Dojindo Laboratories , Tabaru 2025-5 , Mashiki-machi , Kumamoto 861-2202 , Japan.
Laboratory of Biochemistry and Molecular Biology, Department of Medical Sciences , University of Miyazaki , 5200 Kihara Kiyotake , Miyazaki 889-1692 , Japan.
Biochemistry. 2019 Feb 12;58(6):499-503. doi: 10.1021/acs.biochem.8b01071. Epub 2019 Jan 11.
We have developed three types of lipid droplet (LD)-specific fluorescent probes for live-cell imaging, Lipi-Blue, Lipi-Green, and Lipi-Red, which exhibit fluorescence upon being incorporated into LDs both of living and of fixed cells. These Lipi-probes are LD-specific probes that contain a pyrene or perylene group as a fluorescent scaffold and can be used to observe dynamics of LD in live cells and also interrelations with other organelles by simultaneous staining with multiple organelle-specific probes. Additionally, Lipi-Blue and Lipi-Green allow monitoring LDs in live cells even for 48 h after the staining. Here we show that newly formed LDs and previously existed LDs can be separately monitored in a single cell by using these probes and that intercellular transfer of whole LDs is observed in KB cells, but not in HepG2 cells under the same culturing condition. These findings indicate that newly developed LD-specific probes are useful to analyze the dynamics of LDs in live cells.
我们开发了三种用于活细胞成像的脂滴（LD）特异性荧光探针，分别为 Lipi-Blue、Lipi-Green 和 Lipi-Red，它们在被活细胞和固定细胞的 LD 内化后会发出荧光。这些 Lipi 探针是 LD 特异性探针，含有芘或苝作为荧光支架，可用于观察活细胞中 LD 的动态，以及通过与多个细胞器特异性探针同时染色来观察与其他细胞器的相互关系。此外，Lipi-Blue 和 Lipi-Green 允许在染色后 48 小时内仍在活细胞中监测 LD。在这里，我们表明，通过使用这些探针可以在单个细胞中分别监测新形成的 LD 和先前存在的 LD，并且可以观察到 KB 细胞中整个 LD 的细胞间转移，但在相同培养条件下 HepG2 细胞中则没有观察到。这些发现表明，新开发的 LD 特异性探针可用于分析活细胞中 LD 的动态。
