New fluorescent probes for the measurement of cell membrane viscosity
New fluorescent probes for the measurement of cell membrane viscosity
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DOI：
10.1016/s1074-5521(00)90061-9
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发表时间：
2001-02-01
影响因子：
--
通讯作者：
Theodorakis, EA
中科院分区：
文献类型：
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作者：
Haidekker, MA;Ling, TT;Theodorakis, EA
背景资料：分子转子是表现出粘度依赖性荧光量子产率的荧光分子，潜在地允许直接测量培养细胞中的细胞膜粘度。然而，可商购获得的转子不仅染色细胞膜，而且还结合微管蛋白并迁移到细胞质中。我们合成的分子相关的9-（二氰基乙烯基）-久洛利定（DCVJ），其特点是不同长度的烃链，以增加membrane compatibility. Results：较长的烃链连接到荧光转子减少迁移到细胞质和细胞的内部隔间的染料。已知降低膜粘度的对流体剪切应力的荧光响应的幅度显著高于从DCVJ获得的响应。值得注意的是，法呢基链表现出超过20倍的幅度DCVJ和允许检测膜粘度的变化，在显着较低的剪切stresss.Conclusions：对增加细胞膜协会的分子转子的修改提供了一个新的研究工具，膜粘度测量。这些转子的使用补充了已建立的方法，例如光漂白后的荧光恢复，其具有有限的空间和时间分辨率以及荧光各向异性，其具有低灵敏度并且可能受到其他影响，例如变形。(C)2001爱思唯尔科技有限公司版权所有。
Background: Molecular rotors are fluorescent molecules that exhibit viscosity-dependent fluorescence quantum yield, potentially allowing direct measurements of cell membrane viscosity in cultured cells. Commercially available rotors, however, stain not only the cell membrane, but also bind to tubulin and migrate into the cytoplasm. We synthesized molecules related to 9-(dicyanovinyl)-julolidine (DCVJ), which featured hydrocarbon chains of different length to increase membrane compatibility.Results: Longer hydrocarbon chains attached to the fluorescent rotor reduce the migration of the dye into the cytoplasm and internal compartments of the cell. The amplitude of the fluorescence response to fluid shear stress, known to decrease membrane viscosity, is significantly higher than the response obtained from DCVJ. Notably a farnesyl chain showed a more than 20-fold amplitude over DCVJ and allowed detection of membrane viscosity changes at markedly lower shear stresses.Conclusions: The modification of molecular rotors towards increased cell membrane association provides a new research tool for membrane viscosity measurements. The use of these rotors complements established methods such as fluorescence recovery after photobleaching with its limited spatial and temporal resolution and fluorescence anisotropy, which has low sensitivity and may be subject to other effects such as deformation. (C) 2001 Elsevier Science Ltd. All rights reserved.