Imaging-Based Quantitative Assessment of Biomolecular Condensates in vitro and in Cells
The formation of biomolecular condensates contributes to intracellular compartmentalization, and plays an important role in many cellular processes. The characterization of condensates is however challenging, requiring advanced biophysical or biochemical methods that are often less suitable for in vivo studies. A particular need for easily accessible yet thorough methods that enable the characterization of condensates across different experimental systems thus remains. To address this, we present PhaseMetrics, a semi-automated FIJI-based image analysis pipeline tailored for quantifying particle properties from microscopy data. Tested using the FG-domain of yeast nucleoporin Nup100, PhaseMetrics accurately assesses particle properties across diverse experimental setups, including in vitro, Xenopus egg extracts, and cellular systems. It reliably detects changes induced by various conditions such as the presence of polyethylene glycol, 1,6-hexanediol, a salt gradient, and the molecular ch