For negative staining, vesicles were adsorbed onto glow-discharged Formvar-Carbon Niquel grids, washed with distilled water and stained with freshly prepared 2% uranyl acetate in aqueous suspension. Negative stained samples were imaged at room temperature using a JEM-1230 transmission electron microscope (JEOL) equipped with a thermionic tungsten filament and operated at an acceleration voltage of 120 kV. Images were taken using the ORIUS SC1000 (4008 × 2672 pixels) cooled slow-scan CCD camera (GATAN). For cryo-electron microscopy, EV preparations were directly adsorbed onto glow-discharged holey carbon grids (QUANTIFOIL, Germany). Grids were blotted at 95% humidity and rapidly plunged into liquid ethane with the aid of a VITROBOT (Maastricht Instruments BV, The Netherlands). Vitrified samples were imaged at liquid nitrogen temperature using a JEM-2200FS/CR transmission cryo-electron microscope (JEOL, Japan) equipped with a field emission gun and operated at an acceleration voltage of 200 kV.
Jem 1230 transmission
Manufactured by JEOL
Sourced in Japan, United States
The JEM-1230 is a transmission electron microscope (TEM) manufactured by JEOL. It is designed to provide high-quality imaging and analysis of a wide range of materials at the nanoscale level. The core function of the JEM-1230 is to produce detailed and magnified images of thin specimens by transmitting a beam of electrons through the sample.
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Lab products found in correlation
0.1 μm membrane filters, by Avantor (2 mentions)
Lkb ultracut microtome, by Leica (2 mentions)
Orius sc1000, by Ametek (1 mentions)
Glow discharged holey carbon grid, by Quantifoil (1 mentions)
Axs d8 diffractometer, by Bruker (1 mentions)
Qeuinox 55, by Bruker (1 mentions)
S 4800, by Hitachi (1 mentions)
Ultrascan 4000 ccd camera, by Ametek (1 mentions)
First light digital camera controller, by Ametek (1 mentions)
Em uc6 ultramicrotome, by Leica (1 mentions)
Ax80 optical microscope, by Olympus (1 mentions)
Gold conjugated secondary antibody, by Merck Group (1 mentions)
Anti cd63 antibody, by BD (1 mentions)
Anti calnexin, by Merck Group (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Origin 8, by OriginLab (1 mentions)
D8 advance, by Bruker (1 mentions)
X pert pro diffractometer, by Philips (1 mentions)
Morada 11 megapixel tem ccd microscope mounted camera, by Olympus (1 mentions)
Amt imaging system, by Advanced Microscopy Techniques (1 mentions)
22 protocols using jem 1230 transmission
1
Negative Staining and Cryo-EM of Extracellular Vesicles
2
Comprehensive Materials Characterization
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The FTIR spectra of the samples were recorded using an FTIR spectrometer (BRUKER QEUINOX-55) and KBr pellets. Energy dispersive X-ray spectroscopy (EDS) was obtained with a scanning electron microscope (Hitachi, S-4800, operating voltage, 5 kV, Japan). X-ray photoelectron spectroscopy (XPS) measurements were carried out using an ESCALAB 250Xi spectrometer (Al Kα ∼ 1486.6 eV). Transmission electron microscopy (TEM) images were collected on a JEM-1230 transmission electronic microscope (JEOL, model 2100). X-Ray diffraction (XRD) patterns were obtained using a Bruker AXS D8 diffractometer with a monochromatic Cu-Kα radiation source (λ = 0.15418 nm), and the scan range (2θ) was 5° to 70°.
3
TEM Imaging of DNA Origami Structures
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TEM imaging was carried out using a JEM-1230 transmission electron microscope (JEOL) operating at 80 kV. For sample preparation 5–10 µL of a solution containing (silicified) DNA origami structures were deposited on glow-discharged TEM grids (formvar/carbon-coated, 300 mesh Cu; TED Pella, Inc.) for at least 1 min, depending on sample concentration. For visualization, bare origami structures were negatively stained by briefly washing the grid with 5 µL of a 2% uranyl formate (UFO) solution followed by staining with UFO for 10−30 s. Silicified DNA origami were not stained, but washed twice with MilliQ water.
4
Ultrastructural Analysis of Tumor-MDSCs
5
Quantification of Quantum Dot Size
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A diluted solution of QDs or QD bioconjugates (0.2–0.5 μM) was dropped onto 400 Mesh copper-carbon grids and then dried at environmental conditions. Samples were stained with uranyl acetate 0.5% prior to their observation. Images were acquired using a JEM-1230 transmission electron microscope (JEOL, Peabody, MA, USA) operated at 100 kV. At least 10 different locations on the TEM grid were examined. Particle size was determine using Image J software (Bethesda, MD, USA). The diameter was calculated considering perfectly spherical shape of at least 50 different particles.
6
Visualizing MS2 Virus-Like Particles
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Samples of purified MS2 VLPs were diluted to 0.05 mg/mL, centrifuged at maximum speed for 15 min, and additionally filtered through 0.1 μm membrane filters (VWR). Samples were then applied onto hydrophilized carbon-coated copper grids (STEM Co.), negatively stained with 1% uranyl acetate, and visualized using a JEOL JEM-1230 transmission electron microscope (TEM) at 80 kV.
7
Visualizing MS2 Virus-Like Particles
8
Cell Ultrastructure Analysis Protocol
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All the treated cells were stained with Wright-Giemsa and observed under AX80 optical microscope (Olympus, Tokyo, Japan). The cellular ultrastructure was observed under a JEM1230 transmission electron microscope (JEOL, Japan). Additional details are provided in Supplementary Detailed methods.
9
Ultrastructural Analysis of Mouse Cardiac Tissue
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Freshly dissected cardiac tissues from the left ventricle of mice were dissected into pieces of approximately 1 mm2 and immediately immersed in a 2.5% glutaraldehyde solution for fixation, followed by overnight incubation at 4°C. Subsequently, they were fixed with 1% osmium tetroxide, dehydrated in acetone, and embedded in epoxy resin. Next, the samples were sectioned into 70-nm slices using an ultramicrotome, followed by double staining with 3% uranyl acetate and lead citrate. Last, the prepared samples were observed using a JEM1230 transmission electron microscope (JEOL, Japan).
10
Electron Microscopy of Exosomes
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Transmission electron microscopy was done as previously described [58 (link),59 (link)]. Exosomes were fixed in 2% paraformaldehyde and loaded onto formvar carbon coated grids. Exosomes adhered to carbon grids were PBS washed and immuno-stained with anti-CD63 antibody (BD Biosciences) or anti-Calnexin (EMD Millipore) overnight at 4°C. Grids were washed and incubated with 6 nm gold conjugated secondary antibody (Sigma-Aldrich) for 1 h at room temperature. Exosomes were then fixed with 2.5% glutaraldehyde, washed, contrasted in 1% uranyl acetate and imaged with a JEOL JEM 1230 transmission electron microscope.
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