Genetic recombination relies on a number of biochemical activities that must be present at the right time and place in order for two DNA molecules to be recombined properly. Recent advances in real-time fluorescence microscopy provide us with a glimpse of homologous recombination taking place in living cells. These approaches reveal that homologous recombination is highly choreographed in vivo with its spatio-temporal organization being dependent on both cell cycle phase and the nature of the initiating DNA lesion. In this chapter, we review the cell biology of homologous recombination in mitotic cells with the main focus on the yeast Saccharomyces cerevisiae but also drawing parallels to other eukaryotic organisms.