A NEW TYPE OF BACTERIAL CELL WALL STRUCTURE REVEALED BY REPLICA TECHNIQUE.
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A new epoxy embedding mixture has been developed utilizing Maraglas 655 and Cardolite NC-513 with benzyldimethylamine (BDMA) as a curing agent. This epoxy mixture permits cellular preservation comparable to that obtained with Epon 812, ease of preparation of tissues, a wide range of miscibility, low viscosity, and, most important, ease of sectioning on a Porter-Blum microtome. In contrast to Epon-812-embedded tissues, Maraglas-Cardolite-embedded tissues can be sectioned in large dimensions with ease and consistent results without "chatter." No background granularity is detectable with high magnification study of Maraglas-Cardolite-embedded tissues. This epoxy is readily stained with lead hydroxide and is relatively stable in the electron beam.
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Bacillus polymyxa sporangia and spores were prepared for examination in the electron microscope by methods whose critical features were apparently: judicious use of vacuum, to encourage complete penetration of the embedding medium; the use of epoxy resins as embedding media; and cutting of the thin sections with a diamond knife. Electron micrographs of material prepared in this manner exhibit undeformed sporangial sections. Some of the structures revealed have been shown before, though perhaps less distinctly; other structures are revealed here for the first time. While this single study does not pretend to elucidate all the complexities of sporulation in bacteria, these and similar images should make this possible, and some mention of the preparatory techniques that lead to them seems advisable at this time.
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