[Ultrastructure of B. pertussis using cryoultramicrotomy].
The authors pointed to a possibility of using the method of cryoultramicrotomy in studying the ultrastructure of Gram negative B. pertussis bacteria under the following conditions: a) fixationwith a 5% glutaraldehyde on cacodylate buffer (pH 7.5) for 30 min; b) replacement with a 30% dimethylsulfoxide in distilled water with a subsequent embedding of the material into tissue-tek; c) freezing in fluid nitrogen for 5 min; d) cutting at a temperature of -- 90% degrees C and at the temperature of glass knife of -- 40 degrees C, the knife angle of alpha = 45 degrees, beta = 6 degrees, and the cutting velocity of 5 mm/sec; e) placing of sections on copper grates covered with a 0.1% moulding membrane; d) contrasting with a 2% phosphotungstic acid queous solution (pH 7.5) for 5 sec and with a 3% uranil acetate on water for 5 min. Use of the mentioned method pemitted to detect structures localized in B. pertussis cytoplasm and also to trace the dynamics of their formation.
D001886:Bordetella pertussis / Q000648:ultrastructure*; D008867:Microtomy / Q000379:methods*
Izuchenie ul'trastruktury B. pertussis metodom krioul'tramikrotomii