Abstract
Publisher Summary This chapter discusses the procedures for low-temperature embedding in Lowicryl K4M together with various protocols for the light and electron microscopic localization of cellular constituents as they have been used in laboratory. The development of the Lowicryl resins, particularly the hydrophilic Lowicryl K4M, for low-temperature embedding by Carlemalm er al. (1982) and their successful introduction for postembedding immunolabeling can be considered a major achievement. This low-temperature embedding technique has not only provided superior preservation of antigens, but has also resulted in improved preservation of fine-structural details of mildly aldehyde-fixed tissues and cells, and a drastically reduced background staining. In addition, it dramatically improved the detectability of glycoconjugates with lectins, monoclonal antibodies, and glycosyltransferases. The basic principles of sectioning resin-embedded materials apply to Lowicryl K4M. Lowicryl K4M blocks can be sectioned with glass or diamond knives. The angle of the pyramids should be in the range of 55˚-60˚. It is recommended to trim the final pyramids with glass knives or on a trimming machine. For sectioning, precautions have to be taken to prevent wetting of the pyramid because Lowicryl K4M is hydrophilic.
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