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Isolation and visualization of viable mycobacteria in the RASC.

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posted on 2016-01-28, 12:41 authored by Robin Wood, Carl Morrow, Clifton E. Barry III, Wayne A. Bryden, Charles J. Call, Anthony J. Hickey, Charles E. Rodes, Thomas J. Scriba, Jonathan Blackburn, Chacha Issarow, Nicola Mulder, Jeremy Woodward, Atica Moosa, Vinayak Singh, Valerie Mizrahi, Digby F. Warner

(A) M. smegmatis::gfp growth on solid 7H10 agar plates from the Six-Stage Viable Andersen Cascade Impactor after wet release of 200 μl diluted culture into the RASC (30 000, 3000 and 300 colony forming units—CFU). The columns indicate the particle sizes captured on each plate across the 6 stages of the impactor, and the rows indicate the estimated total number of CFU passing through the impactor. Each release was repeated three times and the mean and SD for each plate are presented below the typical growth pattern distribution seen in the particle release. In all the releases the sampling was run for 5 minutes at 28 l/min resulting in the potential total capture of 3000, 300 and 30 CFU respectively. (B) SEM (left) and fluorescent microscopy (right) of M. smegmatis::gfp isolated on a PM10 impactor following experimental release.

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