Infection of Vero cells with Coxiella burnetii phase II: relative intracellular bacterial load and distribution estimated by confocal laser scanning microscopy and morphometry
Ds. Zamboni et al., Infection of Vero cells with Coxiella burnetii phase II: relative intracellular bacterial load and distribution estimated by confocal laser scanning microscopy and morphometry, J MICROB M, 43(3), 2001, pp. 223-232
Coxiella burnetii, the agent of Q fever in man and of coxiellosis in other
species, is an intracellular pathogen not yet grown axenically. Confocal la
ser fluorescence microscopy and morphometry were used to measure relative C
. burnetii phase II loads and their intracellular distribution in aldehyde
fixed and DAPI stained Vero cell monolayers. The fluorescence of single hor
izontal optical sections provided useful information on relative loads of b
acteria in cells and vacuoles. The relative density of the bacteria in the
vacuoles was inferred from ratios of fluorescence to vacuolar section areas
. Relative bacterial loads, bacterial densities and section areas of large
vacuoles increased exponentially between days 2 and 4 of the infection of g
amma -irradiated host cells, stabilized between days 4 and 6, and decreased
thereafter. Estimated minimum doubling times were higher for the overall c
omplement of the intracellular organisms (about 12 h) than for bacteria tha
t were confined to larger vacuoles (about 10 h), (C) 2001 Elsevier Science
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