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Updated: Aug 13, 2026
09:55
Force-Clamp Rheometry for Characterizing Protein-based Hydrogels
Published on: August 21, 2018
Stabilizing Protein and Protein-RNA Condensates in Low-Melting Agarose Hydrogels for Quantitative Biophysical
Nina C Kathe1, Leonidas Emmanouilidis2, Mihajlo Novakovic2
1Institute of Biochemistry, ETH Zürich; nina.kathe@bc.biol.ethz.ch.
Journal of Visualized Experiments : Jove
|August 10, 2026
Summary
Stabilizing biomolecular condensates in a hydrogel prevents variability, enabling precise, long-term biophysical measurements for cell biology research.
Area of Science:
- Cell Biology
- Biophysics
- Biochemistry
Background:
- Biomolecular condensates are crucial cellular structures formed by liquid-liquid phase separation.
- In vitro models of these condensates face challenges like coarsening and spreading, limiting quantitative analysis.
- Variability in measurements hinders a deep understanding of condensate function.
Purpose of the Study:
- To develop a method for stabilizing in vitro biomolecular condensates.
- To enable long-term and repeated quantitative measurements of condensate properties.
- To support advanced biophysical characterization of cellular condensates.
Main Methods:
- Embedding phase-separated biomolecular condensates in a low-melting agarose (LMA) hydrogel.
- Preparing LMA hydrogels at varying concentrations (0.3-1.0% w/v).
- Developing a step-by-step protocol for LMA stock preparation, droplet formation, and embedding.
Main Results:
- LMA hydrogels effectively stabilize biomolecular condensate droplets in three dimensions.
- Key droplet properties, such as molecular diffusion, are preserved post-embedding.
- The LMA embedding method is compatible with various advanced imaging and spectroscopy techniques.
Conclusions:
- Hydrogel embedding provides a robust solution to stabilize biomolecular condensates for quantitative studies.
- This technique enhances the reliability of biophysical measurements, advancing condensate research.
- The method facilitates deeper insights into the roles of condensates in health and disease.
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