Techniques for the detection and analysis of LLPS and MLOs

Liquid–liquid phase separation (LLPS) leading to the formation of various biomolecular condensates including membrane-less organelles (MLOs) represents the common mechanical principle of the intracellular space organization. MLOs can quickly assemble and disassemble under the influence of weak stimuli and act as important players of the cellular spatiotemporal organization. Distortion of the MLOs biogenesis or defilement of their properties leads to the development of various pathologies. Understanding of the basic biophysical principles and specific properties of MLOs can generate important insights into a wide range of biological processes. It also can help elucidating the causative relationships between the aberrant LLPS and the development of pathological conditions. All this requires clear standards for the analysis of LLPS and MLOs. This chapter provides a brief overview of the methods that are currently used to study assembly/disassembly of MLOs, characterize their structure and material properties, identify scaffold and client proteins, and analyze their functions.