TEM was used to observe exosome morphology (Hitachi H-7100 microscope; Hitachi High-Technologies Corporation). For exosome TEM observation, exosomes were fixed with 2.5% glutaraldehyde at 4˚C overnight. After washing, the samples were prepared by dropping 4 µl of exosome solution onto a formvar-coated copper grid (Sigma-Aldrich; Merck KGaA) for 2 min at 25˚C, negatively stained with aqueous phosphotungstic acid for 60 sec at 25˚C, and images were taken with a transmission electron microscope at 80 kV (magnification, x500,000; Hitachi H-7100 microscope; Hitachi High-Technologies Corporation). The images were observed using Image-Pro Plus (v6.0; Media Cybernetics, Inc.).
H 7100 microscope
Manufactured by Hitachi
Sourced in Japan
The H-7100 is a transmission electron microscope (TEM) manufactured by Hitachi. It is designed for high-resolution imaging and analysis of materials at the nanoscale. The H-7100 provides a stable and reliable platform for a variety of applications, including materials science, nanotechnology, and life sciences research.
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Lab products found in correlation
Exoquick tc kit, by System Biosciences (2 mentions)
Image pro plus v6, by Media Cybernetics (1 mentions)
Titan g2 chemistem cs probe, by Thermo Fisher Scientific (1 mentions)
D2 phaser x ray diffractometer, by Bruker (1 mentions)
Jem 2100f, by JEOL (1 mentions)
Optima 7300 dv, by PerkinElmer (1 mentions)
Himac cs150gxii, by Hitachi (1 mentions)
Thermogravimetric analyzer, by TA Instruments (1 mentions)
Raman spectrometer, by WITec (1 mentions)
5 protocols using h 7100 microscope
1
Exosome Morphology Characterization by TEM
2
Comprehensive Characterization of Catalysts
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TEM images were obtained using an H-7100 microscope (Hitachi) operated at an acceleration voltage of 120 kV. High-resolution TEM and EDS studies were carried out in a JEM-2100F (JEOL) and Titan G2 ChemiSTEM Cs Probe (FEI) operated at an acceleration voltage of 200 kV. The metal contents in catalysts were determined using ICP optical emission spectroscopy (PerkinElmer, Optima 7300DV) and ICP mass spectrometry (PerkinElmer, NexION 300×). XRD patterns were obtained with a D2 phaser X-ray diffractometer (Bruker). XPS was carried out using a spectrometer (Thermo Fisher Scientific) with Al Kα X-ray (1,486.6 eV) as the light source. All XPS spectra were aligned using the C1s peak at 284.8 eV as reference.
3
Exosome Isolation and Characterization
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Exosomes were prepared using sequential centrifugation according to methods described previously. The exosomes were isolated from the supernatants of CD133 + /Lin-/CD45- cells using ExoQuick-TC Kit (System Biosciences, USA) in accordance with manufacturer’s instructions. The CD133 + /Lin-/CD45- cells were cultured in a conditioned medium containing 10% exosome-free fetal bovine serum (FBS) for 48 h for the preparation of exosomes isolation, and then the supernatants of CD133 + /Lin-/CD45- cells were collected by initial centrifugation at 3,000 g for 15 min to pellet and remove the cell debris, then followed by sequential centrifugations at 13,000 g for 30 min, followed by centrifugation at 100,000 g for 60 min (Himac CS150GXII, HITACHI, Japan). Then the exosome-enriched fraction was resuspended in phosphate-buffered saline (PBS). To identify the Exos, exosomes resuspended in 0.1 mL of PBS was used for a transmission electron microscope and particle size analysis, and exosomes resuspended in 0.1 mL RIPA buffer for protein quantification and Western blotting. 0.5 × 109 exosomes were administered to cultured cells in vitro and 5 × 109/40μL to experiment rats in vivo to evaluate the function of Exos. Besides, the morphologies of exosomes were observed with a transmission electron microscope (Hitachi H-7100 microscope; HITACHI, Japan).
4
Bacterial Cell and OMV Imaging by TEM
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TEM was conducted at the Korea University Medical Research Center (https://medicine.korea.ac.kr/web/msrc/home ). The experimental procedure was in accordance with the instructions of the manufacturer. Briefly, for bacterial cell imaging, cells were grown for 18 hr in LB media. Aliquots were transferred onto carbon-coated copper grids (Formvar; Ted PELLA, Canada) and fixed with 2.5% glutaraldehyde diluted in 50 mM sodium cacodylate (pH 7.2). After washing three times with 3% saccharose, the cells were observed. For OMV imaging, aliquots of purified OMV samples were dispensed on carbon-coated copper grids. Excess liquid was discarded from the grids and the samples were negatively stained with phosphotungstic acid (3% [wt/vol]) for 5 min and then dried with filter paper. The specimens were examined using an H-7100 microscope (Hitachi, Tokyo, Japan) operating at an accelerating voltage of 75 kV and at magnifications of 40,000× for cells and 30,000× for OMVs.
5
Comprehensive Materials Characterization Protocol
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Powder X-ray diffraction (XRD) of the crystalline phase identification was carried out using a Shimadzu diffractometer (XRD6000) (Shimadzu, Tokyo, Japan) with Cu Kα (30 kV) radiation. The XRD patterns were recorded in the 2θ range of 10–80° with a scanning speed of 4°/min. Field emission scanning electron microscopy (FESEM) images were recorded with a Nova Nanosem 30 series microscope (FEI Company, Oregon, USA) using 5 kV. Internal morphologies of the samples were obtained via transmission electron microscopy (TEM) using the H-7100 microscope from Hitachi (Tokyo, Japan). Prior to the TEM measurements, the samples were dispersed in ethanol by sonication for 15 min before it was dropped on the copper grid. Textural properties of the samples were determined using nitrogen gas adsorption–desorption techniques with ASAP2000 analyzer, (Micromeritics, Georgia, GA, USA) with degassing was done at 105 °C overnight. The thermal decomposition of the product was investigated using a thermogravimetric analyzer, (TA Instruments, Delaware, DE, USA). The temperature range used for the thermal analysis was from room temperature to 1000 °C with a heating rate of 5 °C/min. Raman spectra of the CQNs were obtained using a Raman spectrometer (WITec, Ulm, Germany) with a 532 nm laser.
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