S. suis ATCC 43765 cells were washed twice and resuspended in PBS (pH = 7.4). Various concentrations of floxuridine (0, 1 × MIC, 10 × MIC) were added to the cell suspension and incubated for 4 h. the cells were collected via centrifugation at 5000 rpm for 6 min and then resuspended in 2.5% glutaraldehyde overnight at 4 °C. The cell morphology was observed under a Hitachi Regulus 8100 SEM (Hitachi, Tokyo, Japan).
Regulus 8100 sem
Manufactured by Hitachi
Sourced in Japan
The Regulus 8100 is a Scanning Electron Microscope (SEM) manufactured by Hitachi. It is designed for high-resolution imaging and analysis of samples at the nanoscale level. The Regulus 8100 SEM provides advanced imaging capabilities, including secondary electron and backscattered electron detection, as well as energy dispersive X-ray (EDX) analysis for elemental composition.
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Lab products found in correlation
Uc7 ultrathin microtome, by Leica (1 mentions)
Jem 1400 flash tem, by JEOL (1 mentions)
Axis ultra dld spectrometer, by Shimadzu (1 mentions)
Vertex 80v spectrometer, by Bruker (1 mentions)
Jem 2100plus tem, by JEOL (1 mentions)
Escalab 250xi x ray photoelectron spectroscope xps, by Thermo Fisher Scientific (1 mentions)
Mpms squid vsm, by Quantum Design (1 mentions)
Ht7700 tem, by Hitachi (1 mentions)
10 protocols using regulus 8100 sem
1
Analyzing S. suis Morphology under Floxuridine
2
Pericarp Microstructure and Ultrastructure Analysis
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The pericarp microstructure was observed following the method of [14 (link)], with some modifications. Peels (0.5–1.0 cm2) were fixed in 2.5% pentanediol for 2 h (25 °C), and overnight at 4 °C and then fixed in 1% osmic acid for 2 h. Each sample was dehydrated with 30, 50, 70, 80, 90, 95, and 100% ethanol for 15 min, then were placed in a vacuum freeze-dryer for 24 h. The samples were sprayed with gold using a Cressington 108Auto ion sputterer, and then observed under Regulus 8100 SEM (Hitachi, Japan).
The pericarp ultrastructure was observed following the method of Zhao et al. [15 (link)], with some modifications. Peels (1 cm3) were fixed with 2.5% pentanediol over night at 4 °C, then were fixed in 1% osmium acid (prepared with 0.1 mol L−1, pH 7.5 phosphate buffer) for 7 h. Each sample was dehydrated with the same conditions as above. Different embedding agents were used to infiltrate the sample, including acetone: embedding medium 812 (v/v = 1:1, 24 h), acetone: embedding medium 812 (v/v = 1:3, 4 h) and pure embedding medium 812 (24 h). Afterwards, the samples were heated (37 °C) for 12 h, and then heating (60 °C) for 48 h. The ultrathin (70 nm thick) of the resin blocks were cut with a Leica UC7 ultra-thin microtome, and then dyed (2.6% lead citrate, 2% uranyl acetate) for 8 min, respectively. Observation was performed with JEM-1400 Flash TEM (JEOL, Tokyo, Japan).
The pericarp ultrastructure was observed following the method of Zhao et al. [15 (link)], with some modifications. Peels (1 cm3) were fixed with 2.5% pentanediol over night at 4 °C, then were fixed in 1% osmium acid (prepared with 0.1 mol L−1, pH 7.5 phosphate buffer) for 7 h. Each sample was dehydrated with the same conditions as above. Different embedding agents were used to infiltrate the sample, including acetone: embedding medium 812 (v/v = 1:1, 24 h), acetone: embedding medium 812 (v/v = 1:3, 4 h) and pure embedding medium 812 (24 h). Afterwards, the samples were heated (37 °C) for 12 h, and then heating (60 °C) for 48 h. The ultrathin (70 nm thick) of the resin blocks were cut with a Leica UC7 ultra-thin microtome, and then dyed (2.6% lead citrate, 2% uranyl acetate) for 8 min, respectively. Observation was performed with JEM-1400 Flash TEM (JEOL, Tokyo, Japan).
