. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. ORAL M1CROBIAL COMMUNITIES 203. Figure 2. Confocal micrographs of oral biofilms examined with fluorescence in xiin hybridization. (A) Mixed-species biofilm containing Streptococcus gonJonii DL1, Actinomyces naeslundii PKl°(a), and Fiisohnc- terium inicleatiim PKI594(f) grown on saliva in a flowed] for 4 h. The biofilm was stained with an FITC-labeled oligonucleotide probe (green) targeted to 5. gordonii and counterstained with the nucleic acid stain Syto 59 (red). Colocalized stains are yellow. Scale. 25 (B, C) Monospe
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. ORAL M1CROBIAL COMMUNITIES 203. Figure 2. Confocal micrographs of oral biofilms examined with fluorescence in xiin hybridization. (A) Mixed-species biofilm containing Streptococcus gonJonii DL1, Actinomyces naeslundii PKl°(a), and Fiisohnc- terium inicleatiim PKI594(f) grown on saliva in a flowed] for 4 h. The biofilm was stained with an FITC-labeled oligonucleotide probe (green) targeted to 5. gordonii and counterstained with the nucleic acid stain Syto 59 (red). Colocalized stains are yellow. Scale. 25 (B, C) Monospecies biofilms inoculated with S. f-urdiiii DLI and grown on enamel chips for 4 h. Biofilms were stained with an oligonucleotide probe specific for S. gordonii and the nucleic acid stain DAPI. Scale, 10 /j,m. (B) High-magnification image of enamel chip showing total number of cells visualized by DAPI stain. (C) Same location on enamel chip as in B. but only detecting cells labeled with oligonucleotide probe. with the growing biofilm. thus enabling bacterial species to be located without biofilm disruption (Fig. 2). Flowcells were consecutively inoculated with cultures of S. gordonii DLI, A. nticslundii PK19 (each an early colonizer), and Fusobacterium nucleunini PK1594 (late colonizer). After 4 h of saliva flow, biofilms were probed with a fluorescently labeled oligonucleotide designed to target streptococci (5'- GCTGCCTCCCGTAGGAGT-3'; JF20) as well as with a general nucleic acid stain to detect all cells (Fig. 2A). Based on distinctive morphologies of S. gordonii (coccus shaped). A. iKicsInndii (rod shaped), and F. nuclecituin (slender rods with tapered ends), all cell types could be visualized within the biofilm by staining with the nucleic acid marker Syto 59 (Molecular Probes, Eugene, OR). However, labeling by the fluorescent oligonucleotide probe was visible only in S. gordonii cells; the other two organisms did not bind the streptococcal probe (JF20) (Fig. 2A). RNA levels
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
