Average size and population spread (polydispersity index) of liposome preparations were determined using dynamic light scattering (Nanosizer ZS, Malvern Instruments, UK). The particle size and distribution were measured after each freeze-Thaw cycle during preparation and after each week for one month during storage for measuring in vitro stability. Liposomal charge (Zeta potential) was determined using laser Doppler anemometry after a 5-fold dilution in distilled water (Nanosizer ZS, Malvern Instruments, UK). All measurements were performed in triplicate. Liposomal surface morphology was studied using transmission electron microscopy (JOEL JEM 2000 EX200) operating at an accelerating voltage of 80 kV. Samples were prepared by placing a suspension on a Formvar-coated grid and coating with a carbon layer (20 nm) under vacuum before scanning.
Nanosizer zs
Manufactured by Malvern Panalytical
Sourced in United Kingdom
The Nanosizer ZS is a dynamic light scattering (DLS) instrument used for the measurement of particle size and zeta potential of samples in liquid dispersion. The instrument determines the hydrodynamic size of particles and the surface charge (zeta potential) using non-invasive back-scatter technology.
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Lab products found in correlation
Zen0040, by Malvern Panalytical (2 mentions)
Escalab 250 spectrometer, by Thermo Fisher Scientific (2 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (2 mentions)
Optima 8000, by PerkinElmer (1 mentions)
Nicolet is50 spectrometer, by Thermo Fisher Scientific (1 mentions)
X pert pro mpd system, by Malvern Panalytical (1 mentions)
Su8010, by Hitachi (1 mentions)
Sta449f3, by Netzsch (1 mentions)
Nanophox, by Sympatec (1 mentions)
Jem 2100, by JEOL (1 mentions)
Zetasizer nano zs90, by Malvern Panalytical (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Vivaspin 6, by Sartorius (1 mentions)
Tecnai g20, by Thermo Fisher Scientific (1 mentions)
Uh5700, by Hitachi (1 mentions)
D max 2500, by Rigaku (1 mentions)
Tem instrument, by Hitachi (1 mentions)
Dts1070, by Malvern Panalytical (1 mentions)
633 nm he ne laser, by Malvern Panalytical (1 mentions)
Tecnai tf 20 x twin, by Thermo Fisher Scientific (1 mentions)
35 protocols using nanosizer zs
1
Physicochemical Characterization of Liposomes
2
Characterization of HDL-Encapsulated CaCO3 Particles
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A 20 mL droplet of the sample was placed onto a mica sheet surface. After 1 minute, excess fluid was removed. 100 mL of deionized water was used to remove excess, non-stuck particles from the mica surface. The surface was dried using filter paper and was then mounted onto a sample holder containing carbon adhesive tab. Before imaging, the sample was coated with tungsten. The samples were then imaged with a FEI Magellan 400 SEM, operating at 2 kV, and the images were analyzed using FIJI software.
2.3.3 Size and surface charge. For size determination of the HDL-encapsulated CaCO 3 templates, suspensions in a hydro dispenser were measured by light diffraction (Bettersizer Instruments). The sizes of Au-HDL-MPs and HDL-MPs were measured by dynamic light scattering (DLS) measurements at 1731, with a 400 mW argon-helium laser, operating at a wavelength of l = 632 nm at room temperature (Malvern ZS Nanosizer). For the surface charge and stability of HDL, HDL-MPs and Au-HDL-MPs, a dip cell was inserted into the solution to measure the zeta-potential (Malvern ZS Nanosizer). The HDL, Au-HDL-MPs and HDL-MPs samples were prepared as 1 mL solutions in deionized water at pH of 2-12.