3
Characterization of Filtration Performance
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SEM images were examined by a Hitachi Regulus 8100 SEM working at an acceleration voltage of 5 kV. Fourier-transform infrared (FTIR) spectra were collected with a VERTEX80v spectrometer (Bruker Optik GmbH, Germany). X-ray photoelectron spectrometer (XPS) spectra were recorded on an AXIS UltraDLD spectrometer (Kratos, UK). The element content data of samples were collected with an elemental analyzer (2400 Ⅱ, Perkin Elmer, USA). A T200-Auto 3 Plus contact angle analyzer (BiolinScientific, Germany) was used to conduct the water contact angle test, in which a drop of distilled water (4 μL) from a micro syringe was automatically dispensed on the surface of sample. A digital air permeability tester (YG461E, Wenzhou Fangyuan Instrument Co., Ltd, China) (Fig. S2 ) was used to evaluate the air permeability. An Automated Filter Tester 8130 (TSI Inc., USA) (Fig. S3 ) was used to measure the filtration efficiency and pressure drop with a flow rate of 85 L min−1, and sodium chloride (NaCl) monodisperse aerosols with a mass median diameter of 0.26 μm were used as the model particles.
4
SEM Analysis of Aspergillus flavus Conidia
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For scanning electron microscopy (SEM) analysis, 1 × 106 conidia of A. flavus were harvested by centrifugation at 3000× g and washed with PBS (phosphate buffered saline, pH 7.4) twice. Then, conidia were fixed in 2.5% glutaraldehyde in PBS for 2 h at room temperature. The conidia were washed with PBS for 3 times, 15 min each, and then the conidia were post-fixed in 1% osmium tetroxide for 1–2 h at room temperature. After that, conidia were washed in PBS for 3 times. The dehydration of samples was achieved by transferring by increasing the concentration of (30–100%) ethanol solutions, and the samples were dried with Critical Point Dryer [39 (link)]. The samples were then attached to metallic stubs using stickers and sputter-coated with gold for 30 s. The observations were made on a HITACHI Regulus 8100 SEM (Tokyo, Japan).
5
Tomato Peel Ultrastructure Analysis
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Following previous methods [49 (link)] and modifying them, we rinsed the sample gently with distilled water to remove surface dirt and wiped it clean with a clean paper towel. Then, a clean and sterile scalpel was used to cut tomato peels (0.5 ~ 1.0 cm2) at the equator, and attention was given to minimize pulling during sampling to avoid mechanical damage and affect the observation results. The tomato skins were soaked with the fixative solution under an electron microscope and then fixed at room temperature, rinsed with 0.1 M PBS buffer (pH 7.4) three times for 15 min each to remove the fixative solution. The samples were immersed in 30%–50%–70%–80%–90%–95%–100%–100% ethanol for dehydration for 15 min each time and then dried with a vacuum freeze dryer for two hours. A Cressington 108 Autoion sputterer was used to apply gold to the samples. Next, the samples were examined using a Regulus 8100 SEM (Hitachi, Japan). 3.0 kV was set as the acceleration voltage.
6
Comprehensive Material Characterization Techniques
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Scanning electron microscopy (SEM) was performed with a regulus 8100 SEM (Hi-tachi, Tokyo, Japan). Transmission electron microscopy (TEM) images were obtained with a JEM 2100 PLUS TEM (JEOL, Tokyo, Japan). Brunauer-Emmett-Teller (BET) surface areas were measured by a Micromeritics ASAP 2460 machine (Micromeritics Corporate, Norcross, GA, USA). Fourier-transform infrared (FT-IR) spectra were measured using a Nocolet Nexus 710 (Bruker, Rheinstetten, Germany). The composition and chemical and electronic states of the elements in the Fe3O4@N-CMP were measured using an Escalab 250 Xi X-ray photoelectron spectroscope (XPS) (Thermo Scientific, Waltham, MA, USA). Magnetization curves were calculated by a vibrating sample magnetometer (Mpms Squid Vsm, Quantum Design, San Diego, CA, USA).
