Abstract
This transmission electron microscopy (TEM) procedure was designed to examine a gram positive spore-forming bacillus in colony on various solid agar media with minimal artifact. Cellular morphology and organization of colonies embedded in Poly/Bed 812 resin (P/B) were studied. It is a modification of procedures used for undecalcified rat bone and Stomatococcus mucilaginosus.Cultures were fixed and processed at room temperature (RT) under a fume hood. Solutions were added with a Pasteur pipet and removed by gentle vacuum aspiration. Other equipment used is shown in Figure 3. Cultures were fixed for 17-18 h in 10-20 ml of RT 2% phosphate buffered glutaraldehyde (422 mosm/KgH2O) within 5 m after removal from the incubator. After 3 (30 m) changes in 0.15 M phosphate buffer (PB = 209-213 mosm/KgH2O, pH 7.39-7.41), colony cut-outs (CCO) were made with a scalpel.
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More From: Proceedings, annual meeting, Electron Microscopy Society of America
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