2.3.3 Size and surface charge. For size determination of the HDL-encapsulated CaCO 3 templates, suspensions in a hydro dispenser were measured by light diffraction (Bettersizer Instruments). The sizes of Au-HDL-MPs and HDL-MPs were measured by dynamic light scattering (DLS) measurements at 1731, with a 400 mW argon-helium laser, operating at a wavelength of l = 632 nm at room temperature (Malvern ZS Nanosizer). For the surface charge and stability of HDL, HDL-MPs and Au-HDL-MPs, a dip cell was inserted into the solution to measure the zeta-potential (Malvern ZS Nanosizer). The HDL, Au-HDL-MPs and HDL-MPs samples were prepared as 1 mL solutions in deionized water at pH of 2-12.
3
Characterizing CL-DNA Complexes and Nanoparticles
4
Characterization of H2L Ligand Synthesis
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The ligand H2L was synthesized according to the literature method.30 All of the other reagents and chemicals were of an analytical grade and obtained from commercial sources. Powder X-ray diffraction (PXRD) data were collected on a PANalytical X’Pert PRO MPD system (PW3040/60). Fourier transform infrared (FT-IR) measurements were conducted on a Thermo Nicolet iS50 spectrometer. Scanning electron microscopy (SEM) images were taken on a Hitachi SU8010 instrument. X-ray photoelectron spectroscopy (XPS) data were obtained with a Thermo Escalab 250 spectrometer with monochromated Al-Kα excitation. The zeta potentials were determined using dynamic light scattering (DLS) on a Malvern Instruments Nanosizer-ZS. Thermogravimetric analysis (TGA) was carried out on a Netzsch STA-449F3 thermogravimetric analyzer under a nitrogen atmosphere at a heating rate of 10 °C min–1. Simultaneous inductively coupled plasma optical emission spectrometry (ICP-OES) on a PerkinElmer Optima 8000 instrument was used to determine the metal ion concentration in aqueous solution.
5
Nanoparticle Hydrodynamic Size and Zeta Potential
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The hydrodynamic size of the nanoparticles was acquired by dynamic laser scattering with a Nanophox (Sympatec, Clausthal-Zellerfeld, Germany) operated in cross-correlation mode. For the ζ potential measurements (Nanosizer ZS, Malvern Instruments, Herrenberg, Germany), which were performed at pH 7 and 25°C with a scattering angle of 90°, particles were dispersed in aqueous solutions. The experiments were performed in triplicates and, the results were averaged.
6
Morphological and Physicochemical Characterization of Liposomes
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Surface morphology was determined by a Transmission Electron Microscope (TEM) (JEM-2100, JEOL, Tokyo, Japan). In brief, liposome samples (10 µL) were dropped onto copper 400-mesh grids. After draining via a filter paper for 30 min, a phosphotungstic acid stain solution (1.5% by weight, adjusted to pH 6.0) was applied for 10 min, and TEM images were taken. The hydrodynamic sizes and polydispersity indices (PDIs) of Mex@MLipo and Mex@Lipo were measured using a Zeta-Sizer Nano-ZS 90 (Malvern Instruments, Malvern, UK). The zeta potential of the formulations was determined after a tenfold dilution by laser Doppler velocimetry using a Nanosizer ZS with a universal dip cell (Malvern Instruments, UK). Each sample was measured in triplicate.
7
Synthesis and Characterization of Leukosomes
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Leukosomes were synthesized according to the protocols designed in our laboratory and previously described [22 (link)]. Briefly, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and cholesterol (4:3:3 molar ratio) were mixed with J774 macrophage membranes using the NanoAssemblr™ platform (Precision Nanosystem, Vancouver, BC, Canada). Cyclodextrin-complexed dexamethasone (DEX) was added to the aqueous phase during assembly. Unencapsulated free DEX was separated from the encapsulated one through ultracentrifugation at 130,000× g for 1 h at 20 °C, the supernatant was removed, and the pellet resuspended in PBS pH 7.4 for further experiments. The sizes and surface charges of the nanovesicles were characterized as previously reported [18 (link)]. Dynamic light scattering (DLS) analysis using a Nanosizer ZS (Malvern Instruments, Malvern, UK) was carried out to measure diameter and size homogeneity, respectively. Surface charge (Zeta potential) was measured using a ZetaSizer Nano ZS (Malvern Instruments, Malvern, UK). All the results are the average of at least 5 measurements, with 10 runs each.
