High‐Resolution Microscopy for Structural Studies of Biological Systems in Cells

Abstract

Atomic resolution structures of proteins are essential for understanding the biological roles played by these molecules. However, in order to understand fully how the molecules function in living systems, other techniques must be applied that provide insights into how they interact in cells and tissues. No single technique can provide all the information needed, but a multi‐technique approach can be applied to provide information on different aspects of molecular structure and interaction in cells and tissues. Methods that can be applied include a range of fluorescence‐based microscopy techniques, including ‘super‐resolution’ microscopy, which provides subdiffraction limit images. When accompanied by other techniques such as single particle tracking, electron microscopy and molecular dynamics simulation, these methods can provide new insights into the functioning of complex biological processes. Key Concepts Linking structure and function requires methods that can provide structural information on molecules and molecular complexes in cells. ‘Super‐resolution’ optical methods that can image below the diffraction limit allow us to fill in the resolution gap between conventional optical and electron microscopy. Single‐molecule techniques allow us to investigate individual interactions without the requirement to synchronise cellular processes. Often no single technique can provide all the information needed so an approach combining different imaging methods is often required. Molecular dynamics simulations can help in the interpretation of in cellulo structural data.