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首页> 外文会议>Conference on Optical Diagnostics of Living Cells Ⅴ, Jan 23-25, 2002, San Jose, USA >Oxygen measurement in living cells: Comparison between a new vital fluorescent Pyrene probe labeling mitochondria and Pyrene Butyric Acid
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Oxygen measurement in living cells: Comparison between a new vital fluorescent Pyrene probe labeling mitochondria and Pyrene Butyric Acid

机译：活细胞中的氧气测量：标记线粒体的新型重要荧光vital探针与P丁酸之间的比较

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摘要

For the study of biological systems such as living cells, access to oxygen concentrations in various organelles is important. In living cells, the lifetime of pyrene fluorescence can be used to measure local oxygen concentrations. We designed and synthesized a new probe to measure the oxygen concentration in mitochondria ([1"-pyrene butyl]-2-rhodamine ester, PRE). The localization of the probe was studied by videomicrofluometry in 3T3 cells and confirmed to label the mitochondria. We compared the lifetime of PRE with the well-known cytosol specific probe Pyrene Butyric Acid in (ⅰ) in living cells (ⅱ) in liposomes and (ⅲ) in solution. Liposomes were used to investigate the effect of phospholipid bilayer organization on the fluorescence lifetimes. Depending on the oxygen concentration we observed lifetime variation ranges of (ⅰ) 60 ns (hyperoxygenation) ― 110 ns (anoxy) in cells (ⅱ) 60 ― 220 ns in liposomes and (ⅲ) 6 ― 220 ns in ethanolic solution. These results indicate that, under hyperoxygenation, quenching is less efficient in organized environments than in solution. Without oxygen and in cellular medium, the quenching depends on the composition of the probe environment. Accordingly, these probes can be used to measure the intracellular oxygen concentrations as well as changes in the environment.
机译：对于研究诸如活细胞的生物系统，获取各种细胞器中的氧气浓度很重要。在活细胞中，pyr荧光的寿命可用于测量局部氧浓度。我们设计并合成了一种用于测量线粒体中氧浓度的新探针（[1“ -py丁基] -2-若丹明酯，PRE），并通过视频显微技术在3T3细胞中研究了该探针的定位，并证实了其可标记线粒体。我们将PRE的寿命与著名的胞浆特异性探针probe丁酸在活细胞中的（ⅰ）在脂质体中的（and）和溶液中（in）的寿命进行了比较，并使用脂质体研究了磷脂双层结构对荧光的影响。根据氧浓度，我们观察到细胞的寿命变化范围为（ⅰ）60 ns（高氧）〜110 ns（无氧）（脂质体中）（60）60〜220 ns，在乙醇溶液中的（ⅲ）6〜220 ns。这些结果表明，在高氧条件下，淬灭在有组织的环境中比在溶液中效率低；在没有氧气和细胞培养基的情况下，淬灭取决于探针环境的组成，因此，这些探针可用于o测量细胞内的氧气浓度以及环境变化。

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