Additional corneal specimens were fixed with 2.5% glutaraldehyde in Sorense-Gomori’s PBS (pH 7.4) for 2 h at 4 °C. After being washed three times with PBS, specimens were post-fixed using 1% osmium tetroxide (Wako Pure Chemical Industries, Tokyo, Japan) for 2 h at 0 °C, dehydrated in ethanol, and embedded in epoxy resin. Ultrathin sections of 70 nm were cut using an ultra-microtome (Leica UC6/FC6, Leica Microsystems, Wetzlar, Germany) equipped with a diamond knife, and mounted on grids. Sections were observed by TEM with accelerating voltage at 200 kV (LEO 922 Omega, Carl Zeiss, Germany).
Osmium tetroxide
Manufactured by Fujifilm
Sourced in Japan
Osmium tetroxide is a chemical compound used in laboratory settings. It is a strong oxidizing agent and is primarily utilized as a fixative and staining agent in electron microscopy to enhance the contrast of biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Glutaraldehyde, by Fujifilm (3 mentions)
Propylene oxide, by Nacalai Tesque (2 mentions)
Leica uc6 fc6, by Leica Microsystems (1 mentions)
Leo 922 omega, by Zeiss (1 mentions)
Ethanol, by Fujifilm (1 mentions)
Ultracut s, by Leica Microsystems (1 mentions)
Jsm 7800f, by JEOL (1 mentions)
Epoxy resin, by TAAB (1 mentions)
S 4700, by Hitachi (1 mentions)
Jsm 6701f, by JEOL (1 mentions)
Epoxy resin, by Nacalai Tesque (1 mentions)
H 7650 transmission electron microscope, by Hitachi (1 mentions)
Jcm 5700, by JEOL (1 mentions)
6 protocols using osmium tetroxide
1
Ultrastructural Analysis of Corneal Tissue
2
Epithelial Keratinization Evaluation via SEM
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As mentioned above, the non-keratinizing epithelium enhances the
keratinization of epithelium via the dehydration. To confirm that
no keratinization occurred during the aerosol exposure, SEM observations
of the EpiOral tissue after the aerosol exposure were performed. After
fixation in glutaraldehyde (FUJIFILM Wako Pure Chemical Corporation),
the specimens were rinsed in 0.2 M phosphate buffer and post-fixed
with 1% osmium tetroxide (FUJIFILM Wako Pure Chemical Corporation).
After rinsing, the specimens were dehydrated in ethanol (FUJIFILM
Wako Pure Chemical Corporation) and embedded in propylene oxide (Nisshin
EM Co. Ltd., Tokyo, Japan) and epoxy resin (TAAB Laboratories Equipment
Ltd., Berks, England). Thereafter, the tissues were cut into the cross-sectional
specimens by a microtome (Ultracuts, Leica Microsystems, Wetzlar,
Germany). The tissues were determined by the SEM (JSM-7800F, JEOL,
Tokyo, Japan) at an accelerating voltage of 5 kV.
keratinization of epithelium via the dehydration. To confirm that
no keratinization occurred during the aerosol exposure, SEM observations
of the EpiOral tissue after the aerosol exposure were performed. After
fixation in glutaraldehyde (FUJIFILM Wako Pure Chemical Corporation),
the specimens were rinsed in 0.2 M phosphate buffer and post-fixed
with 1% osmium tetroxide (FUJIFILM Wako Pure Chemical Corporation).
After rinsing, the specimens were dehydrated in ethanol (FUJIFILM
Wako Pure Chemical Corporation) and embedded in propylene oxide (Nisshin
EM Co. Ltd., Tokyo, Japan) and epoxy resin (TAAB Laboratories Equipment
Ltd., Berks, England). Thereafter, the tissues were cut into the cross-sectional
specimens by a microtome (Ultracuts, Leica Microsystems, Wetzlar,
Germany). The tissues were determined by the SEM (JSM-7800F, JEOL,
Tokyo, Japan) at an accelerating voltage of 5 kV.
3
Scanning Electron Microscopy Sample Preparation
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The specimens were fixed with 2 % paraformaldehyde and 2 % glutaraldehyde in 0.1 M sodium phosphate buffer (pH 7.4) and then stained with 1 % osmium tetroxide (Wako) in 0.1 M sodium phosphate buffer (pH 7.4). After the dehydration in a series of graded ethanol and the replacement by propylene oxide (Nacalai Tesque, Kyoto, Japan), the tissue was coated with a thin layer of platinum palladium using an ion sputter-coater (IB3, Eiko Corporation, Tokyo, Japan). Finally, images were obtained with a high-resolution scanning electron microscopy (S-4700, Hitachi High-Tech Corporation, Tokyo, Japan).
4
Scanning Electron Microscopy of Skin Cells
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Skin cells enlarged by collagen gels are purified with PBS and prepared in 4% glutaraldehyde (Wako, Osaka, Japan) in PBS for 24 hours scanning electron microscopy (SEM). After preparation, these cells are first immersed in 1% osmium tetroxide (Wako) for 1 hour. Following the dehydration of ethanol, the cells were at a critical point dried with liquid CO2, injected into SEM stubs with carbon tape and sprayed with gold. Samples were viewed and photographed with a scanning electron microscope (JSM-6701F, JEOL, and Tokyo, Japan). By measuring the area of the horizontal cell, the HDFs produced in the vessels are encased in crystal violet and photographed with a simple microscope (BX51, Olympus, and Tokyo, Japan). The location of the individual cell cells was mass-tested using the NIH Image software.
5
Ultrastructural Analysis of Intracranial Aneurysms
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At 5 days after the aneurysm induction, rats were killed as described previously. Dissected IA preparations were fixed in 4% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.4). Fixed specimens were then stained with 1% osmium tetroxide (Wako Pure Chemical Industries, Osaka, Japan) in 0.1 M sodium phosphate buffer (pH 7.4) for 1 h at room temperature. After dehydration in a series of graded ethanol solutions and replacement by propylene oxide (Nacalai Tesque), specimens were embedded in epoxy resin (Nacalai Tesque) overnight followed by the polymerization for 72 h. Ultrathin sections were prepared and images were obtained using an H‐7650 Transmission Electron Microscope (Hitachi, Tokyo, Japan).
6
Visualizing CHO Cell Morphology Before and After Cryopreservation
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The morphologies of CHO cells before and after ultra-quick freezing were visualized using scanning electron microscope (SEM). Briefly, the cell suspension was first fixed in a fixative containing 2.5% glutaraldehyde/0.1 M PBS (Wako) at room temperature for 30 min. After washing twice with 0.1 M PBS, the cells were post-fixed with 1% osmium tetroxide (Wako) at room temperature for 30 min. The cells were then washed twice with PBS, dehydrated through serial gradients of ethanol (10 min for each gradient), and finally dried using a critical point dryer. The cells were placed on carbon to obtain a thin layer and then coated with osmium using plasma CVD equipment. Each sample was observed under a SEM (JCM- 5700; JEOL).
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