The distribution of particle size was measured using Fiber-Optical Dynamic Light-Scattering Spectrophotometer FDLS-3000 (Otsuka Electronics, Osaka, Japan) equipped with a 532 nm diode laser. The measurement was carried out by a scattering angle of 90° at 25°C. Size distributions were determined by cumulant and histogram analysis of DLS data using the software provided by the manufacturer. The morphology and size distribution of the sCA nanoparticles was further analyzed by laser microscope (OLS-4100, Shimadzu) and atomic force microscopy using a scanning probe microscope (SPM-9500, Shimadzu) in dynamic mode and equipped with a microcantilever (OMCL-AC240TS-R3, Olympus). Optical density (OD) of the samples at 655 nm was measured spectrophotometrically at different pH using a microplate reader (680 XR, Bio-Rad). The zeta potential of the particles was measured by Laser Doppler Micro-electrophoresis (Zetasizer Nano Z, Malvern) at 25°C for 30 sec.
Spm 9500
Manufactured by Shimadzu
Sourced in Japan
The SPM-9500 is a scanning probe microscope (SPM) designed for high-resolution imaging and analysis of surface topography and properties. It is capable of operating in various modes, including atomic force microscopy (AFM) and scanning tunneling microscopy (STM). The SPM-9500 provides precise measurements of surface features at the nanometer scale.
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6 protocols using spm 9500
1
Characterization of sCA Nanoparticles
2
Comprehensive Characterization of HAP and Mg-HAP
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Surface morphologies and elemental distributions of the HAP and Mg-HAP surfaces were investigated by SEM coupled with X-ray energy dispersive spectrometry (SEM-EDS, Hitachi S-4800). Surface roughness of the prepared surfaces was analyzed in air by the AFM (Shimadzu SPM-9500). Surface charges of the HAP and Mg-HAP surfaces in deionized water were determined using SurPASS (AntonPaar). Surface hydrophilicities of the HAP and Mg-HAP surfaces were analyzed at 22 °C in air by a sessile drop method of distilled water (approximately 0.02 mL) with automatic contact angle meter (Kyowa Interface Science CA-W200). Thicknesses of the HAP and Mg-HAP coatings were examined with a spectral ellipsometer (UVISEL/VIS Jobin Yvon). Concentrations of ions released from the HAP and Mg-HAP surfaces at 37 °C for 1, 3, and 7 days were examined by inductively coupled plasma optical emission spectroscopy (ICP-OES, Optima 8300).
3
AFM Imaging of PVA-Based Composite Films
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For the AFM imaging, an SPM-9500 (Shimadzu Co. Ltd., Kyoto, Japan) and a D3000 (Digital Instruments, Santa Barbara, CA, USA) were employed. A 25 mm2 each of the neat PVA, PVA+MMt, PVA+MMt+2%CNCs, PVA+MMt+4%CNCs, PVA+MMt+6%CNCs, and PVA+MMt+8%CNCs films was glued on double-sided tape onto the sample stage. Then, a 120 m scanner from Digital Instruments and a Model TESP etched silicon probe with a resonance frequency of about 270 kHz. The imaging duration for each frame was estimated to be between 4 and 10 min, based on the scan rate range of 0.5 to 1 Hz. In order to avoid damaging the device, the set line voltage was changed to the lowest permissible voltage, between 1.6 and 2.5 V, and the images of the films were recorded at 512 × 512 pixels.
4
Plasma-Induced Surface Morphology Alteration
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Change in surface morphology, branched polymer chain, and atoms implantation occurred simultaneously on the surface of the polymer when the polymer was exposed to the plasma. Surface roughness was highly dependent on the treatment parameter in response to the etching reaction. Plasma modified and unmodified PCL and PDMS films were characterized in terms of surface roughness using atomic force microscope (AFM, SPM-9500, Shimadzu, Kyoto, Japan) to quantify the roughness variation on the micro and nanoscale. An AFM probe was used to inspect free radical forming and ion bombardment reactions on the surface of treated films caused by plasma etching reactions. The measurement was operated in tapping mode using AFM tip (RFESP-75, Veeco, Plainview, NY, USA) with a nominal spring constant of 3 N/min and approximate resonant frequency around 75 kHz in the ambient environment. The scanning area for image comparison was kept at 25.0 μm2 with a scanning rate of 0.5 Hz. The measurements were replicated five times and the data were picked up randomly at the surface of the sample.
5
Size Analysis of CA Nanoparticles
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The size distributions of CA nanoparticles and CA-CpG were analyzed with a Zetasizer Nano ZS (Malvern Instruments, Malvern, Worcestershire, UK). Specifically, the mean diameters and particle size distributions of the nanoparticles in mouse serum albumin (Sigma-Aldrich) were measured by means of a dynamic light-scattering method using capillary cells. The size distributions of both types of nanoparticles were also analyzed by atomic force microscopy conducted with a scanning probe microscope (SPM-9500, Shimadzu, Kyoto, Japan) operated in dynamic mode and equipped with a microcantilever (OMCL-AC240TS-R3, Olympus, Tokyo, Japan).
6
Morphological Analysis of CA-[Glc] Complex
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The morphology and size distribution of the CA-[Glc] complex was analysed using a scanning probe microscope (SPM-9500, Shimadzu) in dynamic mode, equipped with a microcantilever (OMCL-AC240TS-R3, Olympus). All samples were air dried for 5 min and analysed on freshly cleaved mica substrate.
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