Quantification of polysaccharides fixed to Gram stained slides using lactophenol cotton blue and digital image processing
نویسنده
چکیده
Dark blue rings and circles emerged when the non-specific polysaccharide stain lactophenol cotton blue was added to Gram stained slides. The dark blue staining is attributable to the presence of capsular polysaccharides and bacterial slime associated with clumps of Gram-negative bacteria. Since all bacterial cells are glycosylated and concentrate polysaccharides from the media, the majority of cells stain light blue. The contrast between dark and light staining is sufficient to enable a digital image processing thresholding technique to be quantitative with little background noise. Prior to the addition of lactophenol cotton blue, the Gram-stained slides appeared unremarkable, lacking ubiquitous clumps or stained polysaccharides. Adding lactophenol cotton blue to Gram stained slides is a quick and inexpensive way to screen cell cultures for bacterial slime, clumps and biofilms that are invisible using the Gram stain alone. The presence of cell clumping provides a possible explanation of the presence of persisters and paradoxical points observed in Virtual Colony Count antimicrobial assays, and suggests a phenotypic resistance mechanism to antimicrobial peptides involving capsular polysaccharides.
منابع مشابه
Quantification of polysaccharides fixed to Gram stained slides using lactophenol cotton blue and digital image
Dark blue rings and circles emerged when the non-specific polysaccharide stain lactophenol cotton blue was added to Gram stained slides. The dark blue staining is attributable to the presence of capsular polysaccharides and bacterial slime associated with clumps of Gram-negative bacteria. Since all bacterial cells are glycosylated and concentrate polysaccharides from the media, the majority of...
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I discovered indigo rings and circles in ATCC® 25922TM Escherichia coli cultures when I added the non-specific polysaccharide stain lactophenol cotton blue to Gram stained slides sampled from 96-well plates used to measure quantitative growth kinetics (QGK) in virtual colony count antimicrobial assays. I attribute the dark blue staining to the presence of capsular polysaccharides and bacterial ...
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