The complex interactions between patients and the microbial species that reside in their wounds are not yet fully characterised. Investigations to date have dealt with either those organisms that cause infections or those that establish long-term colonisations. The objective of this open, prospective pilot study was to quantitatively and qualitatively assess the microbiological status of ‘locally infected’ venous leg ulcers. Three different sampling techniques were used to recover organisms from 20 venous leg ulcers that had failed to progress towards healing and comparisons made to explore the distribution of colonised flora within the wound. In total, 116 isolates were recovered (mean of 5·8 per ulcer) with highest recovery rates seen with swabs. Greatest agreement of colonisation and counts was found between swabs and absorbent polyvinyl acetate (PVA) foam disc, and also between PVA discs and biopsies. Lowest agreement was between the distribution of anaerobes and coryneforms in swabs and biopsies, suggesting uneven vertical distribution within ulcers. No justification for using routine biopsies in locally infected venous leg ulcers was found.