Cytochemical localization of polysaccharides in bacteria

Bernard, G. W.; Barkin, Michael E.

doi:10.1016/0003-9969(72)90121-5

Cited by 5 publications

(2 citation statements)

References 10 publications

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“…It has been reported that PTA has an affinity for polysaccharide components and stains the cell-surface coat in resin sections 17 . Polysaccharides are one of the major components of the bacterial cell envelope 18 , and the localization of polysaccharides in the bacterial cell wall has been demonstrated by transmission electron microscopy (TEM) of ultrathin sections stained with low pH PTA 19 . In this study, we confirmed the subcellular localization of tungsten by EDX in sections of bacteria stained with PTA before embedding and sectioning (Fig.…”

Section: Discussionmentioning

confidence: 99%

Detection of antimicrobial impact on gram-negative bacterial cell envelope based on single-cell imaging by scanning electron microscopy

Hisada

Matsumoto

Hirano

et al. 2023

Sci Rep

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Rapid determination of drug efficacy against bacterial pathogens is needed to detect potentially resistant bacteria and allow for more rational use of antimicrobials. As an indicator of the antimicrobial effect for rapid detection, we found changes in image brightness in antimicrobial-affected bacteria by scanning electron microscopy (SEM). The cell envelopes of unaffected bacteria were stained with phosphotungstic acid (PTA), whereas the entire cells of affected bacteria were stained. Since tungsten density increases backscattered electron intensity, brighter bacterial images indicate lethal damage. We propose a simplified method for determining antimicrobial efficacy by detecting damage that occurs immediately after drug administration using tabletop SEM. This method enabled the visualization of microscopic deformations while distinguishing bacterial-cell-envelope damage on gram-negative bacteria due to image-brightness change. Escherichia coli, Acinetobacter baumannii, Enterobacter cloacae, Klebsiella pneumoniae, and Pseudomonas aeruginosa were exposed to imipenem and colistin, which affect the cell envelope through different mechanisms. Classification of single-cell images based on brightness was quantified for approximately 500 bacteria per sample, and the bright images predominated within 5 to 60 min of antimicrobial treatment, depending on the species. Using intracellular PTA staining and characteristic deformations as indicators, it was possible to determine the efficacy of antimicrobials in causing bacterial-cell-envelope damage.

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Section: Discussionmentioning

confidence: 99%

Detection of antimicrobial impact on gram-negative bacterial cell envelope based on single-cell imaging by scanning electron microscopy

Hisada

Matsumoto

Hirano

et al. 2023

Sci Rep

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show abstract

“…This aligns with our observation of PTA distribution based on bacterial viability status, which affects the presence of these molecules on the bacterial surface. However, it is evident that this mechanism of PTA interaction depends on a number of chemical properties of the solution being used, including pH [42] . Some works describe the potential use of PTA in synthetic chemistry and its integration in nanoparticles, exploiting the unique chemical properties of the PTA molecule [43] , [44] .…”

Section: Discussionmentioning

confidence: 99%

Rapid microbial viability assay using scanning electron microscopy: a proof-of-concept using Phosphotungstic acid staining

Zmerli

Bellali

Haddad

et al. 2023

Computational and Structural Biotechnology Journal

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An ultrastructural study of the relationship of oral bacteria to the epithelium of healing tooth extraction wounds

McMillan

1975

Archives of Oral Biology

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Cytochemical localization of polysaccharides in bacteria

Cited by 5 publications

References 10 publications

Detection of antimicrobial impact on gram-negative bacterial cell envelope based on single-cell imaging by scanning electron microscopy

Detection of antimicrobial impact on gram-negative bacterial cell envelope based on single-cell imaging by scanning electron microscopy

Rapid microbial viability assay using scanning electron microscopy: a proof-of-concept using Phosphotungstic acid staining

An ultrastructural study of the relationship of oral bacteria to the epithelium of healing tooth extraction wounds

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