Scanning probe microscopy for rheological analysis of biomolecular condensates

Biomolecular condensates are membrane-less phase-separated assemblies that play key roles in cellular organization. Here, we present a protocol to characterize the full range of rheological properties of condensate droplets, from liquid to solid states, using scanning probe microscopy. We describe steps for preparing condensates, cantilever calibration, and condensate measurement. We then detail analysis procedures required to derive the complex shear modulus. For complete details on the use and execution of this protocol, please refer to Naghilou et al.¹.