优化纳米乳剂在活细胞中单粒子追踪的荧光性质,ACS Applied Materials & Interfaces

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优化纳米乳剂在活细胞中单粒子追踪的荧光性质
ACS Applied Materials & Interfaces ( IF 8.3 ) Pub Date : 2019-03-07 00:00:00 , DOI: 10.1021/acsami.8b22297
Xinyue Wang ₁ , Nicolas Anton ₁ , Pichandi Ashokkumar ₂ , Halina Anton ₂ , Tkhe Kyong Fam ₂ , Thierry Vandamme ₁ , Andrey S. Klymchenko ₂ , Mayeul Collot ₂

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纳米乳液（NEs）是生物相容的脂质纳米颗粒，由表面活性剂壳稳定的油性核组成。公认的是，通过使用非离子表面活性剂，用聚乙二醇进行的表面装饰使生物介质具有较高的隐身性，并减少了非特异性相互作用。通过荧光显微镜技术跟踪单个NE将导致人们更好地了解它们在细胞中的行为，因此需要开发出具有增强光稳定性的明亮的单个颗粒。然而，一方面对NE的理化性质和化学组成之间的关系以及另一方面对被包封的染料的荧光性质之间的关系的理解仍然是有限的。在此处，^–1 cm ^–1）在溶剂中的量子产率相对较低，当溶解在粘性油中时，荧光增强效果显着（最高0.98）。据报道，对9种不同的油进行了筛选，从而可以揭示一系列有效的“油/染料”对，并了解导致NE最亮的主要参数。我们将维生素E醋酸盐/ DBS-C _8确定为最有效的代表，结合了高染料负载能力和低聚集诱导的猝灭作用，从而导致了<50 nm的超高亮NEs（亮度高达30×10 ⁶ M ^–1 cm ^–1）在生物介质中的染料泄漏可忽略不计。除了对荧光NE进行全面的光学和物理化学表征之外，还使用了聚合物涂层的细胞穿透性NE进行了细胞双光子激发成像。由于其令人印象深刻的亮度和光稳定性，将显示不同电荷表面的NE显微注射到HeLa细胞中，并在细胞溶胶中进行单独跟踪以研究其相对速度。

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Optimizing the Fluorescence Properties of Nanoemulsions for Single Particle Tracking in Live Cells

Nanoemulsions (NEs) are biocompatible lipid nanoparticles composed of an oily core stabilized by a surfactant shell. It is acknowledged that the surface decoration with poly(ethylene glycol), through the use of nonionic surfactants, confers high stealth in biological medium with reduced nonspecific interactions. Tracking individual NE by fluorescence microscopy techniques would lead to a better understanding of their behavior in cells and thus require the development of bright single particles with enhanced photostability. However, the understanding of the relationship between the physicochemical properties and chemical composition of the NEs, on the one hand, and its fluorescence properties of encapsulated dyes, on the other hand, remains limited. Herein, we synthesized three new dioxaborine barbituryl styryl (DBS) dyes that displayed high molar extinction coefficients (up to 120 000 M^–1 cm^–1) with relatively low quantum yields in solvents and impressive fluorescence enhancement when dissolved in viscous oils (up to 0.98). The reported screening of nine different oils allowed disclosing a range of efficient “oil/dye” couples and understanding the main parameters that lead to the brightest NEs. We determine vitamin E acetate/DBS-C₈ as the representative most efficient couple, combining high dye loading capabilities and low aggregation-induced quenching, leading to <50 nm ultrabright NEs (with brightness as high as 30 × 10⁶ M^–1 cm^–1) with negligible dye leakage in biological media. Beyond a comprehensive optical and physicochemical characterization of fluorescent NEs, cellular two-photon excitation imaging was performed with polymer-coated cell penetrating NEs. Thanks to their impressive brightness and photostability, NEs displaying different charge surfaces were microinjected in HeLa cells and were individually tracked in the cytosol to study their relative velocity.

更新日期：2019-03-07

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