8
Trastuzumab Aggregation Induced by pH and Salt
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Trastuzumab was obtained as a dry powder. Stock solution was prepared by dissolving the trastuzumab powder in water to an antibody concentration of 50 mg/mL. To induce aggregation, the salt concentration and pH of the trastuzumab solution was changed by diluting an aliquot of the stock solution 50 times in an aqueous solution of 20 mM sodium acetate, 50 mM sodium chloride, pH 4.6. Aggregation was then induced by heating at 75 °C for 30 min. Dynamic light scattering (DLS) was performed on the trastuzumab solution, before and after heating, to verify that aggregation occurred. The DLS measurements were performed on a Nanosizer ZS (Malvern), duplicate measurements at 173° scattering angle, measurement time of 70 s.
9
Plasma Protein Interaction with Iron Oxide Nanoparticles
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Blood was taken from 10 different male Wistar rats (from 12–16 weeks old) weighing 350–400 g. Blood samples (10 tubes of 3 mL) were centrifuged for 5 min at 800 RCF to pellet the red and white blood cells. The plasma supernatant was pooled and stored at −80 °C until used.
Iron oxide nanoparticles (300 µg) were diluted in phosphate-buffered saline (PBS) solution to a final concentration of 1 mg/mL. Pooled plasma (500 µL) was then added to each tube and samples were incubated on a rotary mixer for 10, 20 and 30 min at 37 °C. Following incubation, nanoparticles were separated magnetically and washed 3 times with 2 mL of sterile PBS and the final samples were processed for analysis. The evolution of colloidal properties was characterized by dynamic light scattering (DLS) using a Nanosizer ZS (Malvern).
Iron oxide nanoparticles (300 µg) were diluted in phosphate-buffered saline (PBS) solution to a final concentration of 1 mg/mL. Pooled plasma (500 µL) was then added to each tube and samples were incubated on a rotary mixer for 10, 20 and 30 min at 37 °C. Following incubation, nanoparticles were separated magnetically and washed 3 times with 2 mL of sterile PBS and the final samples were processed for analysis. The evolution of colloidal properties was characterized by dynamic light scattering (DLS) using a Nanosizer ZS (Malvern).
10
TMC Nanoparticle Vaccine Formulation
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Ionic gelation technique was employed for the preparation of TMC nanoparticles. A 2 mg/mL solution of TMC in 10 mM HEPES buffer was treated with 1 mL Tripolyphosphate (TPP) aqueous solution (1.7 mg/mL) drop by drop while continuous stirring. TPP, the crosslinking agent, induces ionic complexation resulting in the formation of an opalescent dispersion indicating the formation of nanoparticles. The nanoparticles were harvested by centrifugation at 15,000 rpm for 10 min on a 10 µL glycerol bed. The particles were stored at −20°C until further use. Particles were resuspended in HEPES buffer pH 7.4 and after a brief sonication, the size, polydispersity index, and zeta potential were measured by using Nanosizer ZS (Malvern Instruments, Malvern, UK). Particles were observed and analyzed by SEM and TEM imaging. For imaging sample preparation the particles were processed as described earlier by Malik et al.31 (link)
Fifty micrograms of aqueous soluble TMC nanoparticles were subcutaneously injected individually to 6–8-week old female Balb/c mice (n=8) with or without CpG ODN (20 µg/mice). A booster dose was given 2 weeks post-immunization and mice sera were collected 28 days after the immunization. The pooled sera were stored at −80°C until further use.
Fifty micrograms of aqueous soluble TMC nanoparticles were subcutaneously injected individually to 6–8-week old female Balb/c mice (n=8) with or without CpG ODN (20 µg/mice). A booster dose was given 2 weeks post-immunization and mice sera were collected 28 days after the immunization. The pooled sera were stored at −80°C until further use.
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