Jsm 6340f

Yeast cells were resuspended in phosphate buffer (pH 5.0) at a concentration of 30 g wet cells/L. Subsequently, 1% (w/v) PASC or Avicel was added to the cell suspension, which then was incubated at 37 °C for 2 h. Cellulose fibers were fixed with 4% paraformaldehyde and 4% glutaraldehyde (GA) in 0.1 M cacodylate buffer (pH 7.4) at 4 °C. Thereafter, fibers were fixed with 2% GA in cacodylate buffer overnight. The samples were additionally fixed with 1% tannic acid at 4 °C for 2 h. After the fixation the fibers were washed with cacodylate buffer 4 times, followed by post fixation with 2% osmium tetroxide in cacodylate buffer for 4 h. The samples next were dehydrated in a graded series of ethanol solutions (50, 70, 90, and 100%), and then were substituted into tert-butyl alcohol and dried by vacuum freeze drying. After drying, the samples were coated using an osmium plasma coater (NL-OPC80NS, Nippon Laser & Electronics Laboratory, Nagoya, Japan). The samples were visualized using a scanning electron microscope (JSM-6340F; JEOL Ltd., Tokyo, Japan) at an acceleration voltage of 5 kV. For observation using optical microscopy, post-incubation samples were directly applied onto microscope slides and imaged.

Particle and film morphology was characterized with JSM-6360 and JSM-6340F SEM (both from JEOL). Microparticles and films were coated with gold or platinum before mounting on carbon tape before imaging. The size of micron-sized particles was measured by imaging through SEM and analysis on ImageJ with a minimum of 500 particles using area-equivalent diameter. Confocal laser microscopy (CLSM) (Leica LAS) was used to visualize the extent of aggregation in PBS on particles loaded with Coumarin 6. Particles for visualization purpose was freeze-dried in 1% sucrose solution to protect against aggregation. This concentration had been shown to retain the size of particles. The particles were washed in DI water repeatedly before dispersion in PBS.
DLS for z-average hydrodynamic radius and PDI measurements were done on Zetasizer Nano ZS (Malvern Instruments). The size distribution was calculated using a built-in auto-correlation function with a backscatter detector at 173°. Size progression during coating of nanoparticles was measured in PBS for correlation with the respective release curves, unless otherwise indicated. For microparticles, the dispersant was water.

For scanning electron microscopy (SEM), eggs were fixed with 4% paraformaldehyde solution containing 1% glutaraldehyde and were then treated with 1% osmium tetroxide. The samples were dehydrated using a series of ethanol solutions and finally treated with isoamyl acetate. After the samples were subjected to critical-point drying and platinum coating (10–15 nm), the eggshells were observed with an SEM apparatus (JEOL, JSM-6340F). For transmission electron microscopy (TEM), eggs were fixed and dehydrated as described above. Then, they were treated with propylene oxide and embedded in an epoxy resin (LUVEAK-812). Ultrathin sections (90 nm) were prepared with an ultramicrotome (Leica, Ultracut), stained with uranyl acetate and lead citrate, and observed with an H-7100 (Hitachi). For thickness measurement, we obtained transverse sections of eggshells 180–200 μm and 600–620 μm from the anterior tip. The precise position of these sections was obtained from the cumulative counter of the ultramicrotome. The thicknesses of the chorion and serosal cuticle were calculated from analyses of TEM images using Photoshop (6.0, Adobe).

Each 0.2 g of BiSCaO-6, BiSCaO-2000 or SSP-Ca(OH)₂ to 100 mL of pure water, followed by rotary mixing, generated 0.2 wt% each water suspension. Then, 0.12 wt% Na₃PO₄, Na₂HPO₄, or NaH₂PO₄ was added to each suspension. The amount of H₃PO₄ were adjusted to be around pH 12. After rotary mixing, pH, average diameter, zeta potential, and form of each suspension were evaluated. Average diameter and zeta potential of particles were measured by ELSZ-1000 (Otsuka Electronics Co. Ltd., Osaka, Japan) [33 (link),34 ].
For scanning electron microscope (SEM) images of dry powder, after osmium metal coating using a neo-osmium coater (Neoc-STB; Meiwafosis Co., Ltd., Tokyo, Japan), the surface structure of each dry powder was observed with SEM images of a field-resolved scanning electron microscope (JSM-6340F; JEOL Ltd., Tokyo, Japan). For cryo-SEM, samples were frozen in liquid nitrogen, then knife-cut and observed in JEOL JSM 7100F SEM (JEOL Ltd., Tokyo, Japan) under vacuum conditions at minus 90 degrees. The accelerating voltage was 10 KV, and the detection signal was a backscattered electron image.

Based on the analysis using a scanning electron microscope (SEM), the NaF (−) and NaF (+) material surfaces were compared. All specimens were sputter-coated with Au–Pd and were assessed using SEM (JSM-6340F; JEOL, Tokyo, Japan).
The streptococci were cultured as described in the adhesion assay. After the incubation of each type of bacteria, the specimens were fixed with 1.25% glutaraldehyde (Glutaraldehyde Solution; Wako, Osaka, Japan) in phosphate buffered saline (PBS) for 2 h at room temperature. The specimens were then washed three times with PBS and dehydrated using a graded Ethanol series (70%, 80%, 90%, 95%, and 100%) (Ethanol; Wako, Osaka, Japan). The specimens were subsequently freeze-dried, sputter-coated with Au–Pd, and seven specimens from each group were assessed using SEM.

The morphologies of the obtained lignin-derived carbons were examined by the use of a field emission scanning electron microscope (FESEM, JEOL JSM 6340F, Tokyo, Japan) after coating with gold. N₂ adsorption/desorption was performed by Brunauer–Emmett–Teller (BET) measurement with a Tristar-3000 surface area analyzer (Micromeritics Instrument Corp., Norcross, GA, USA), and the specific areas were calculated by the Barrett−Joyner−Halenda (BJH) method. The lignin-derived carbon adsorbents were placed in 10 mL of RB aqueous solution with concentrations of 10 mg/L, and UV–Vis spectrophotometer (Shimadzu UV–Vis 2501PC, Tokyo, Japan) was used to measure the UV–Vis absorbance spectra of RB solution at wavelengths ranged from 200 to 700 nm. RB adsorption amounts on the adsorbents were calculated using qt = (C0 − Ct)V/m, where qt (mg/g) is the adsorption capacity; C0 is the initial concentration of RB (10 mg/L); Ct (mg/L) is the RB concentration after adsorption; V (mL) is the solution volume; and m (g) is the adsorbent dosage.