd. (Malvern, UK) using a cuvette DTS1070. The results were analysed with the Zetasizer 7.12, Malvern Instruments Ltd. Software.
ForTYR-βCD , 1 mg of the dried sample was dissolved in 4 mL of water and was fully dispersed with 2 min ultrasound at a 2210 Ultrasonic Bath, Branson. DLS measurements of the TYR/CS and TYR-βCD/CS samples were conducted by diluting 1 mL of freshly made sample in 1 mL of water.
For
9
Liposome Size Determination by Dynamic Light Scattering
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Liposome size was determined using a Zetasizer Nano-ZS and Zetasizer 7.02 software (Malvern Instruments, Malvern, UK). Liposome suspensions were diluted to a concentration of 400 μM in sucrose-Tris buffer [280 mM sucrose, 10 mM Trizma-hydrochloride (pH 7.4), and particle size recorded prior to and following 5 mM additions of F3 or F6 using dynamic light scattering. The particle diameter (Z-average) and polydispersity of the liposome population were recorded for three populations, from which the average particle diameter was calculated.
10
Preparation and Characterization of DOPC/DOPG Liposomes
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Large unilamellar vesicles (LUVs) containing DOPC/DOPG (3:2 molar ratio, 1 mM stock concentration) of a defined size were obtained from a vacuum drying lipid chloroform solution. After the chloroform evaporation, liposomes were suspended with 10 mM Tris/HCl, 20 mM NaCl, pH 7.4. The distribution and the mean hydrodynamic range of the liposomes in suspension was determined by dynamic light scattering (DLS) with a Zetasizer Nano ZS Malvern, and the data were analysed by its built-in software (Zetasizer 7.02, Malvern Panalytical, Malvern, Worcershire, UK).
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