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104 protocols using su6600

1

Analyzing Itokawa Regolith Particles

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We examined surface features of 10 regolith particles (average diameters = 68–241 μm) including iron sulfide (Supplementary Table. 1) using a field emission SEM (FE-SEM; Hitachi SU6600) at the ESCuC after the initial routine description. The particles were placed on an Au-coated holder for SEM observations, without a conductive coating, using an electrostatically controlled micromanipulation system. Until these SEM observation, the Itokawa particles had never been exposed to an atmospheric environment, thus minimizing the contamination and alteration of the studied particles66 (link). We performed secondary electron (SE) imaging at accelerating voltages of 1.5 and/or 2.0 kV under high vacuum with an electron beam current of ~10 pA.
Three Itokawa particles (RA-QD02-0286, RA-QD02-0292, and RA-QD02-0325) were transferred onto an adhesive carbon-conductive tape for further analysis. We determined the elemental compositions of their surfaces with an energy-dispersive X-ray spectrometer (EDX) using an FE-SEM (Hitachi SU6600) equipped with a X-MaxN 150 mm2 (Oxford Instruments) in JAXA and an FE-SEM (Hitachi SU6600) equipped with a Bruker XFlash® FlatQUAD detector at the Institute for Molecular Science (IMS, Higashi-Okazaki, Japan). The accelerating voltage for SE imaging was 1.5 kV, whereas for EDS analysis we used 5 and 10 kV.
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2

Scanning Electron Microscopy Analysis of Atelocollagen Microstructure

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Atelocollagen samples were fixed with 2% glutaraldehyde, sequentially dehydrated in ethanol, and embedded in tert-butyl alcohol. The samples were then freeze-dried and sputter-coated with osmium using an osmium coater (Neoc-STB; Meiwafosis Co., Ltd.). Images were obtained using a scanning electron microscope (SU6600; Hitachi High-Technologies). The micro-roughness of the samples was assessed via ISO 25178-compliant motif analysis (bottom detection) using MountainsMap® software (Digital Surf) based on the SEM images. The method, presently called segmentation, is based on the application of a watershed algorithm associated with an algorithm for simplifying graphs that describe the relationships between individual points (Wolf pruning) (22 (link)). Details on feature parameters were described by Blateyron (23 (link)).
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3

Dentin Disc Preparation and Analysis

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Following the last determination of the flow rate, three characteristic specimens were selected from each group of discs. After being passed through ascending grades of alcohol, the discs were immersed in hexamethyldisilazane for 10 minutes and then left on filter paper at room temperature for 1 day. Dentin discs were fractured using a dental chisel perpendicular through the surface, coated with Au, and inspected at a 15 kV acceleration voltage on the treated and fractured surfaces (SU-6600, Hitachi High-Technologies Corporation, Tokyo, Japan).
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4

Scanning Electron Microscopy Sample Preparation

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Cells were fixed in 2.5% glutaraldehyde for 12 h at 4 °C, washed in 0.1 M PB, and postfixed in 1% OsO4 for 2 h at 4 °C. The cells were dehydrated with a series of increasing concentrations of ethanol (50%, 70%, 80%, 90%, 95% and 100%), freeze-dried with 100% t-butyric alcohol (VFD-21S, vacuum device), and coated with platinum-palladium by a heavy metal coater (SC-701, Sanyu Electron). The samples were analysed using field emission scanning electron microscopy (SU6600 Hitachi High-Technologies) with 7.0 keV acceleration.
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5

Electron Microscopy of Enterococcus Phage Infection

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Enterococcus gallinarum were infected with 10 multiplicity of infection (MOI) of phiEG37k for the indicated time periods in LB media and fixed with 2% paraformaldehyde and 2.5% glutaraldehyde in PBS, followed by post fixation with 1% OsO4 in the same buffer. The fixed samples, which were placed on poly-L-lysine-coated coverslips, were washed three times with PBS, dehydrated in graded acetone solutions (50, 70, 90, 95, 99.5%), and then immersed in tert-butyl alcohol. After replacing the ethanol with tert-butyl alcohol, the samples were dried by using a freeze-drying device (VFD-21S; VACUUM DEVICE), and then coated with osmium by using a plasma coating device (Neoc; Meiwafosis). The samples were visualized with SEM (SU6600, Hitachi High-Technologies, Tokyo, Japan).
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6

