By Samuel Razin (Eds.)
Equipment in Mycoplasmology V1
summary: tools in Mycoplasmology V1
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Additional resources for Methods in Mycoplasmology. Mycoplasma Characterization
115, 367-386. Freundt, E. A. (1959). " Munksgaard, Copenhagen. Razin, S. (1978). The mycoplasmas. Microbiol. Rev. 42, 414-470. , and Cosenza, B. J. (1966). Growth phases of Mycoplasma in liquid media observed with phase-contrast microscope. J. Bacteriol. 91, 858-869. Subcommittee on the Taxonomy of Mollicutes (1979). Proposal of minimal standards for descriptions of new species of the class Mollicutes. Int. J. Syst. Bacteriol. 29, 172-180. B2 PHASE-CONTRAST MICROSCOPY W. Bredt INTRODUCTION As a result of the lack of a cell wall, the cellular shape of mycoplasmas is greatly distorted when the routine fixation and staining procedures used for bacteria are applied to mycoplasmas.
10. After 30 to 60 seconds, withdraw the drop of stain in the manner described in (8). 11. Allow to dry for a minute or two and examine in the electron microscope. Agar-Grown Mycoplasmas (Colonies or Lawns) 1. Place a grid, coated surface down, on the lawn or colony (or if monitoring for viruses, on a plaque). 2. Press very gently on the grid, and allow it to remain for 1 minute or longer. 3. Remove the grid, place on sticky tape on a slide as in the preceding step (5), and immediately apply a drop of 2% ammonium molybdate as in preceding step (9).
2. Rinse the samples in phosphate buffer for 2-24 hours. 3. 1 M phosphate buffer for 1-2 hours. 4. Rinse samples in phosphate buffer for 15-30 minutes. 5. Perform dehydration series as indicated: 35% 50% 70% 95% 95% 100% 100% EtOH EtOH EtOH EtOH EtOH EtOH EtOH 10 10 10 10 10 15 15 minutes minutes minutes minutes minutes minutes minutes 6. Perform infiltration as follows: Propylene oxide Propylene oxide 1:1 Propylene oxide:epoxy Epoxy 10 minutes 10 minutes Overnight 2 hours B7 Specialized Electron Microscopic Techniques 59 7.
Methods in Mycoplasmology. Mycoplasma Characterization by Samuel Razin (Eds.)