The morphology of NP in suspension and after reconstitution (either unloaded or loaded) was observed by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Both formulations were washed by ultracentrifugation (using the same conditions described above) to eliminate the surfactant (PVA). The samples, for TEM measurements, were fixed in a grid, treated with uranyl acetate and then, were observed in a JEOL JEM-1400 electron microscope (JEOL Ltd, Tokyo, Japan). For SEM measurements, samples were seen in a FEI Quanta 400 FEG SEM microscope (FEI, Hillsboro, OR, USA). Before the observation, samples were mounted onto metal stubs and vacuum-coated with a layer of gold/palladium.
Jem 1400 electron microscope
Manufactured by JEOL
Sourced in Japan, United States, United Kingdom, France
The JEM-1400 is a transmission electron microscope (TEM) manufactured by JEOL. It is designed to provide high-resolution imaging capabilities for a wide range of materials and applications. The JEM-1400 utilizes an electron beam to produce magnified images of samples, allowing for the observation of fine details at the nanoscale level.
Automatically generated - may contain errors
Lab products found in correlation
Ultramicrotome, by Leica camera (13 mentions)
Zetasizer nano z, by Malvern Panalytical (6 mentions)
Leica uc6 ultramicrotome, by Leica Microsystems (5 mentions)
Orius sc1000 ccd camera, by Ametek (5 mentions)
Uranyl acetate, by Merck Group (5 mentions)
Spurr s resin, by Electron Microscopy Sciences (4 mentions)
Durcupan, by Merck Group (4 mentions)
Leica ultracut s ultramicrotome, by Leica camera (4 mentions)
Lead citrate, by Merck Group (4 mentions)
Charge coupled device camera, by Olympus (4 mentions)
Veleta 2k 2k ccd camera, by Olympus (4 mentions)
Em uc7 ultramicrotome, by Leica camera (3 mentions)
Uc7 microtome, by Leica camera (3 mentions)
Quemesa camera, by Olympus (3 mentions)
Uranyl acetate, by Ted Pella (3 mentions)
Lead citrate, by Ted Pella (3 mentions)
Tannic acid, by Electron Microscopy Sciences (3 mentions)
Vt1200 vibratome, by Leica camera (3 mentions)
Uranyl acetate, by Electron Microscopy Sciences (3 mentions)
Formvar carbon 400 mesh copper, by Agar Scientific (3 mentions)
317 protocols using jem 1400 electron microscope
1
Morphological Analysis of Nanoparticle Formulations
2
Ultrastructural Analysis of Apicomplexan Parasites
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Samples of extracellular T. gondii tachyzoites and P. berghei ookinetes, oocysts, sporozoites, and merozoites were fixed in 4% glutaraldehyde in 0.1 M phosphate buffer and processed for electron microscopy as previously described [87 (link)]. Briefly, samples were postfixed in osmium tetroxide, treated en bloc with uranyl acetate, and dehydrated and embedded in Spurr’s epoxy resin (TAAB Lab Supplies, UK). Thin sections were stained with uranyl acetate and lead citrate prior to examination in a JEOL1200EX or JEOL JEM1400 electron microscope (Jeol, UK).
3
Trolox Treatment on KO Cell Ultrastructure
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Aliquots of 5 × 107 logarithmically growing Px I-III KO cells were transferred into medium ± Trolox and incubated for 0.5, 1 and 2 hr. The cell fixation and embedding procedures were adapted from (Höög et al., 2010 (link)). Briefly, cells were fixed by 2.5% glutaraldehyde and 2% p-formaldehyde in 100 mM cacodylate buffer, pH 7.2 at 4°C overnight. The cells were centrifuged in fixative and the pellet was processed in one piece. After rinsing in buffer the samples were further fixed in 1% osmium tetroxide in cacodylate buffer, washed in water, and incubated with 1% uranyl acetate in water overnight. Dehydration was done in 10 min steps in an aceton gradient followed by Spurr resin embedding and polymerization at 60°C. The blocks were cut in 70 nm thin sections using a Leica UC6 ultramicrotome (Leica Microsystems Vienna, Austria) and collected on pioloform-coated mesh grids. The post-stained sections were imaged on a JEOL JEM-1400 electron microscope (JEOL, Tokyo, Japan) operating at 80 kV and equipped with a 4K TemCam F416 (Tietz Video and Image Processing Systems GmBH, Gautig, Germany). The analysis was performed by the Electron Microscopy Core Facility (EMCF) of Heidelberg University.
