Appreciating the small things in life: STED microscopy in living cells

Abstract

Live-cell imaging has become indispensable in modern cell biology. However, many cellular processes can only be resolved by the means of super-resolution microscopy. Among the microscopy techniques that enable resolution beyond the diffraction limit of light, stimulated emission depletion (STED) microscopy is well suited for multicolour imaging of nanoscale dynamics. Here, we describe the development of live-cell STED imaging from proof-of-principle experiments into a versatile tool for multi-colour microscopic studies in living cells and even in living multicellular organisms. We summarise the advances in probes and hardware and discuss the current challenges and limitations that need to be addressed to realise the full potential of this technique.