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Condensate Growth Analysis Platform for Proteins Using Ultra-Widefield Dark-Field Microscopy and Image Analysis.

Kiyoto Kamagata^1,2,3, Ren Fujita², Eriko Mano¹

¹Faculty of Engineering, Gifu University, Yanagido 1-1, Gifu 501-1193, Japan.

The Journal of Physical Chemistry. B

|June 4, 2026

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Researchers developed a label-free microscopy platform to track biomolecular condensate growth. They found that condensates primarily grow via fusion, not Ostwald ripening, offering new insights into cellular organization.

Area of Science:

Cell Biology
Biophysics
Biochemistry

Background:

Biomolecular condensates are membraneless organelles crucial for cellular function.
They form via liquid-liquid phase separation, regulating biochemical reactions.
Understanding condensate formation and growth mechanisms is vital.

Purpose of the Study:

To develop a label-free platform for analyzing biomolecular condensate growth mechanisms.
To quantify the contributions of diffusion-limited growth, fusion, and Ostwald ripening.
To investigate the growth dynamics of HP1α and FUS condensates.

Main Methods:

Developed an ultra-widefield dark-field microscopy platform.
Enabled label-free, long-term (30 min), high-time-resolution (0.1 s) imaging over a 5.8 mm² area.

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Analyzed growth mechanisms for ~10,000 condensates in HP1α and FUS.

Main Results:

The platform successfully characterized condensate formation and growth without labeling.
HP1α and FUS condensates form via diffusion-limited growth.
Condensates predominantly grow through fusion, with minimal Ostwald ripening.

Conclusions:

The developed platform is effective for analyzing various biomolecular condensates in vitro.
Fusion is a primary growth mechanism for HP1α and FUS condensates.
Findings challenge previous assumptions about condensate growth dynamics.