We aim to understand how different bacteria can grow across a large temperature range from 0°C to 100°C. We focus on how temperature affects the cytoplasmic state in bacteria and what the consequences to cellular processes are. To study this, we use super-resolution microscopy and single molecule tracking to record the movement of individual molecules, such as ribosome, inside individual live cells. Single molecule dynamics report on the molecular interactions in real-time, acting as a probe for changes in the cellular environment. The obtained knowledge will help us to understand microorganisms’ adaptation to different temperatures and the limits of cellular life. The research will also contribute to the development of robust synthetic biology applications and novel assay solutions.