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Scanning probe microscopy for rheological analysis of biomolecular condensates

2025/11/08 by Aida Naghilou, Alireza Mashaghi · 1 voice · 1 citation
Biochemistry, Genetics and Molecular Biology · Physics and Astronomy · #Lipid Membrane Structure and Behavior #Force Microscopy Techniques and Applications #RNA Research and Splicing

paper · doi:10.1016/j.xpro.2025.104170

openalex publication_date 2025/11/08 · openalex created_date 2025/11/08 · openalex updated_date 2026/07/23

Abstract

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 • Protocol for measuring condensate rheology via scanning probe microscopy • Steps for condensate preparation, cantilever calibration, and rheology analysis • Data acquisition and analysis for viscoelasticity from force-distance curves Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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 condensates, 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.

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