Mean particle size and polydispersity index (PDI) of the nanoparticles were measured at Assuit International Center of Nanoparticle (AICN), using a Zetasizer Nano ZS instrument (Malvern Instruments, Worcestershire, UK) equipped with a backscattered light detector operating at 173°. The Zeta-potential values were measured by laser Doppler anemometry using Malvern Zetasizer Nano series ZS. All samples were diluted in distilled water and measured at 25 °C in triplicates (equilibrium time of 120 s and 15 runs). The sample volume used for all measurements was kept constant.
Zetasizer nanoseries zs
Manufactured by Malvern Panalytical
Sourced in United Kingdom, Germany
The Zetasizer Nanoseries ZS is a versatile laboratory instrument designed for the measurement of particle size, zeta potential, and molecular weight. It utilizes dynamic light scattering (DLS) and electrophoretic light scattering (ELS) techniques to provide accurate and reliable data on the physical characteristics of materials in suspension or solution.
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Lab products found in correlation
Zetasizer nano z, by Malvern Panalytical (6 mentions)
Moticam 2, by Motic (2 mentions)
Lhs h100p 1, by Nikon (2 mentions)
Zetasizer nano zs instrument, by Malvern Panalytical (1 mentions)
400 mesh carbon coated copper grid, by Electron Microscopy Sciences (1 mentions)
Dipalmitoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
Centrifugal filter unit, by Merck Group (1 mentions)
Millipore filter, by Merck Group (1 mentions)
Folded capillary zeta cell, by Malvern Panalytical (1 mentions)
Zetasizer 1000hsa, by Malvern Panalytical (1 mentions)
31 protocols using zetasizer nanoseries zs
1
Nanoparticle Characterization Protocol
2
Nanoparticle Characterization and Stability
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The mean hydrodynamic diameters and size distribution (polydispersity index [PDI]) of the nanoparticles were measured using a Zetasizer Nano ZS instrument (Malvern Instruments, Worcestershire, UK) equipped with a backscattered light detector operating at 173° as described previously. The zeta potential values were determined by laser Doppler anemometry using Malvern Zetasizer Nanoseries ZS. All measurements were performed in triplicates. The effect of different concentrations of NaCl (0.1–2 M) and dextrose (0.5%–10%, w/v) solutions on the mean hydrodynamic diameters and size distributions of PPE-, chitosan/Tpp- and PEI-based nanoparticles was assessed, as described previously.26 All measurements were performed in triplicates.
Morphology of the nanoparticles was studied by transmission electron microscopy (TEM). TEM images were collected on a JEOL 1200EX operating at 100 kV, and micrographs were recorded at calibrated magnifications using an SIA-15C CCD camera. Aqueous solutions of the samples (4 μL) were deposited onto carbon-coated copper grids, and after 1 min, the excess of the solution was quickly wicked away by a piece of filter paper. A drop of 1 wt% uranyl acetate was then added and allowed to stand for 30 s before excess stain was wicked away. The grids were allowed to dry in air overnight.
Morphology of the nanoparticles was studied by transmission electron microscopy (TEM). TEM images were collected on a JEOL 1200EX operating at 100 kV, and micrographs were recorded at calibrated magnifications using an SIA-15C CCD camera. Aqueous solutions of the samples (4 μL) were deposited onto carbon-coated copper grids, and after 1 min, the excess of the solution was quickly wicked away by a piece of filter paper. A drop of 1 wt% uranyl acetate was then added and allowed to stand for 30 s before excess stain was wicked away. The grids were allowed to dry in air overnight.
3
Characterization of Iron Oxide Nanoparticles
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All used fluidMAG- and nanoscreenMAG/G-particles were provided by chemicell GmbH, Germany. Spectroscopy was used to quantify the iron concentration of these iron oxide particles using phenanthroline staining. Additionally, the hydrodynamic diameter was measured by dynamic light scattering with the Zetasizer nano series ZS (Malvern Instruments, Herrenberg, Germany). Laser Doppler velocimetry was used to determine the zeta potential of the nanoparticles. All specifications and LOTs from the used SPIONs are listed in the supplement section (Table S1) .
