Abstract

Objective: Lipid droplets (adiposomes) are cellular organelles with distinguish structure and functions that play significant role in energy homeostasis and lipotoxicity and oxidative stress prevention, therefore represent a desirable target for fluorescent labeling. In this work, we proposed a number of new fluorogenic compounds—arylidene-rhodanines and their acyclic analogues – substances that label lipid droplets due to structural fragments responsible for the appearance of fluorescence in a lipophilic environment. Methods: The target arylidene-rhodanines and their acyclic analogues were obtained using modified literature methods of Knoevenagel condensation, the optical properties of the resulting compounds in a standard set of solvents were studied and then the most promising substances were tested as fluorescent dyes on Hela Kyoto cells using widefield microscopy. Discussion: We discovered that the derivatives containing a 2,5-dimethoxybenzylidene fragment or similar groups are characterized by a noticeable variation in fluorescence quantum yield depending on the environment properties. Cellular labeling of such compounds was studied. We showed that incubation of Hela Kyoto cells with substances (In) and (III) led to fluorescence appearance in the BFP channel, associated with the location of lipid droplets detected in the cells when visualized in transmitted light. Conclusions: As a result, we found that two obtained compounds—dimethoxybenzylidene-rhodanine and dimethoxybenzylidene-malononitrile – can be used as selective fluorogenic dyes for lipid droplets labeling in live Hela Kyoto cells under widefield fluorescent microscopy conditions.

中文翻译：

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2,5-二甲氧基-亚苄基-绕丹宁及其无环类似物作为活细胞脂滴的选择性荧光染料

摘要

目的：脂滴（脂肪体）是具有独特结构和功能的细胞器，在能量稳态、脂毒性和氧化应激预防中发挥重要作用，因此是荧光标记的理想目标。在这项工作中，我们提出了许多新的荧光化合物——亚芳基绕丹宁及其无环类似物——这些物质可以标记脂滴，因为结构片段负责在亲脂环境中出现荧光。方法：使用改进的Knoevenagel缩合文献方法获得目标亚芳基绕丹宁及其无环类似物，研究所得化合物在一组标准溶剂中的光学性质，然后在Hela京都上测试最有前途的物质作为荧光染料使用宽视野显微镜观察细胞。讨论：我们发现含有 2,5-二甲氧基苯亚甲基片段或类似基团的衍生物的特征是荧光量子产率根据环境特性而发生显着变化。研究了此类化合物的细胞标记。我们发现，用物质 ( In ) 和 ( III )孵育 Hela京都细胞会导致 BFP 通道中出现荧光，这与在透射光下观察时在细胞中检测到的脂滴位置相关。结论：结果，我们发现所获得的两种化合物——二甲氧基苯亚甲基-绕丹宁和二甲氧基苯亚甲基-丙二腈——可以用作选择性荧光染料，在宽场荧光显微镜条件下标记活 Hela京都细胞中的脂滴。