Reversible Embedment Cytochemistry: A New Method for Ultrastructural Imaging and Affinity Labeling of Tissue Sections
| Autor: | G.G. Borisy, G. Gorbsky |
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| Rok vydání: | 1985 |
| Předmět: | |
| Zdroj: | Proceedings, annual meeting, Electron Microscopy Society of America. 43:434-437 |
| ISSN: | 2690-1315 0424-8201 |
| DOI: | 10.1017/s0424820100119004 |
| Popis: | Conventional epoxy and acrylic embedding procedures for electron microscopy involve infiltration into tissues of resin monomers that are subsequently polymerized and covalently crosslinked. During the polymerization and crosslinking reactions, tissue structure and antigenicity may be compromised by heat generation and by covalent modification of chemical subgroups within the tissue. In contrast, reversible embedment cytochemistry is a physical embedding procedure involving no chemical polymerization or covalent crosslinking reactions. In the procedure we have developed, fully polymerized polymethylmethacrylate was dissolved in dichloromethane and inflitrated into fixed and dehydrated tissue culture cells or tissues. The dichloromethane was allowed to evaporate at room temperature leaving the tissues embedded in plastic. Because polymethylmethacrylate is a hard, hydrophobic plastic, sections of from 30 nm to 1000 nm were reliably cut by conventional means using glass or diamond knives fitted with water troughs. Moreover, because polymethylmethacrylate is composed of linear polymers that are not covalently crosslinked, the plastic was easily reextracted from the sections by immersion in acetone. |
| Databáze: | OpenAIRE |
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