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Fixation Method Determines the Ultrastructure of the Nucleoid in Pseudomonas aeruginosa and Staphylococcus aureus
A E Grigor'eva1, E S Ryabova2, A V Tupitsyna2
1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia. feabelit@mail.ru.
Abstract:
The structure of the nucleoid observed after standard aldehyde fixation did not correspond to modern ideas about its spatial organization: DNA strands are not seen in Staphylococcus aureus and appear stuck together (2-15 nm) in Pseudomonas aeruginosa. The Ryter-Kellenberger (RK) fixation involving incubation in 1 % osmium tetroxide (pH 6.0) followed by 0.5 % uranyl acetate (pH 3.0) effectively preserves DNA strands (2 nm). The RK method also revealed additional details of the envelope structure in gram-positive and gram-negative bacteria. However, ribosomes, observed as spherical particles in aldehyde-fixed samples, were not visualized in the cytoplasm of S. aureus after RK fixation. The study provided a new information on the ultrastructure of bacteria, which is necessary for understanding the mechanisms of antibacterial compounds effects.
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The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
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