Nakazawa Koyomi, Lévrier Antoine, Rudiuk Sergii, Yamada Ayako, Morel Mathieu, Baigl Damien
PASTEUR, Department of Chemistry, École Normale Supérieure, PSL University, Sorbonne Université, CNRS, 75005, Paris, France.
Angew Chem Int Ed Engl. 2025 Feb 10;64(7):e202419529. doi: 10.1002/anie.202419529. Epub 2025 Jan 7.
Giant unilamellar vesicles (GUVs) are widely used minimal cell models where essential biological features can be reproduced, isolated and studied. Although precise spatio-temporal distribution of membrane domains is a process of crucial importance in living cells, it is still highly challenging to generate anisotropic GUVs with domains at user-defined positions. Here we describe a novel and robust method to control the spatial position of lipid domains of liquid-ordered (Lo)/liquid-disordered (Ld) phase in giant unilamellar vesicles. Our strategy consists in confining Lo/Ld phase-separating GUVs in microfluidic channels to define free curved regions where the minority-phase domains localize and coalesce by decreasing the line energy through domain fusion. We show that this process is governed by the respective fraction of the two phases, and not by the chemical nature of the lipids involved. The spatial position and number of domains are controlled by the design of the confining microchannel and could result in polarized GUVs with a controllable number of poles. The developed method is versatile and user-friendly, while allowing multiple single-vesicle experiments in parallel.
巨型单层囊泡（GUVs）是广泛应用的最小细胞模型，在其中可以重现、分离和研究基本的生物学特征。尽管膜结构域精确的时空分布在活细胞中是一个至关重要的过程，但要生成具有位于用户定义位置的结构域的各向异性GUVs仍然极具挑战性。在此，我们描述了一种新颖且稳健的方法，用于控制巨型单层囊泡中液相有序（Lo）/液相无序（Ld）相的脂质结构域的空间位置。我们的策略是将发生Lo/Ld相分离的GUVs限制在微流控通道中，以定义自由弯曲区域，通过域融合降低线能量，少数相结构域在该区域定位并合并。我们表明，这一过程由两相各自的比例决定，而非由所涉及脂质的化学性质决定。结构域的空间位置和数量由限制微通道的设计控制，并且可以产生具有可控极数的极化GUVs。所开发的方法通用性强且用户友好，同时允许并行进行多个单囊泡实验。