4
pH-Dependent Zeta Potential of Lipid Emulsions
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Zeta potential measurements were performed by electrophoretic light scattering in a Zetasizer Nano Series ZS (Malvern Panalytical, Kassel, Germany). Samples were diluted 1:400 with a 2.77% glycerol solution. This glycerol concentration is comparable with the composition of the aqueous phase of Lipofundin®. The pH of the glycerol solution was set to different values by 1 M HCl or 1 M NaOH before the dilution, to adjust different pH values in the resulting emulsion. The pH of the resulting dilutions of the emulsions was also determined and these pH values were used for the following analysis. The deviation in zeta potential measurements caused by the changed viscosity of a 2.77% glycerol solution compared to the viscosity of water is less than 10%. Therefore, the viscosity of water at 25 °C (0.8872 mPa*s), pre-set in the measurement instrument, was used as parameter for the calculation. Each sample was diluted once and measured in three runs. In the results section, the mean values and standard deviations of the measurements are displayed.
Due to the problem with drug precipitation, no pH-zeta potential curve was recorded for BMV (cf.Section 2.4 ).
All drug concentrations given for the emulsions used for the determination of the pH-zeta potential curves represent the concentrations of the emulsions before the dilution with glycerol solution.
Due to the problem with drug precipitation, no pH-zeta potential curve was recorded for BMV (cf.
All drug concentrations given for the emulsions used for the determination of the pH-zeta potential curves represent the concentrations of the emulsions before the dilution with glycerol solution.
5
Characterization of Lipid-Based Nanoparticles
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SaNPs were characterized by dynamic light scattering (DLS) and transmission electron microscope (TEM) imaging to determine their physicochemical parameters and morphology.
Hydrodynamic size, size distribution, and zeta potential measurements were conducted using a Zetasizer Nano Series ZS (Nano-ZS, Malvern Instrument Ltd., UK) operating with a 4 mW HeNe laser (632.8 nm), a detector positioned at a scattering angle of 173°, and a temperature-controlled jacket for the cuvette. Three measurements consisting of up to 12 consecutive sub-runs were performed for each sample. Dynamic correlation functions were fitted by a second-order cumulant method to obtain the size distributions. For the zeta potential measurements, 0.8 mL of the SaNP dispersion were loaded into folded capillary zeta potential cells with integrated gold electrodes. Three measurements consisting of 12 sub-runs were performed at 25°C.
Samples for TEM were prepared by spreading 5 µl of the SaNP dispersion onto the surface of a 400-mesh carbon-coated copper grid (Electron Microscopy Sciences, USA). Uranyl acetate was used as the staining agent. The samples were air-dried at room temperature, and low-resolution TEM images were obtained using a Tecnai G2Spirit Twin T-12 electron microscope (Bar Ilan University, Israel).
Hydrodynamic size, size distribution, and zeta potential measurements were conducted using a Zetasizer Nano Series ZS (Nano-ZS, Malvern Instrument Ltd., UK) operating with a 4 mW HeNe laser (632.8 nm), a detector positioned at a scattering angle of 173°, and a temperature-controlled jacket for the cuvette. Three measurements consisting of up to 12 consecutive sub-runs were performed for each sample. Dynamic correlation functions were fitted by a second-order cumulant method to obtain the size distributions. For the zeta potential measurements, 0.8 mL of the SaNP dispersion were loaded into folded capillary zeta potential cells with integrated gold electrodes. Three measurements consisting of 12 sub-runs were performed at 25°C.
Samples for TEM were prepared by spreading 5 µl of the SaNP dispersion onto the surface of a 400-mesh carbon-coated copper grid (Electron Microscopy Sciences, USA). Uranyl acetate was used as the staining agent. The samples were air-dried at room temperature, and low-resolution TEM images were obtained using a Tecnai G2Spirit Twin T-12 electron microscope (Bar Ilan University, Israel).
6
Emulsion Particle Characterization Over Time
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During the evaporation process, an emulsion was sampled at different time intervals of 8, 16, 24, 32, 40, and 48 hours. The particle size and zeta potential of each sample were analyzed using a Zeta-sizer (Zeta-sizer nano series ZS; Malvern Instruments). Morphology changes of the particles were captured using an attachable digital C-mount camera (Moticam2; Motic®) under an inverted microscope (LHS-H100P-1; Nikon) at ×200 magnification.
