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PKH67的无害性及细胞内分布:一种用于细胞增殖评估的荧光探针

Innocuousness and intracellular distribution of PKH67: a fluorescent probe for cell proliferation assessment.

作者信息

Rousselle C, Barbier M, Comte V V, Alcouffe C, Clement-Lacroix J, Chancel G, Ronot X

机构信息

Laboratoire de Dynamique Cellulaire, Ecole Pratique des Hautes Etudes, Université Joseph Fourier, La Tronche, France.

出版信息

In Vitro Cell Dev Biol Anim. 2001 Nov-Dec;37(10):646-55. doi: 10.1290/1071-2690(2001)037<0646:iaidop>2.0.co;2.

Abstract

PKH dyes were initially developed by Horan et al. to provide appropriate probes for in vitro and in vivo cell tracking. It has been reported for many cell types that PKH bind irreversibly to the cell membrane without significantly affecting cell growth. Thus, these probes provide an opportunity for long-term cell monitoring and the identification of cells of interest among a heterogeneous cell population. An important feature is that upon cell division, the probe is partitioned equally between each daughter cell, making it possible to quantify tell fluorescence by flow cytometry. In this situation. the flow cytometric study of PKH67 characteristics shows that this probe does not affect the main cell-functions such as viability or proliferation. Moreover, the intracellular distribution of PKH67 is demonstrated by following its kinetics of internalization by confocal microscopy. These results present PKH67 as a probe suitable for dynamic analysis of cell proliferation as well as the study of intracellular localization and membrane recycling mechanisms.

摘要

PKH染料最初由霍兰等人开发,用于提供适用于体外和体内细胞追踪的探针。据报道,对于许多细胞类型,PKH不可逆地结合到细胞膜上,而不会显著影响细胞生长。因此,这些探针为长期细胞监测以及在异质细胞群体中识别感兴趣的细胞提供了机会。一个重要特征是,在细胞分裂时,探针会在每个子细胞之间平均分配,从而可以通过流式细胞术对荧光进行定量分析。在这种情况下,对PKH67特性的流式细胞术研究表明,该探针不会影响诸如活力或增殖等主要细胞功能。此外,通过共聚焦显微镜追踪PKH67的内化动力学,证明了其在细胞内的分布。这些结果表明PKH67是一种适用于细胞增殖动态分析以及细胞内定位和膜循环机制研究的探针。

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