Antimicrobial resistance (AMR) is one of today’s major global public health challenges. Recently, there has been renewed investment in the discovery of novel antimicrobials to urgently address the growing number of drug-resistant infections. This case study describes how Professor Gordon Dougan and his team at the Cambridge Biomedical Research Centre are using high-content analysis to phenotype individual bacteria within a population to investigate adaptive mechanisms of antimicrobial resistance, and to screen for novel alternatives to existing antimicrobials.