7
Lipoplex Characterization by DLS and ZP
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Lipoplexes were prepared in phosphate buffered solution (PBS), for subsequent determination of size and zeta potential by dynamic light scattering (DLS) and laser Doppler anemometry, respectively, using a Zetasizer Nanoseries ZS (Malvern Instruments) by diluting the sample 50x with dd-water.
The size and size distribution of lipoplexes are indicated as displayed by the instrument. Measurement parameters were as follows: a laser wavelength of 633 nm and a scattering angle of 173° (fixed). The samples were loaded into polystyrene cuvettes and three measurements were performed, for which the mean result was recorded. Particles with PdI > 0.7 and dH > 2000 nm may be outside of the instrument's reliable range.
The size and size distribution of lipoplexes are indicated as displayed by the instrument. Measurement parameters were as follows: a laser wavelength of 633 nm and a scattering angle of 173° (fixed). The samples were loaded into polystyrene cuvettes and three measurements were performed, for which the mean result was recorded. Particles with PdI > 0.7 and dH > 2000 nm may be outside of the instrument's reliable range.
8
Intranasal Liposomal Vaccine Formulation
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Liposomal vaccines were prepared as per established methods11 (link),13 (link). Predetermined amounts of J8-palmitic acid, 3D(6-acyl) PHAD (PHAD; Avanti Polar Lipids Inc., USA) and phospholipids—dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), cholesterol (CHOL) and 1 L-α-phosphatidylglycerol (PG) (Avanti Polar Lipids Inc.) were coated on to round bottom flasks to form a lipid thin film. The thin films were hydrated in 20 mM phosphate-buffered saline (PBS; pH 6) containing trehalose and Diphtheria Toxoid (DT). The resultant liposomal suspension was freeze-dried and vaccines were stored at 4 °C until use. The average particle size (nm) of a liposome was measured using a Nanosizer (Zetasizer Nano Series ZS, Malvern Instruments) as per established method11 (link). Homogenous size distribution was confirmed by a low polydispersity index (PDI) of 0.08 for J8-Lipo-DT-PHAD with an average size of 1987 nm (SD 104 nm) before freeze-drying and PDI of 0.16 with an average size of 5213 nm (SD 572.2) after reconstitution of freeze-dried liposomes in PBS. For administration, each vaccine vial was resuspended in PBS, and mice were immunized intranasally (i.n.) with a total volume of 20 μl (30 μg/mouse of J8-palmitic acid, 25 μg/mouse of PHAD and 30 μg/mouse of DT—10 μl/nare).
9
Bovine Serum Albumin Nanoparticle Synthesis
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Nanoparticles were prepared using a desolvation method as widely reported in the literature [30] (link), [31] (link). In brief, bovine serum albumin (100 mg) was dissolved in 0.01 M Tris HCl buffer (1 mL) and the pH adjusted to 9 with 1 M NaOH. Ethanol was added drop-wise to the stirred protein (4.0 mL, 1 mL/min). Nanoparticles were cross-linked with 10% vol/vol glutaraldehyde/water (47.2 μL) and stirred overnight at room temperature, and purified into sterile PBS by four cycles of spin filtration (30 kDa molecular weight cut-off (MWCO) centrifugal filter units, Millipore, 2400 ×g, 10 min per cycle). Nanoparticle size and polydispersity were measured using a Zetasizer Nano Series ZS (Malvern Instruments, Malvern, UK). Particles were diluted 1:10 in PBS and measurements taken at 37 °C, scattering angle 173°, using viscosity value of 0.6885 cP for the dispersant. Zeta potential was measured in PBS at 25 °C at a concentration of 20 μg/mL. Particle concentration was determined gravimetrically.
10
Niosomal Dispersion Characterization
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The mean hydrodynamic diameters and size distributions (PDI) of freshly prepared niosomal dispersions in water (after 100-fold dilution and 2 min of sonication) were measured using a Zetasizer Nano ZS instrument (Malvern Instruments, Worcestershire, UK) equipped with a backscattered light detector operating at 173°. The CONTIN program was used to extract size distributions from the autocorrelation functions. The Zeta-potential values were determined by laser Doppler anemometry using Malvern Zetasizer Nanoseries ZS.
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