Flow Cytometry in Microbiology by David Lloyd (auth.), David Lloyd BSc, PhD, DSc (eds.)

By David Lloyd (auth.), David Lloyd BSc, PhD, DSc (eds.)

As but, circulation cytometry isn't used so generally in microbiology as in another disciplines. This quantity offers contributions movement cytometry to check a from examine microbiologists who use various set of difficulties. It illustrates the facility of the strategy, and should convince others of its usefulness. many of the con­ tributors amassed in Cardiff on 23 October 1991, at a gathering prepared for the Royal Microscopical Society via Dr. Richard Allman, however the content material in their chapters isn't really constrained through the discourse of that assembly, and for stability different specialists have been invited to write down for this ebook. stream Cytometry in Microbiology therefore represents the 1st selection of articles in particular dedicated to the purposes of a strategy which supplies loads to these investigating the microbial global. Cardiff, 1992 David Lloyd Contents record of individuals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix 1 circulation Cytometry: a method expecting Microbiologists David Lloyd . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 2 The actual and organic foundation for circulation Cytometry of Escherichia coli Erik Boye and Harald B. Steen . . . . . . . . . . . . . . . . . . . . . . eleven three movement Cytometric research of Heterogeneous Bacterial Populations Richard Allman, Richard Manchee and David Lloyd. . . . 27 four at the selection of the dimensions of Microbial Cells utilizing move Cytometry Hazel M. Davey, Chris L. Davey and Douglas B. Kell . . forty nine five makes use of of Membrane capability delicate Dyes with micro organism David Mason, Richard Allman and David Lloyd . . . . . . .

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6,um 3 , with a corresponding fluctuation in the forward scatter signal. However, the wide-angle scatter profile shows an almost inverse relationship to the forward scatter profile. In order to clarify this apparent anomaly we examined the samples used in the above experiment by electron microscopy. The results confirmed that the reduction in the wide-angle scatter signal for midexponential phase cells was caused by structural changes in the organisms (Fig. 3). The structural changes involved both internal and surface structures, though it was not possible to say what contribution each of these factors makes to the wide-angle scatter signal.

Cell type in any given instrument are proportional to cell size may not be completely correct. The processing of light scatter signals to obtain pulse-width information (time of flight) can yield more accurate estimates of cell size (Sharpless et al. 1975, 1977; Sharpless and Melamed 1976; Leary et al. 1979); however, such measurements are not feasible for bacteria, whose larger dimension is typically less than the width of the incident laser beam. Light scattered and collected orthogonally to the illumination source (socalled 90°, right-angle or wide-angle scatter) is also a complex parameter.

The way in which cells scatter light is rather complex and is dependent on their size, shape and internal structure. The relative contributions that these cell characteristics make to the amount of light that is scattered varies with the range of angles over which scattered light is collected, and in particular are functions of the relationship between the cell size and the wavelength of light (Steen and Lindmo 1985; Steen et al. 1989). Generally, the amount of forward scattering (small-angle scattering) increases rapidly with cell size and is affected to a much lesser extent by cell shape and refractive index, while the cell structure and shape become more important at larger angles (Paau et al.

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