Two novel experimental approaches have been developed to investigate the attachment of microorganisms to sulphide minerals as a function of the mineral and microbial phase in a joint project with BHP Billiton, conceptualised in 2005 and initiated in 2006. In the first approach, selective attachment of pure cultures to minerals was studied in the “particle coated column reactor” using A. ferrooxidans and L. ferriphilum. The saturated reactor containing glass beads coated with fine mineral concentrates provided a quantifiable surface area of mineral concentrate and maintained good fluid flow. Results are reported for chalcopyrite and pyrite concentrates, a low grade chalcopyrite ore and quartzite. The latter, representing typical gangue material, is used as a control. A. ferrooxidans displayed greater attachment to pyrite, and selective attachment to sulfide minerals over quartz. Similar attachment behaviour trends resulted for Leptospirillum spp. In the second approach, a novel technique was developed to investigate microbial ecology of microbe-mineral attachment, site and mineral specific associations of microorganisms and spatial organisation of microbial communities present. Qualitative assessment and visualisation of microorganisms associated with the mineral surface and subsequent biofilm development was shown in the biofilm reactor, using microscopy techniques and fluorochromes. FISH analyses of A. ferrooxidans and L. ferriphilum on massive chalcopyrite sections are presented. The consequence of the observed attachment on heap bioleach performance is discussed.