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基于纳米粒子的生物相容性和长寿命标记物,用于溶酶体标记和追踪。

Nanoparticle-based biocompatible and long-life marker for lysosome labeling and tracking.

机构信息

State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Biomedical Engineering Center, Hunan University, Key Laboratory for Bio-Nanotechnology and Molecule Engineering of Hunan Province, Changsha 410082, China.

出版信息

Anal Chem. 2010 Mar 15;82(6):2213-20. doi: 10.1021/ac902417s.

Abstract

In this paper, a novel biocompatible and long-life lysosome labeling and tracking method based on dye entrapped silica nanoparticles (DSiNPs) has been put forward. Through colocalization studies using LysoTracker Green as the standard lysosome marker, it has been demonstrated that DSiNPs selectively accumulated in lysosomes of Hela cells and the photostability of DSiNPs associated with lysosomes was detectable, at least, 30 times as long as that of LysoTracker Green involved in lysosomes. By comparison with LysoTracker Green and Alexa 488-dextran, the fluorescence of DSiNPs could be detected over a 5-day postrecultivation period and the staining pattern in lysosomes could be well retained after cell fixation and permeabilization. In addition, results from MTT assays showed that DSiNPs did not affect the viability of Hela cells at the concentration for lysosome labeling. Primary applications of DSiNPs were then further performed in lysosome tracking in chloroquine-treated Hela cells, and lysosome labeling of differnet cell lines, including MCF-7 cells, MEAR cells, and MSC cells. These results indicated that DSiNPs, therefore, can be used as a biocompatible, long-life, and highly photostable lysosome marker for lysosome-related studies.

摘要

本文提出了一种新型的基于染料包埋二氧化硅纳米粒子(DSiNPs)的生物相容性和长寿命溶酶体标记和跟踪方法。通过使用 LysoTracker Green 作为标准溶酶体标记物的共定位研究,已经证明 DSiNPs 选择性地积累在 Hela 细胞的溶酶体中,并且与溶酶体相关的 DSiNPs 的光稳定性可检测到,至少比涉及溶酶体的 LysoTracker Green 长 30 倍。与 LysoTracker Green 和 Alexa 488-葡聚糖相比,DSiNPs 的荧光可以在培养后 5 天内检测到,并且在细胞固定和透化后,溶酶体中的染色模式可以很好地保留。此外,MTT 测定结果表明,在用于溶酶体标记的浓度下,DSiNPs 不会影响 Hela 细胞的活力。然后进一步将 DSiNPs 应用于氯喹处理的 Hela 细胞中的溶酶体跟踪以及不同细胞系(包括 MCF-7 细胞、MEAR 细胞和 MSC 细胞)的溶酶体标记。这些结果表明,DSiNPs 可以用作生物相容性、长寿命和高稳定性的溶酶体标记物,用于溶酶体相关研究。

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