As part of the Imaging ONEWORLD series, the focus of these lectures is on microscopy and image analysis methods and how to apply these to your research. Almost all aspects of imaging such as sample preparation, labelling strategies, experimental workflows, ‘how-to’ image and analyse, as well as facilitating collaborations and inspiring new scientific ideas will be covered. Speakers will be available for questions and answers. The organisers, CRUK CI core facility staff, Gurdon Institute, MRC-LMB, MRC Cancer Unit and NPL will be able to continue the discussion and provide advice on your imaging projects.

Scientific Organisers

Imaging how bacteria sense and repair DNA damage at the single-molecule level

The accurate detection and repair of DNA damage is crucial for genome stability in all organisms. Despite extensive characterization of DNA repair pathways using genetics and biochemical assays, it remains unclear how repair proteins perform their function within the cellular environment. I will present our developments of microfluidic imaging techniques to investigate DNA repair and mutagenesis in individual living bacterial cells. By combining super-resolution localisation microscopy and single-molecule tracking, we were able to directly follow the movement and activity of DNA repair enzymes and regulatory proteins inside cells. This approach provided new insight into how DNA repair proteins identify lesion sites within the crowded cellular environment.

A key advantage of single-cell and single-molecule techniques is their ability to resolve biological heterogeneity and dynamic behaviour without ensemble averaging. Using these approaches, we found that stochasticity, or "noise", can play important roles in the function of the DNA repair system. By modulating mutation rates, noise in DNA repair can create a cell subpopulation that acts as a source of genetic diversity during adaptation to stress conditions.