4
Alpha-Synuclein Fibrillation Assay
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The samples of free alpha-synuclein without additions, as well as its mixtures with polymers, were incubated in 250-µL glass vials with intense shaking at 37 °C for 36 h. Protein and polymer concentrations were 0.4 mg/mL (29 µM of alpha-synuclein monomers) and 5 mM (in terms of charged groups), respectively. The same buffer system as in other experiments was used. Immediately before application, the samples were diluted with pure water in the ratio of 1:1. The samples were adsorbed onto a Formvar film attached to 300-mesh copper grids and contrasted with 1% uranyl acetate aqueous solution. The specimens were observed in a JEOL JEM-1400 electron microscope (JEOL, Akishima, Japan) at 100 kV × 40,000 magnification. The length and diameter of fibrils were determined from the images using ImageJ (https://imagej.nih.gov/ij/ ) software.
5
Caco-2/HMEC-1 Cell Membrane Ultrastructure Analysis
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Caco-2/HMEC-1 cell membranes in Transwell® inserts were washed with HBSS and fixed with 2.5% v/v glutaraldehyde in phosphate buffer pH 7.4 for 1 h at room temperature, then replaced with phosphate buffer pH 7.4. Afterward, 1% w/v OsO4 was added to the cell samples and kept for 1 h at room temperature. Then, the samples were washed three times by deionized water and subsequently dehydrated by 50%, 70%, and 95% ethanol twice and four times with 100% ethanol. Next, the samples were embedded in Epon™ 812 resin, which was left to polymerize for 24 h at 60 °C. Thin slices (60 nm) were cut from each sample using Leica UC7 ultramicrotome (Leica microsystems, Wetzlar, Germany) and deposited onto copper grids. The samples were stained with 3% uranyl acetate in 50% ethanol for 5 min and washed with deionized water. The samples were left to dry and then examined using the JEOL JEM-1400 electron microscope (JEOL, Tokyo, Japan).
6
Visualizing OMVs using Transmission Electron Microscopy
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OMVs (0.5 μg/μL protein concentration) dissolved in PBS were carefully placed on a copper grid (200 mesh), and the excess liquid was removed by gently touching the grid with filter paper. The copper grid was then left in a cool, dry place for 20 min to facilitate the absorption of OMVs. OMVs absorbed on the copper grid were stained by floating the grid in 10 μL aqueous uranyl acetate (2%) for 60 s. The excess stain was removed by gently touching the grid with filter paper. The dried grid was then visualized under a transmission electron microscope (JEOL JEM-1400 electron microscope, JEOL, Ltd.) operating at 120 kV.
7
Ruthenium Red Staining for TEM
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The samples were stained and processed as previously described using ruthenium red [24 (link)]. Briefly, samples were stained with ruthenium red (Sigma-Aldrich, Steinheim, Germany) while in 1% glutaraldehyde, then further processed with the rOTO protocol before embedding in Epon (Serva, Heidelberg, Germany). In addition, 70 nm ultrathin sections of the tissue were analyzed using the JEOL JEM-1400 electron microscope (JEOL, Tokyo, Japan) operating at 80 kV and equipped with a 4K TemCam F416 CMOS camera (Tietz Video and Image Processing Systems GmBH, Gautig).
