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FIGURE 6.4 CLSM micrographs of a bacterial microcolony stained with lectins (A) Cicer arietinum-Alexa-568; (B)Solanum tuberosum-FITC; (C) Tetragonolobuspurpureas-CY5; and (D) the overlay image of all three channels showing the layers and differential lectin binding.

FIGURE 6.5 (Color Figure 6.5 appears following page 236.) Images of the combined FLBA-FISH approach showing (A) staining of an Elbe River floc where the gene probe EUB-CY3 and lectin Canavalia ensiformis-FITC were applied; and (B) the binding of the lectin Cicer arietinum-Alexa-568 and the lectin Arachis hypogaea-CY5 with localization of beta-proteobacterial cells using the probe Bet42a.

image analyses of confocal image stacks, to quantitatively evaluate the binding of different lectins spatially and with time. Neu et al.18 were able to detect clear statistically significant effects of nutrient treatments and time on the EPS composition of river biofilms using CLSM and FLBA. Figure 6.6 shows a typical data set with variation in lectin binding in response to the addition of nutrients during biofilm development. There were however, effects of the fluor, the matrix, and the lectin on the apparent specificity of lectin binding and limitations on the interpretation of the nature of binding site of the lectin. Recent comparative STXM-CLSM studies of biofilms demonstrated significant agreement between the probe target dependent

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