Dual-Activatable Cell Tracker for Controlled and Prolonged Single-Cell Labeling.,ACS Chemical Biology

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Dual-Activatable Cell Tracker for Controlled and Prolonged Single-Cell Labeling.
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2020-04-16 , DOI: 10.1021/acschembio.0c00208
Elias A Halabi ₁ , Jorge Arasa ₂ , Salome Püntener _{1,

3} , Victor Collado-Diaz ₂ , Cornelia Halin ₂ , Pablo Rivera-Fuentes _{1,

3}

Affiliation

Cell trackers are fluorescent chemical tools that facilitate imaging and tracking cells within live organisms. Despite their versatility, these dyes lack specificity, tend to leak outside of the cell, and stain neighboring cells. Here, we report a dual-activatable cell tracker for increased spatial and temporal staining control, especially for single-cell tracking. This probe overcomes the typical problems of current cell trackers: off-target staining, high background signal, and leakage from the intracellular medium. Staining with this dye is not cytotoxic, and it can be used in sensitive primary cells. Moreover, this dye is resistant to harsh fixation and permeabilization conditions and allows for multiwavelength studies with confocal microscopy and fluorescence-activated cell sorting. Using this cell tracker, we performed in vivo homing experiments in mice with primary splenocytes and tracked a single cell in a heterogeneous, multicellular culture environment for over 20 h. These experiments, in addition to comparative proliferation studies with other cell trackers, demonstrated that the signal from this dye is retained in cells for over 72 h after photoactivation. We envision that this type of probes will facilitate the analysis of single-cell behavior and migration in cell culture and in vivo experiments.

中文翻译：

可双重激活的细胞追踪器，用于受控和长时间的单细胞标记。

细胞追踪器是荧光化学工具，可促进对活生物体内的细胞进行成像和追踪。尽管它们具有通用性，但这些染料缺乏特异性，易于泄漏到细胞外部，并对邻近细胞染色。在这里，我们报告一个双重激活的细胞追踪器，用于增加空间和时间染色控制，尤其是对于单细胞追踪。该探针克服了当前细胞追踪器的典型问题：脱靶染色，高背景信号和细胞内培养基渗漏。用这种染料染色不会产生细胞毒性，可用于敏感的原代细胞。此外，这种染料可抵抗苛刻的固定和透化条件，并允许通过共聚焦显微镜和荧光激活细胞分选技术进行多波长研究。使用此单元格跟踪器，我们执行了小鼠原代脾细胞的体内归巢实验，并在异质，多细胞培养环境中追踪单个细胞超过20小时。这些实验以及与其他细胞追踪剂的对比增殖研究表明，该染料的信号在光激活后在细胞中保留了72小时以上。我们设想这种类型的探针将有助于分析单细胞行为以及细胞培养和体内实验中的迁移。

更新日期：2020-06-19

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