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. 2012 Jun 28;6(12):2178–2187. doi: 10.1038/ismej.2012.66

Figure 6.

Figure 6

Epifluorescence microscopy images of RCB encoding the dsrAB gene from sulfidic enrichment culture. Dual color hybridization of the 16S rRNA of RCB stained in green (Alexa 488 tyramide) and the dsrA targeting polynucleotide probe stained in red (Alexa 594 tyramide). Overlay images demonstrate that the signal for dsrA (red, a3 and a4) colocalizes with the 16S rRNA FISH signal for RCB (green, a4). Other bacteria in the enrichment (Epsilon- and Gammaproteobacteria) are stained with 4′,6-diamidino-2-phenylindole and are not fluorescent (a1). In addition, no fluorescent signals were obtained from hybridization of the NonPoly350 probe (b3) to the enrichment culture, which indicates the absence of nonspecific binding (b1–b4). a1, b1: DNA stains (4′,6-diamidino-2-phenylindole). a2: FISH signals of probe ROS537 targeting 16S rRNA (RCB specific). b2: FISH signals of probe GAM42a targeting 16S rRNA (Gammaproteobacteria). a3: FISH signals of dsrA gene-targeted probe. b3: negative control probe NonPoly350. a4: overlay images of 16S rRNA-FISH and dsrA-FISH. b4: overlay images of 16S rRNA-FISH and negative control probe NonPoly350. Scale bar corresponds to 5 μm.