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J Diabetes Res. 2014;2014:153586. doi: 10.1155/2014/153586. Epub 2014 Apr 15.

The visualization of biofilms in chronic diabetic foot wounds using routine diagnostic microscopy methods.

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  • 1Manchester Pharmacy School, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.
  • 2Department of Medicine Manchester Royal Infirmary, The University of Manchester, Oxford Road, Manchester M13 9PT, UK.
  • 3ConvaTec Ltd., Deeside CH5 2NU, UK.


Diabetic foot wounds are commonly colonised by taxonomically diverse microbial communities and may additionally be infected with specific pathogens. Since biofilms are demonstrably less susceptible to antimicrobial agents than are planktonic bacteria, and may be present in chronic wounds, there is increasing interest in their aetiological role. In the current investigation, the presence of structured microbial assemblages in chronic diabetic foot wounds is demonstrated using several visualization methods. Debridement samples, collected from the foot wounds of diabetic patients, were histologically sectioned and examined using bright-field, fluorescence, and environmental scanning electron microscopy and assessed by quantitative differential viable counting. All samples (n = 26) harboured bioburdens in excess of 5 log₁₀ CFU/g. Microcolonies were identified in 4/4 samples by all three microscopy methods, although bright-field and fluorescence microscopy were more effective at highlighting putative biofilm morphology than ESEM. Results in this pilot study indicate that bacterial microcolonies and putative biofilm matrix can be visualized in chronic wounds using fluorescence microscopy and ESEM, but also using the simple Gram stain.

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