The FTIR spectra of MSN and MSN-GA were obtained using an FTIR spectrophotometer (Nicolet iS50; Thermo Fisher Scientific, Waltham, MA, USA) to confirm the successful synthesis of MSN-NH2 and MSN-GA. Malvern Zetasizer Nano ZS ZEN 3600 instrument (Malvern, UK) was employed to measure the size distribution and zeta potential of different samples. TEM (Hitachi-7500, Hitachi, Tokyo, Japan) was used to observe the morphology of MSN and MSN-GA. Differential scanning calorimetry (DSC) curves were obtained using a STA449 thermal analyzer (Netzsch, Germany) in the temperature range of 20°C–300°C at a heating rate of 10°C min−1.
Zetasizer nano zs zen3600 instrument
Manufactured by Malvern Panalytical
Sourced in United Kingdom
The Zetasizer Nano ZS ZEN3600 is a dynamic light scattering instrument designed for the measurement of particle size, zeta potential, and molecular weight. It utilizes non-invasive backscatter technology to provide rapid and accurate results for a wide range of sample types, including proteins, polymers, emulsions, and suspensions.
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5 protocols using zetasizer nano zs zen3600 instrument
1
Characterization of Mesoporous Silica Nanoparticles
2
Characterization of TiO2NP and MWCNT Nanoparticles
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TiO2NP (NM-102) and MWCNT (NM-401) were obtained from the repository of the EU Joint Research Centre (Ispra, Italy) in the frame of the EU NanoReg project. Further characterization of these materials was performed using transmission electron microscopy (TEM, JEOL JEM-2011 instrument) (Jeol LTD, Tokyo, Japan).) to determine dry size and morphology. The mean sizes were determined by measuring 100 randomly selected nanoparticles in several TEM images using the ImageJ program. Specifically, the mean size of MWCNT was calculated by measuring the width of isolated fibers. Dynamic light scattering (DLS), and laser Doppler velocimetry (LDV) methodologies (Malvern Zetasizer Nano-ZS zen3600 instrument) (Malvern, UK). were used to determine hydrodynamic size and zeta potential, respectively. For NMs dispersion, TiO2NP and MWCNT were pre-wetted in 0.5% ethanol and dispersed in 0.05% bovine serum albumin (BSA) in Milli-Q water. Afterward, the NMs were sonicated for 16 min to obtain a stock dispersion of 2.56 mg/mL, according to the NanoGenotox protocol [21 ]. For TEM and Zetasizer measurements, the stock suspension was dispersed in water and in culture medium, respectively.
3
Comprehensive Structural Characterization of Nanomaterials
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The solid MFCs were coated with gold and then observed under a scanning electron microscope (SEM, JSM-IT300LA, JEOL, Japan). The X-ray diffractometry and FTIR spectrometry were performed to explore the structure of the MFCs. A Helios NanoLab (Helios Nanolab G3 UC, FBI, U.S.A) was employed to observed the morphology of LNPs. The structure of the LNPs was analyzed by FTIR spectrometry, X-ray diffractometry, and X-ray photo-electron spectroscopy (XPS, ESCALAB 250Xi, Thermo Fisher Scientific). HSQC and 31 P NMR spectra of the LNPs were performed in a Bruker Avance 400 MHz spectrometer (Bruker GmbH, Karlsruhe, Germany). Furthermore, UV-vis spectroscopy (U-4100, Hitachi, Japan) analysis was employed to further explore the formation mechanism of the LNPs. The alkali lignin and dissolved lignin were characterized as the controls. The CQDs were observed under TEM and high-resolution TEM. Then FTIR spectrometer, XPS and Raman spectra (Renishaw Raman system model 1000 spectrometer with radiation at 633 nm) were employed to analyze the structure of the CQDs. Photoluminescence (PL) spectra were performed using a fluorescence spectrophotometer (Edinburgh FLsp920 transient, USA). The Zeta potential of the CQDs at pH 2-11 was measured via a Malvern Zetasizer Nano-ZS ZEN3600 Instrument (Malvern, United Kingdom).
4
Optimizing Liposome Preparation with Fluidic Mixing
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To preliminarily optimize the preparation conditions for LNPs, we first prepared EPC/chol liposomes with a fluidic device. A Deneb-type micromixer (KC-M-H-SUS; YMC CO., Kyoto, Japan; volume in mixing 32 μL, minimum diameter 0.2 mm × 0.2 mm (width × depth)) with a Harvard 33 Twin Syringe Pump (Harvard, Holliston, MA, USA) were used for the fluidic mixing. First, 10 mM EPC in ethanol and 10 mM chol in ethanol were diluted to a concentration of 0.1 -16.0 mM (total) with ethanol in a tube.
The PBS (-) and lipid solution were then allowed to flow through the micromixer at the indicated velocity with a fixed velocity ratio of lipid and PBS (-). The ethanol in the mixture was then diluted 10-fold with PBS (-), and the final produce characterized with Zetasizer Nano ZS ZEN3600 instrument (Malvern Instruments, Worchestershire, UK).
The PBS (-) and lipid solution were then allowed to flow through the micromixer at the indicated velocity with a fixed velocity ratio of lipid and PBS (-). The ethanol in the mixture was then diluted 10-fold with PBS (-), and the final produce characterized with Zetasizer Nano ZS ZEN3600 instrument (Malvern Instruments, Worchestershire, UK).
5
Lipid Nanoparticle Synthesis and Characterization
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A 90% t-BuOH solution containing YSK05/chol/chol-PEG 400 -amine/DMGm PEG 2k at a molar ratio of 70/30/15/1.5 were prepared at a concentration of 7.5 mM total lipid. In case of GalNAc modification, 0.25 to 0.5 mol% of trivalent GalNAc ligand was added to the above solution. These lipid solutions were mixed with 0.4 mg/mL siRNA solution to be N/P ratio of 8. LNPs were prepared by gradually adding this mixture to 20 mM Citrate buffer (pH4.0) under vigorous mixing. The resulting LNP solution were diluted with 0.1 M HEPES buffer (pH9.5) and ultrafiltrated using Vivaspin Turbo-15 (MWCO 100 kDa, Sartorius) twice for removal of t-BuOH, adjustment of pH. The size and ζ-potential of the LNPs were measured by a Zetasizer Nano ZS ZEN3600 instrument (Malvern Instruments, Worcestershire, UK). The encapsulation efficiency and total concentration of siRNA were measured by a Ribogreen assay, as described previously [30] .
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