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Fig. 9. Whole-mount staining of control and dimB- mutant standing
slugs. dimB- and control (random integrant) strains were created
containing ecmAO:lacZ. After development to the standing slug stage, slugs
were stained with a blue ß-galactosidase chromogen. Quantitative analysis
of additional such images was performed to measure the areas occupied by the
prestalk and prespore zones. These measurements were taken from photographic
images, using the `magic wand' tool in `Adobe Photoshop' to select areas with
similar signal densities. This revealed more heterogeneity in the apparent
fraction of prestalk cells in the mutant but, on average, there was no major
difference from the control (control=22±4.4%, n=14;
dimB null=19±8.9%, n=15). The greater heterogeneity
of the dimB-null slugs probably results from their failure to migrate
away from their point of origin and from their tendency to break up into
fragments. For double staining analysis, dimB- and control (random
integrant) strains were created containing a pspA:glucoronidase reporter and
one of the three prestalk lacZ reporter fusions indicated at the left. The
strains were developed on water agar to the standing slug stage and the slugs
were fixed and stained sequentially for ß-glucoronidase (blue) and
ß-galactosidase (red).
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