Mature RAW-derived osteoclasts were fixed in 2% glutaraldehyde in a 0.1 M sodium cacodylate buffer at pH 7.2 for 2 h at room temperature. Osteoclasts were postfixed in 1% osmium tetroxide and 1.25% potassium ferrocyanide in sodium cacodylate buffer at room temperature for 45 min and stained for 30 min with 4% uranyl acetate in water. Dehydration was carried out by incubating in a graded series of ethanol ranging from 70 to 100%. To embed the cells, samples were infiltrated with a 1:1 mixture of SPI-812 epoxy (SPI Supplies, West Chester, PA) and ethanol for 45 min, followed by two incubations with 100% epoxy for 120 min and overnight subsequently. After hardening of the resin, ultrathin sections (80 nm) were cut and collected onto copper grids prior to being poststained with uranyl acetate and lead citrate. Sections were imaged using a Hitachi H7500 transmission electron microscope with images being captured with an Olympus SIS Megaview II digital camera.
Sis megaview 2
Manufactured by Olympus
The SIS Megaview II is a digital camera system designed for microscopy applications. It features a high-resolution CMOS sensor and advanced image processing capabilities to capture detailed micrographs. The SIS Megaview II is capable of providing real-time image acquisition and display for a wide range of microscopy techniques.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sis megaview 2
1
Ultrastructural Analysis of Mature Osteoclasts
2
Visualizing Amyloid-Beta Aggregation
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Samples (5 μL) taken at the 12 h time point from aggregation reactions containing Aβ1–42 alone, or Aβ1–42 mixed with pCLU or rCLU-ST, were applied to Formvar-coated copper grids. The samples were stained with 2% (w/v) uranyl acetate, and imaged using a FEI Tecnai T12 transmission electron microscope (Cryogenic Electron Microscopy Facility, Molecular Horizons, University of Wollongong). Images were analysed using the SIS Megaview II Image Capture system (Olympus).
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