Osmium tetroxide solution

Manufactured by Agar Scientific

Sourced in United Kingdom

Osmium tetroxide solution is a colorless, volatile liquid that is used as a fixative and staining agent in various laboratory applications. It is a powerful oxidizing agent and must be handled with caution.

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Lab products found in correlation

Nrecon v1, by Bruker (2 mentions) Skyscan 1172, by Bruker (2 mentions) Sodium cacodylate buffer, by Merck Group (2 mentions) Axiophot 2 microscope, by Zeiss (2 mentions) Copper grid, by Merck Group (2 mentions) Glutaraldehyde solution, by Merck Group (2 mentions) Uranyl acetate, by Agar Scientific (2 mentions) Lead citrate, by Agar Scientific (2 mentions) Tecnai 10, by Philips (1 mentions) Mega view 2, by Philips (1 mentions) Diamond knife, by Electron Microscopy Sciences (1 mentions) Ultramicrotome, by Reichert Technologies (1 mentions)

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Osmium tetroxide (71 citations)

6 protocols using osmium tetroxide solution

Decalcification and Osmium Staining for μCT

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Tibiae were isolated and, after removal of external soft tissue, fixed in formalin at 4°C. Fixed tibiae were decalcified in 14% EDTA for 14 days and then washed in Sorensen’s Phosphate buffer (81 mM KH₂PO₄, 19 mM Na₂HPO₄ ⋅ 7H₂O, pH 7.4). Decalcified tibiae were stored in Sorensen’s Phosphate buffer at 4°C until ready to be stained with osmium tetroxide. To do so, osmium tetroxide solution (2% w/v; Agar Scientific, UK) was diluted 1:1 in Sorensen’s Phosphate buffer. Tibiae were then stained in this 1% osmium tetroxide solution for 48 h at room temperature, then washed, and stored in Sorensen’s Phosphate buffer at 4°C prior to micro computed tomography (μCT) analysis.

Sulston R.J., Learman B.S., Zhang B., Scheller E.L., Parlee S.D., Simon B.R., Mori H., Bree A.J., Wallace R.J., Krishnan V., MacDougald O.A, & Cawthorn W.P. (2016). Increased Circulating Adiponectin in Response to Thiazolidinediones: Investigating the Role of Bone Marrow Adipose Tissue. Frontiers in Endocrinology, 7, 128.

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Tibial Microarchitecture Analysis by μCT

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Following μCT analysis at 12 weeks, bones were removed from 1% agarose and washed in Sorensen's phosphate buffer (81mM KH₂PO₄, 19mM Na₂HPO₄ · 7H₂O, pH 7.4) prior to decalcification. Tibia were decalcified in 14% EDTA for 2 weeks at 4°C, washed in Sorensen's phosphate buffer and stained with 1% osmium tetroxide solution (1% w/v; diluted 1:1 in Sorensen's phosphate buffer; Agar Scientific, Stansted, Essex, UK) for 48 hours at room temperature, washed, and stored in Sorensen's phosphate buffer at 4°C. Stained tibias were embedded in 1% agarose and scanned using a Skyscan 1172 desktop micro‐CT (Bruker) as previously described.⁽³¹, ³²⁾ The data were reconstructed using Skyscan software NRecon v1.6.9.4 (Bruker). Volumetric analysis was performed using CTAn v1.13.5.1 (Bruker).

Dillon S., Suchacki K., Hsu S., Stephen L.A., Wang R., Cawthorn W.P., Stewart A.J., Nudelman F., Morton N.M, & Farquharson C. (2020). Ablation of Enpp6 Results in Transient Bone Hypomineralization. JBMR Plus, 5(2), e10439.

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Micro-CT Analysis of Decalcified Tibias

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Formalin‐fixed, decalcified tibias were washed in Sorensen's phosphate buffer (81 mM KH2PO4, 19 mM Na2HPO4 · 7H2O, pH 7.4) and stained with 1% osmium tetroxide solution (2% w/v; Agar Scientific, Stansted, UK; diluted 1:1 in Sorensen's phosphate buffer) for 48 hours at room temperature, washed, and stored in Sorensen's phosphate buffer at 4°C. Stained tibias were arranged in parallel in 1% agarose in a 30‐mL universal tube and mounted in a SkyScan 1172 desktop micro‐CT (Bruker, Kontich, Belgium). The samples were then scanned through 360° using a step of 0.40° between exposures. A voxel resolution of 12.05 μm was obtained in the scans using the following control settings: 54 kV source voltage, 185 μA source current with an exposure time of 885 ms. A 0.5‐mm aluminium filter and two‐frame averaging were used to optimize the scan. After scanning, the data were reconstructed using SkyScan software NRecon v1.6.9.4 (Bruker). The reconstruction thresholding window was optimized to encapsulate the target image. Volumetric analysis was performed using CT Analyzer v1.13.5.1 (Bruker).24 (link)

Morris E.V., Suchacki K.J., Hocking J., Cartwright R., Sowman A., Gamez B., Lea R., Drake M.T., Cawthorn W.P, & Edwards C.M. (2020). Myeloma Cells Down‐Regulate Adiponectin in Bone Marrow Adipocytes Via TNF‐Alpha. Journal of Bone and Mineral Research, 35(5), 942-955.