7
Ultrastructural Analysis of Autophagic Processes
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TEM was utilized to examine the autophagosome or autophagolysosome.18 Briefly, tissue or cell samples were fixed, post‐fixed, dehydrated, infiltrated in Epon812 and immersed in resin. The obtained ultrathin slices were then subjected to uranyl acetate and lead citrate staining and visualized and photographed utilizing a HITACHI HT7700 TEM. SEM was used to observe pyroptosis; samples were washed, fixed, dehydrated and treated with isoamyl acetate:ethanol (1:1) and isoamyl acetate for 10 min each and dried with a critical‐point drier. Afterward, the specimens were introduced into an ion sputter coater, where they underwent gold sputtering and were subsequently examined utilizing a HITACHI Regulus 8100 SEM.
8
Sequential Extraction and Characterization of Sediment Phosphorus
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The chemical sequential extraction method [26 ] was used to gradually extract soluble phosphorus (SP), aluminum-bound phosphorus (Al-P), iron-bound phosphorus (Fe-P), calcium-bound phosphorus (Ca-P), occluded phosphorus (Oc-P) and organic phosphorus (OP). NH4Cl, NH4F, NaOH, H2SO4 and other reagents were used. Oscillation, centrifugation, heating, and other operations were conducted in the overall extraction procedure. Molybdenum antimony anti-spectrophotometry was carried out to determine the concentration of phosphorus via UV spectrophotometry (TU-1950, PERSEE, Beijing, China).
To examine the intrinsic mechanism of the release and migration of phosphorus, the organic matter content, specific surface area, particle size and elemental composition of the sediment samples were determined. The organic matter content was determined by the potassium dichromate-sulfuric acid digestion method [27 ]. The specific surface area of the sediment particles was determined by nitrogen adsorption using an automatic specific surface area and porosity analyzer. The particle size and elemental composition were obtained by scanning electron microscopy and energy dispersive spectroscopy (SEM-EDS) (REGULUS8100 SEM, HITACHI, Tokyo, Japan; ASAP2460 BET, Micromeritics Instrument, Atlanta, GA, USA).
To examine the intrinsic mechanism of the release and migration of phosphorus, the organic matter content, specific surface area, particle size and elemental composition of the sediment samples were determined. The organic matter content was determined by the potassium dichromate-sulfuric acid digestion method [27 ]. The specific surface area of the sediment particles was determined by nitrogen adsorption using an automatic specific surface area and porosity analyzer. The particle size and elemental composition were obtained by scanning electron microscopy and energy dispersive spectroscopy (SEM-EDS) (REGULUS8100 SEM, HITACHI, Tokyo, Japan; ASAP2460 BET, Micromeritics Instrument, Atlanta, GA, USA).
9
GBS Dehydration and SEM Imaging
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9.6 × 108 CFU of GBS was suspended in 1 ml PBS or PBS containing TBP-1 (10 µM) at 30°C, 200 rpm, 10 min. Afterward, the GBS suspension was kept in the dark or under white light (4.2 mW cm−2) for 10 and 30 min. Subsequently, 30, 50, 70, 80, 90, 95, and 100% ethanol were used for successive dehydration of GBS fixed by glutaraldehyde (2.5%). The GBS samples were coated with gold and then measured by SEM (Regulus 8100 SEM, Hitachi).
10
Visualizing Cardiomyocyte Pyroptosis with SEM
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To facilitate the observation of cardiomyocyte pyroptosis using a scanning electron microscope (SEM), the H9C2 cardiomyocytes were challenged to 12 h stimulation with lipopolysaccharide (LPS). In brief, the cells were subjected to fixation in a 2.5% glutaraldehyde solution for 72 h. The cells were then washed and fixed in a 1% osmic acid solution at 4 ℃ for 2 h. The cells were finally imaged using a HITACHI Regulus 8100 SEM. Following drying using a critical point drier and applying a coating using an ion sputtering device, the specimens were subsequently positioned on the stage of an SEM, transported into a designated sample chamber, and captured using an HITACHI Regulus 8100 SEM.
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