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    Quantification of polysaccharides fixed to Gram stained slides using lactophenol cotton blue and digital image processing

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    Author
    Ericksen, B.
    Date
    2017
    Journal
    F1000Research
    Publisher
    Faculty of 1000 Ltd
    Type
    Article
    
    Metadata
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    See at
    https://doi.org/10.12688/f1000research.5779.1
    Abstract
    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. Copyright 2017 Ericksen B.
    Keyword
    antimicrobials
    Biofilms
    Escherichia coli
    Environment
    Identifier to cite or link to this item
    https://www.scopus.com/inward/record.uri?eid=2-s2.0-85037707881&doi=10.12688%2ff1000research.5779.1&partnerID=40&md5=64354586c37d11186e4b79361d16511f; http://hdl.handle.net/10713/10915
    ae974a485f413a2113503eed53cd6c53
    10.12688/f1000research.5779.1
    Scopus Count
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    UMB Open Access Articles 2017

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