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Transmission Electron Microscopy Specimen Preparation

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For TEM investigations, specimens were fixed in 2.5% glutaraldehyde solution (Sigma-Aldrich Chemie GmbH, Germany) buffered with sodium cacodylate buffer (Sigma-Aldrich Chemie GmbH, Germany) at pH = 7.4 for two hours at 4 °C. The samples were postfixed for 1 h in 1% osmium tetroxide solution (Agar Scientific, Stansted, UK) at the same temperature and pH, dehydrated at increasing concentrations of ethanol (50, 70, 90, 96, 99.5%), in an acetone series and embedded in Epon-812 (Fluka Chemie AG, Buchs, Switzerland) according to standard methods. Semithin sections were stained with methylene blue-azur II to select the region of interest for the following procedures. The semithin sections were analysed using a Zeiss Axiophot 2 microscope (Zeiss, Germany). The ultrathin (80 nm) sections were cut on the Reichert Om U3 ultratome with a diamond knife (Diatome Ltd, Biel/Bienne, Switzerland). Sections were mounted on copper grids of mesh size 200 (Sigma-Aldrich Chemie GmbH, Germany) with Perfect Loop (Diatome Ltd., Biel/Bienne, Switzerland) and stained with uranyl acetate (Agar Scientific, Stansted, UK) and lead citrate (Agar Scientific, Stansted, UK) according to standard methods. For TEM, a Philips Tecnai-10 with camera Mega View II was used for viewing and photographing.

Aunapuu M., Kibur P., Järveots T, & Arend A. (2018). Changes in Morphology and Presence of Pinopodes in Endometrial Cells during the Luteal Phase in Women with Infertility Problems: A Pilot Study. Medicina, 54(5), 69.

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Transmission Electron Microscopy Specimen Preparation

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For TEM investigations, specimens were fixed in 2.5% glutaraldehyde solution (Sigma-Aldrich Chemie GmbH, Germany) buffered with sodium cacodylate buffer (Sigma-Aldrich Chemie GmbH, Germany) at pH = 7.4 for 2 h at 4 °C. The samples were postfixed for 1 h in 1% osmium tetroxide solution (Agar Scientific, Stansted, UK) at the same temperature and pH, dehydrated at increasing concentrations of ethanol (50%, 70%, 90%, 96%, and 99.5%), in an acetone series and embedded in Epon-812 (Fluka Chemie AG, Buchs, Switzerland) according to standard methods. Semithin sections were stained with methylene blue-azur II to select the region of interest for the following procedures. The semithin sections were analyzed using a Zeiss Axiophot 2 microscope (Zeiss, Germany). The ultrathin (80 nm) sections were cut on the Reichert Om U3 ultratome with a diamond knife (Diatome, Ltd., Biel/Bienne, Switzerland). Sections were mounted on copper grids of mesh size 200 (Sigma-Aldrich Chemie GmbH, Germany) with Perfect Loop (Diatome, Ltd., Biel/Bienne, Switzerland) and stained with uranyl acetate (Agar Scientific, Stansted, UK) and lead citrate (Agar Scientific, Stansted, UK) according to standard methods. For TEM, Tecnai G2 spirit 120 kV frozen TEM was used for viewing and photographing.

Lin Y., Li Y., Chen P., Zhang Y., Sun J., Sun X., Li J., Jin J., Xue J., Zheng J., Jiang X.C., Chen C., Li X., Wu Y., Zhao W., Liu J., Ye X., Zhang R., Gao J, & Zhang D. (2023). Exosome-Based Regimen Rescues Endometrial Fibrosis in Intrauterine Adhesions Via Targeting Clinical Fibrosis Biomarkers. Stem Cells Translational Medicine, 12(3), 154-168.

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Ultrastructural Analysis of Trabecular Bone

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Bone tissue was fixed in a buffered (0.1 M sodium cacodylate buffer, pH 7.4) 2.0% glutaraldehyde solution for 18 h at 4 °C and decalcified in EDTA. The specimens were cut into small pieces (1–3 mm3) early in the chemical fixation procedure. The tissue was washed (2 × 30 min) in buffer followed by secondary fixation in a 1% osmium tetroxide solution (Agar Scientific Ltd., UK) diluted in 0.1 M sodium cacodylate, pH 7.4. The tissue was dehydrated through a graded series of ethanols (70–100%) and placed in propylene oxide/Spurr’s resin (1:1) before vacuum infiltration in freshly prepared Spurr’s resin for 2 days and then a final resin change for polymerization at 60 °C for 18 h. An area encompassing trabecular bone was selected and ultrathin sections were cut on a Reichert-Jung ultramicrotome at 40–60 nm, contrasted using aqueous uranyl acetate for 10 min followed by lead citrate before viewing in the transmission electron microscope [28 (link)].
For scanning electron microscopy, bone tissue was fixed in buffered 2.0% glutaraldehyde solution (18 h) at 4 °C. Samples were examined after critical point drying and sputter coating with gold.

Barnes A.M., Ashok A., Makareeva E.N., Brusel M., Cabral W.A., Weis M., Moali C., Bettler E., Eyre D.R., Cassella J.P., Leikin S., Hulmes D.J., Kessler E, & Marini J.C. (2019). COL1A1 C-propeptide mutations cause ER mislocalization of procollagen and impair C-terminal procollagen processing. Biochimica et biophysica acta. Molecular basis of disease, 1865(9), 2210-2223.

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