8
Ultrastructural Brain Tissue Analysis
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The part of brain from mice were first fixed overnight at 4 °C in PBS buffer with 2.5% glutaraldehyde. After washing with PBS buffer (0.1 M) for three times, the brain tissues were fixed at room temperature with 1% osmium-containing PBS buffer for 2 h. Subsequently, all the brain tissues were washed three times with PBS buffer and dehydrated with a graded series of acetone (50, 70, 80, 90, 95, 100%) for 15 min for each step. After infiltrated with a graded of series of mixtures (acetone/EPON 812 resin: 2/1, 1/1, 1/2) at room temperature for 1 h, pure resin was added and incubated overnight at 4 °C. Finally, the gelatin capsules were used to cover the tissues and incubated with pure EPON 812 resin at 37, 45, and 60 °C for 24 h, respectively. The tissues were cut into ultrathin sections by a diamond knife and picked up with Formvar-coated copper grids (300 mesh). All of the sections were performed counter-staining with osmic acid (1%) for 1 h and uranyl acetate (4%) for 20 min, respectively. JEOL JEM-1400 electron microscope (JEOL, Tokyo, Japan) was used to observe tissues.
9
Gametocyte Ultrastructural Analysis
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Gametocytes activated for 6 min and 30 min were fixed in 4% glutaraldehyde in 0.1 M phosphate buffer and processed for electron microscopy as previously described 71 . Briefly, samples were post fixed in osmium tetroxide, treated en bloc with uranyl acetate, dehydrated and embedded in Spurr's epoxy resin. Thin sections were stained with uranyl acetate and lead citrate prior to examination in a JEOL JEM-1400 electron microscope (JEOL Ltd, UK)
10
Ultrastructural Analysis of K562 Cells
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K562 cells, which are cell lines originated from human chronic myelogenous leukemia, were cultured in suspension at 37 C in RPMI-1640 medium (Nacalai Tesque, Kyoto) supplemented with 10% FBS (Life Technologies, Grand Island, NY) plus antibodies (Nacalai Tesque) (Hirao et al. 2018) (link).
For CF-FS, glutaraldehyde was added directly to the cell suspension to a final concentration of 2.5%. They were kept at 37 C for 10 min and stored at 4 C overnight. They were collected by centrifugation at 1400 rpm (400 G, swing rotor) for 7 min, and sandwiched between two copper discs (Fig. 1c). Likewise, for RF-FS of living cultured cells, cells were collected by centrifugation at 1400 rpm for 7 min and sandwiched between two copper discs.
The sandwiched cells were snap-frozen by plunging into melting propane cooled with liquid nitrogen. They were freeze substituted in acetone containing 2% osmium tetroxide for 2-4 days at 80 C and embedded in epoxy resin. Ultrathin sections were cut to a thickness of 50 nm with a diamond knife, picked up on single-slot grids, and placed on a Formvar film mounted on an aluminum rack (Yamaguchi and Chibana 2018) . They were stained with uranyl acetate and lead citrate (Yamaguchi et al. 2005b) , and observed in a JEM-1400 electron microscope (JEOL, Tokyo) at 100 kV as with human tissues (Fig. 1) (Yamaguchi and Chibana 2018) .
For CF-FS, glutaraldehyde was added directly to the cell suspension to a final concentration of 2.5%. They were kept at 37 C for 10 min and stored at 4 C overnight. They were collected by centrifugation at 1400 rpm (400 G, swing rotor) for 7 min, and sandwiched between two copper discs (Fig. 1c). Likewise, for RF-FS of living cultured cells, cells were collected by centrifugation at 1400 rpm for 7 min and sandwiched between two copper discs.
The sandwiched cells were snap-frozen by plunging into melting propane cooled with liquid nitrogen. They were freeze substituted in acetone containing 2% osmium tetroxide for 2-4 days at 80 C and embedded in epoxy resin. Ultrathin sections were cut to a thickness of 50 nm with a diamond knife, picked up on single-slot grids, and placed on a Formvar film mounted on an aluminum rack (Yamaguchi and Chibana 2018) . They were stained with uranyl acetate and lead citrate (Yamaguchi et al. 2005b) , and observed in a JEM-1400 electron microscope (JEOL, Tokyo) at 100 kV as with human tissues (Fig. 1) (Yamaguchi and Chibana 2018) .
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