New Lipophilic Fluorescent Dyes for Labeling Extracellular Vesicles: Characterization and Monitoring of Cellular Uptake
New Lipophilic Fluorescent Dyes for Labeling Extracellular Vesicles: Characterization and Monitoring of Cellular Uptake
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DOI：
10.1021/acs.bioconjchem.1c00068
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发表时间：
2021-03-13
影响因子：
4.7
通讯作者：
Akiyoshi, Kazunari
中科院分区：
文献类型：
--
作者：
Shimomura, Takashi;Seino, Ryo;Akiyoshi, Kazunari
PKH染料是目前应用最广泛的细胞外囊泡（EV）标记荧光探针，但存在一定的局限性。例如，这些染料倾向于聚集，导致形成可以被细胞吸收的EV样纳米颗粒。此外，已经表明PKH染料由于膜融合或嵌入而引发EV的扩大。为了克服这些局限性，我们开发了三种新型的细胞外囊泡膜结合荧光探针。Mem染料-绿色、Mem染料-红色和Mem染料-深红-用于监测EV摄取进入细胞。该染料含有作为荧光支架的花青基团和花青上的两亲性部分。这三种染料具有不同的物理化学特性。为了研究用于EV标记的Mem染料的特性，我们进行了纳米颗粒跟踪、zeta电位测量和共聚焦显微镜。该染料能够实现EV的高灵敏度荧光成像。它们也可用于观察活细胞中的EV动力学。Mem染料显示出优异的EV标记，没有聚集和较少的颗粒扩大。
PKH dyes, which are currently the most widely used fluorescent probes for extracellular vesicle (EV) labeling, have some limitations. For example, these dyes tend to aggregate, leading to formation of EV-like nanoparticles that can be taken up by cells. Moreover, it has been suggested that PKH dyes trigger an enlargement of EVs because of membrane fusion or intercalation. To overcome these limitations, we developed three novel extracellular vesicular-membrane-binding fluorescent probes. Mem dye-Green, Mem dye-Red, and Mem dye-Deep Red-for monitoring EV uptake into cells. The dyes contain a cyanine group as a fluorescent scaffold and amphiphilic moieties on the cyanine. The three dyes have different photophysical characteristics. To investigate the characteristics of the Mem dyes for EV labeling, we performed nanoparticle tracking, zeta potential measurements, and confocal microscopy. The dyes enable highly sensitive fluorescence imaging of EVs. They can also be used to observe EV dynamics in live cells. The Mem dyes show excellent EV labeling with no aggregation and less particle enlargement.