Characterization of Lignin Aggregates

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Lignin aggregates
in DMF and chloroform solutions were evaluated by a dynamic light
scattering (DLS) method using a ζ-potential and a particle size
analyzer (ELSZ2000ZS; Otsuka Electronics Co., Ltd., Osaka, Japan).
In addition, each solution was cast onto a glass coverslip and dried
in air for 1 week to remove the solvent prior to conducting scanning
electron microscopy (SEM) and Fourier transform infrared (FT-IR) spectroscopy
analysis. SEM observations were performed using an SU-6600 instrument
(Hitachi High-Technologies Corporation, Tokyo, Japan). Samples were
placed on an SEM stub, coated in gold, and then analyzed at an accelerating
voltage of 5 kV. FT-IR spectra were recorded in the attenuated total
reflectance (ATR) mode (IRAffinity-1S instrument, Shimadzu Co., Kyoto,
Japan).
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7

Membrane Microstructure Analysis by SEM

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The surface microstructure of membrane was examined by SEM (FESEM, SU6600, Hitachi High Technologies America, Inc.). Before SEM observation, the sample was dehydrated by passing the specimens through a graded series of ethanol-water mixtures, and then dried by the critical-point method. After drying the sample was sputter-coated with gold, and examined under an SEM. Two areas were scanned, (i) Mid membrane region and (ii) the junction between the red part and the yellow part of the fibrin clot (buffy coat area).
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8

Fabrication and Characterization of Nanoporous Gold Substrates

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An NPG substrate was fabricated by radio frequency (RF) sputtering and dealloying (chemical corrosion of silver by nitric acid) as follows: first, a 1000-nm thick pure gold film (>99.9 mass%) was sputtered on a Φ = 22 mm micro cover glass (Matsunami Glass Ind., Ltd., Osaka, Japan) with a RF sputtering apparatus SVC-700RF (Sanyu Electron Co., Ltd., Tokyo, Japan). Next, a 300-nm thick, gold–silver alloy (atom ratio of gold:silver = 3:7) was sputtered on the gold thin film. The alloy was then immersed in nitric acid whose concentration was 70% and washed with DPBS (Nacalai Tesque) to fabricate an NPG substrate. Two ways were employed to change the pore sizes of the NPG substrates: one was to change the conditions of dealloying and the other was to change the conditions of heat treatment after dealloying (Table S1), because it was difficult to control the pore sizes with only one way. The nanostructures of the NPG substrates were observed using a scanning electron microscope (SEM, SU-6600, Hitachi High Technologies, Tokyo, Japan). The quantitative chemical composition of the NPG substrates was analyzed with energy dispersive X-ray spectroscopy (EDXS). Five areas from three different NPG samples were randomly selected to quantify the pore size and ligament size using ImageJ software.
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9

Scanning Electron Microscopy of Amphipod Exoskeleton

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The freeze-dried amphipod sample was set on the stage of a scanning electron microscope, which was covered with a silicon plate and carbon tape to avoid energy-dispersive X-ray spectroscopy (EDS) signals from the stage. The exoskeleton of the amphipod was observed with scanning electron microscopy (SEM) (SU6600, Hitachi High-Technologies Co., Tokyo, Japan) under an accelerating voltage of 20 kV, and the elementary components were analyzed by EDS (X-MaxN, Oxford).
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10

SEM Analysis of Hydrogel Microstructure

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Following the method of Lin et al. [47 (link)], the morphology and microstructure of the gel were observed by SEM (SU6600, Hitachi High Technologies Corp., Tokyo, Japan). The gel was cut into small 2 × 5 mm2 pieces with a sharp, double-sided blade and fixed in 0.1 M sodium phosphate buffer (pH 6.8) containing 2.5 M glutaraldehyde for 4 h in a refrigerator at 4 °C overnight. After a series of rinses, the gel sample is lyophilized. A metal film was then applied to the gel using an ion-sputtering coater (E-1010, HITACHI). The images were then observed at an accelerated voltage of 5.0 kV, and the XT microscope control version 2.5.0.3 software was used to collect the images at 1000× magnification.
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