Cell Type Related Differences in Staining with Pentameric Thiophene Derivatives
Cell Type Related Differences in Staining with Pentameric Thiophene Derivatives
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DOI：
10.1002/cyto.a.22437
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发表时间：
2014-07-01
期刊：
影响因子：
3.7
通讯作者：
Los, Marek J.
中科院分区：
文献类型：
--
作者：
Cieslar-Pobuda, Artur;Back, Marcus;Los, Marek J.
能够选择性地染色细胞而不影响其生存能力的荧光化合物在生物学和医学中越来越重要。最近，一类新的光学染料，称为发光共轭低聚噻吩（LCOs），已成为一类有趣的高发射分子，用于研究各种生物现象。适当功能化的LCO已被用于选择性鉴定疾病相关蛋白质聚集体和选择性检测不同的细胞。在此，我们提供了各种细胞类型（包括癌细胞）的差异染色数据。用新开发的五聚体LCO观察到的差异染色归因于沿噻吩主链的不同侧链官能团沿着。采用流式细胞术和荧光显微镜，我们检查了一个图书馆的LCOs的多种细胞系的扩增。在测试的染料中，我们发现了有希望的候选物，其显示出强或中等的染色细胞到不同程度的能力，这取决于靶细胞。因此，沿噻吩主链沿着具有不同咪唑基序的LCO被鉴定为用于癌细胞染色的感兴趣的一类试剂，而沿主链沿着具有其它氨基酸侧链的LCO显示对研究中包括的细胞完全缺乏染色。此外，对于p-HTMI，一种用甲基化咪唑部分官能化的LCO，染色依赖于细胞的p53状态，表明染料的分子靶标是由p53调节的细胞组分。我们预见功能化的LCO将作为一类新的光学配体用于细胞的荧光分类，并扩展用于不同细胞的荧光活体成像的试剂工具箱。(C)2014年国际细胞计数促进学会
Fluorescent compounds capable of staining cells selectively without affecting their viability are gaining importance in biology and medicine. Recently, a new family of optical dyes, denoted luminescent conjugated oligothiophenes (LCOs), has emerged as an interesting class of highly emissive molecules for studying various biological phenomena. Properly functionalized LCOs have been utilized for selective identification of disease-associated protein aggregates and for selective detection of distinct cells. Herein, we present data on differential staining of various cell types, including cancer cells. The differential staining observed with newly developed pentameric LCOs is attributed to distinct side chain functionalities along the thiophene backbone. Employing flow cytometry and fluorescence microscopy we examined a library of LCOs for stainability of a variety of cell lines. Among tested dyes we found promising candidates that showed strong or moderate capability to stain cells to different extent, depending on target cells. Hence, LCOs with diverse imidazole motifs along the thiophene backbone were identified as an interesting class of agents for staining of cancer cells, whereas LCOs with other amino acid side chains along the backbone showed a complete lack of staining for the cells included in the study. Furthermore, for p-HTMI,a LCO functionalized with methylated imidazole moieties, the staining was dependent on the p53 status of the cells, indicating that the molecular target for the dye is a cellular component regulated by p53. We foresee that functionalized LCOs will serve as a new class of optical ligands for fluorescent classification of cells and expand the toolbox of reagents for fluorescent live imaging of different cells. (C) 2014 International Society for Advancement of Cytometry