Glycosphingolipids, including gangliosides, are representative lipid raft markers that perform a variety of physiological roles in cell membranes. However, studies aimed at revealing their dynamic behavior in living cells are rare, mostly due to a lack of suitable fluorescent probes. Recently, the ganglio-series, lacto-series, and globo-series glycosphingolipid probes, which mimic the behavior of the parental molecules in terms of partitioning to the raft fraction, were developed by conjugating hydrophilic dyes to the terminal glycans of glycosphingolipids using state-of-art entirely chemical-based synthetic techniques. High-speed, single-molecule observation of these fluorescent probes revealed that gangliosides were scarcely trapped in small domains (100 nm in diameter) for more than 5 ms in steady-state cells, suggesting that rafts including gangliosides were always moving and very small. Furthermore, dual-color, single-molecule observations clearly showed that homodimers and clusters of GPI-anchored proteins were stabilized by transiently recruiting sphingolipids, including gangliosides, to form homodimer rafts and the cluster rafts, respectively. In this review, we briefly summarize recent studies, the development of a variety of glycosphingolipid probes as well as the identification of the raft structures including gangliosides in living cells by single-molecule imaging.

鞘糖脂，包括神经节苷脂，是代表性的脂筏标记物，在细胞膜中发挥多种生理作用。然而，旨在揭示它们在活细胞中的动态行为的研究很少见，这主要是由于缺乏合适的荧光探针。最近，神经节系列、乳糖系列和 globo 系列糖鞘脂探针通过使用状态-将亲水性染料与糖鞘脂的末端聚糖结合来开发，它们模拟亲本分子在分配到筏部分方面的行为。最先进的完全基于化学的合成技术。对这些荧光探针的高速单分子观察表明，在稳态细胞中，神经节苷脂几乎不会被困在小区域（直径 100 nm）中超过 5 毫秒，这表明包含神经节苷脂的筏总是在移动并且非常小。此外，双色、单分子观察清楚地表明，同二聚体和 GPI 锚定蛋白簇通过瞬时募集鞘脂（包括神经节苷脂）分别形成同二聚体筏和簇筏来稳定。在这篇综述中，我们简要总结了最近的研究、各种糖鞘脂探针的开发以及通过单分子成像识别活细胞中的筏结构，包括神经节苷脂。分别形成同源二聚体筏和簇筏。在这篇综述中，我们简要总结了最近的研究、各种糖鞘脂探针的开发以及通过单分子成像识别活细胞中的筏结构，包括神经节苷脂。分别形成同源二聚体筏和簇筏。在这篇综述中，我们简要总结了最近的研究、各种糖鞘脂探针的开发以及通过单分子成像识别活细胞中的筏结构，包括神经节苷脂。