Okajimas Folia Anat. Jpn., 56(1): 1-22, May 1979 New Embedding Method Employing GMA and Quetol 523 for Light and Electron Microscopic Observations of Semi-thin Sections
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A new method, employing water-miscible methacrylates, for light and electron microscopic observations of semi-thin sections was devised for ease in embedding, infiltration, sectioning and staining. The method used purified GMA (glycol methacrylate) and Quetol 523 (methoxy polyethylene glycol 400 methacrylate), and QCU-1 (2,2'-azobis isobutyronitrile paste) as catalyst. A mixture of these could be polymerized at 60°C for about 12hr or at 39°C for about 36 hr to produce a block with excellent cutting properties. Blocks of these resins were easy to section. Tissues were fixed in 2.5% glutaraldehyde with buffered phosphate at pH 7.4for 3 hr. Sections 0.3-2μm thick were cut with glass knives on a conventional ultramicrotome. They could be attached to slide glasses or grids by water flotation, without adhesive. They should be dried at room temperature. Staining with aqueous solutions of basic and acid dyes, without removing the embedding matrix, was sharp and brilliant as usual. These stained sections were observed with a light microscope. The same parts of such sections,0.3-0.5μm thick, could be examined with an accelerating voltage of 100 kV at low magnification (300-1,500 times) using a conventional electron microscope. Thus, photomicrographs and electron micrographs of the same part of a tissue sample could be exactly compared. The resolution of the electron microscope was so high that cytoplasmic components were easily identified in the cytoplasm. Osmium tetroxide vapor staining gave better contrast in the images of the specimens.
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