Suwara Justyna, Lukasik Beata, Zurawinski Remigiusz, Pawlowska Roza, Chworos Arkadiusz
Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Sienkiewicza 112, 90-363 Lodz, Poland.
Materials (Basel). 2020 Feb 20;13(4):951. doi: 10.3390/ma13040951.
Fluorescent imaging, which is an important interdisciplinary field bridging research from organic chemistry, biochemistry and cell biology has been applied for multi-dimensional detection, visualization and characterization of biological structures and processes. Especially valuable is the possibility to monitor cellular processes in real time using fluorescent probes. In this work, conjugated oligoelectrolytes and neutral derivatives with the distyrylnaphthalene core (SN-COEs) were designed, synthetized and tested for biological properties as membrane-specific fluorescent dyes for the visualization of membrane-dependent cellular processes. The group of tested compounds includes newly synthesized distyrylnaphthalene derivatives (DSNNs): a trimethylammonium derivative (DSNN-NMe), a phosphonate derivative (DSNN-P), a morpholine derivative (DSNN-Mor), a dihydroxyethylamine derivative (DSNN-DEA), a phosphonate potassium salt (DSNN-POK), an amino derivative (DSNN-NH) and pyridinium derivative (DSNN-Py+). All compounds were tested for their biological properties, including cytotoxicity and staining efficiency towards mammalian cells. The fluorescence intensity of SN-COEs incorporated into cellular structures was analyzed by fluorescence activated cell sorting (FACS) and photoluminescence spectroscopy. The cytotoxicity results have shown that all tested SN-COEs can be safely used in the human and animal cell studies. Fluorescence and confocal microscopy observations confirm that tested COEs can be applied as fluorescent probes for the visualization of intracellular membrane components in a wide range of different cell types, including adherent and suspension cells. The staining procedure may be performed under both serum free and complete medium conditions. The presented studies have revealed the interesting biological properties of SN-COEs and confirmed their applicability as dyes for staining the membranous structures of eukaryotic cells, which may be useful for visualization of wide range of biological processes dependent of the extra-/intracellular communications and/or based on the remodeling of cellular membranes.
荧光成像作为一个连接有机化学、生物化学和细胞生物学研究的重要跨学科领域，已被应用于生物结构和过程的多维检测、可视化和表征。使用荧光探针实时监测细胞过程的可能性尤其有价值。在这项工作中，设计、合成了具有二苯乙烯基萘核心的共轭寡电解质和中性衍生物（SN-COEs），并测试了它们作为膜特异性荧光染料的生物学特性，用于可视化依赖于膜的细胞过程。测试的化合物组包括新合成的二苯乙烯基萘衍生物（DSNNs）：三甲基铵衍生物（DSNN-NMe）、膦酸酯衍生物（DSNN-P）、吗啉衍生物（DSNN-Mor）、二乙醇胺衍生物（DSNN-DEA）、膦酸钾盐（DSNN-POK）、氨基衍生物（DSNN-NH）和吡啶鎓衍生物（DSNN-Py+）。所有化合物都测试了其生物学特性，包括对哺乳动物细胞的细胞毒性和染色效率。通过荧光激活细胞分选（FACS）和光致发光光谱分析了掺入细胞结构中的SN-COEs的荧光强度。细胞毒性结果表明，所有测试的SN-COEs都可安全用于人和动物细胞研究。荧光和共聚焦显微镜观察证实，测试的COEs可作为荧光探针，用于可视化包括贴壁细胞和悬浮细胞在内的多种不同细胞类型中的细胞内膜成分。染色过程可以在无血清和完全培养基条件下进行。所呈现的研究揭示了SN-COEs有趣的生物学特性，并证实了它们作为染料用于染色真核细胞膜结构的适用性，这可能有助于可视化广泛的依赖于细胞外/细胞内通讯和/或基于细胞膜重塑的生物学过程。
