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Abstract
Evidence is now mounting that liquid-liquid phase separation (LLPS) underlies the
formation of membraneless compartments in cells. This realization has motivated major
efforts to delineate the function of such biomolecular condensates in normal cells
and their roles in contexts ranging from development to age-related disease. There
is great interest in understanding the underlying biophysical principles and the specific
properties of biological condensates with the goal of bringing insights into a wide
range of biological processes and systems. The explosion of physiological and pathological
contexts involving LLPS requires clear standards for their study. Here, we propose
guidelines for rigorous experimental characterization of LLPS processes in vitro and
in cells, discuss the caveats of common experimental approaches, and point out experimental
and theoretical gaps in the field.