Osmium tetraoxide

Manufactured by Ted Pella

Sourced in China

Osmium tetraoxide is a chemical compound with the formula OsO4. It is a colorless crystalline solid that sublimes at room temperature. Osmium tetraoxide is a strong oxidizing agent and is commonly used in various laboratory applications.

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Lab products found in correlation

Jem 1400 transmission electron microscope, by JEOL (1 mentions) Glutaraldehyde, by Ted Pella (1 mentions) Libra 120 eftem, by Zeiss (1 mentions)

3 protocols using osmium tetraoxide

Ultrastructural Analysis of Cells

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Cells (1×10⁶) were fixed in 2.5% glutaraldehyde (Ted Pella Inc., Redding, CA, USA) and 0.1 mol/l phosphate buffer (pH 7.4; Jinuo, Hangzhou, China) for 30 min at room temperature followed by fixation in 1% osmium tetraoxide (Ted Pella Inc.) and 0.1 mol/l phosphate buffer (pH 7.4) for 30 min. Cells were then subjected to graded ethanol dehydration with epoxy propane (Ted Pella Inc.), embedded in epoxy resin and segmented using a UC7 ultrastructural slicer (Leica Microsystems GmbH, Wetzlar, Germany). Sections (90-nm) were subsequently stained with 5% uranyl acetate (SPI-Chem, West Chester, PA, USA) and 5% lead citrate (SPI-Chem) prior to analysis using a JEM1400 transmission electron microscope (JEOL Ltd., Tokyo, Japan).

Liu J., Ren H., Liu B., Zhang Q., Li M, & Zhu R. (2016). Diosmetin inhibits cell proliferation and induces apoptosis by regulating autophagy via the mammalian target of rapamycin pathway in hepatocellular carcinoma HepG2 cells. Oncology Letters, 12(6), 4385-4392.

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Centrosome Amplification and Organization

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To study centrosome amplification and organization in higher magnification 14-3-3γ-knockdown and vector-control cells were visualized under transmission electron microscope. Synchronized cells in S-phase were fixed with 3% glutaraldehyde, washed with 0.1 M of sodium cacodylate and post fixed with 1% osmium tetra oxide (Tedpella). Cultures were dehydrated and processed. Grids were contrasted with alcoholic uranyl acetate for 1 minute and lead citrate for half a minute. The grids were observed under a Carl Zeiss LIBRA120 EFTEM transmission electron microscope, at an accelerating voltage of 120KV and at 25000X magnification. Images were captured using a Slow Scan CCD camera (TRS, Germany).

Mukhopadhyay A., Sehgal L., Bose A., Gulvady A., Senapati P., Thorat R., Basu S., Bhatt K., Hosing A.S., Balyan R., Borde L., Kundu T.K, & Dalal S.N. (2016). 14-3-3γ Prevents Centrosome Amplification and Neoplastic Progression. Scientific Reports, 6, 26580.

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Localization of Quantum Dots in HaCaT Cells

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HaCaT cells were co-cultured with the 10 nM of GSH-, DHLA-, CYS-, PEI-capped QDs for 24 h as described above. To investigate the location of the QDs in the HaCaT cells, the cells were prepared for TEM analysis. The cells were fixed with 2.5% glutaraldehyde in 0.1 M, pH 7.2 cacodylate buffer for 24 h at 4 °C and rinsed with cold 0.1 M pH 7.2 cacodylate buffer three times. The cells were then fixed with 1% osmium tetraoxide (Ted Pella, Inc.) for 1 h at 4°C and washed with cold distilled water three times. The cells were dehydrated using graded alcohol baths (25%, 50%, 75%, and 100%) and then infiltrated with and embedded in Spurr epoxy resin with overnight polymerization at 70 °C. After embedding, the samples were cut to 1−2 μm with a glass blade and finally sliced at 70 nm with a diamond knife and placed on copper grids. Nanoparticle localization was evaluated using a Hitachi 5100 TEM apparatus.

Zheng H., Mortensen L.J., Ravichandran S., Karen B, & DeLouise L.A. (2017). Effect of nanoparticle surface coating on cell toxicity and mitochondria uptake. Journal of biomedical nanotechnology, 13(2), 155-166.

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