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Flow-programmable and reversible surface-induced LLPS in nanofluidic channels.

Ryoichi Ohta1, Zhixin Zhao1, Xuan Yan1

  • 1Waseda University, 2-7 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan. ohta.ryo@aoni.waseda.jp.

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Researchers engineered a new method for precise control over liquid-liquid phase separation (LLPS) condensates on surfaces. This flow-programmable nanofluidic approach overcomes limitations of traditional surface functionalization for biomolecule concentration.

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Area of Science:

  • Biomolecular engineering
  • Nanofluidics
  • Cell biology

Background:

  • Cells utilize liquid-liquid phase separation (LLPS) to form membrane-less organelles for biomolecule concentration.
  • Mimicking cellular LLPS in engineered systems offers advantages over static surface functionalization but faces challenges in controlling condensate formation.

Purpose of the Study:

  • To develop a method for deterministic and instantaneous induction and manipulation of surface-mediated LLPS condensates.
  • To overcome the stochastic nature and limitations of bulk mixing for LLPS applications on a chip.

Main Methods:

  • Utilized nanochannels to leverage high surface area-to-volume ratios for surface-mediated LLPS.
  • Developed "flow-programmable nanofluidic surface LLPS" for controlled condensate formation and manipulation.
  • Investigated diffusion-limited transport physics governing condensate thickness (δ ∝ Q^0.3).

Main Results:

  • Achieved millisecond-scale equilibration and precise condensate thickness programming.
  • Demonstrated flow-tunable manipulation of condensates, including nm-scale thickness control and reversible hydrodynamic peel-off.
  • Showcased the potential for high-capacity functional molecule immobilization within 3D condensed phases.

Conclusions:

  • Flow-programmable nanofluidic surface LLPS enables deterministic control over biomolecular condensates.
  • This method overcomes limitations of conventional surface functionalization for applications requiring high molecule capacity and dynamic control.
  • The technology holds promise for advanced applications like substrate channeling and dynamic reactor operation.