Construction of Time-Lapse Scanning Electrochemical Microscopy with Temperature, Control and Its Application To Evaluate the Preservation Effects of Antifreeze Proteins on Living Cells
Construction of Time-Lapse Scanning Electrochemical Microscopy with Temperature, Control and Its Application To Evaluate the Preservation Effects of Antifreeze Proteins on Living Cells
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DOI：
10.1021/ac8018334
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发表时间：
2008-12-01
影响因子：
7.4
通讯作者：
Komatsu, Yasuo
中科院分区：
文献类型：
--
作者：
Hirano, Yu;Nishimiya, Yoshiyuki;Komatsu, Yasuo
抗冻蛋白（AFP）可以保护细胞免受低温损伤，但其作用机制尚不清楚。扫描电化学显微镜（SECM）可以评估细胞的大小和活性，尽管长期连续监测一直不成功。我们构建了一个新的，全自动的，时间推移SECM系统，并研究了AFP的细胞保存效果，通过分析单细胞地形在低温下。根据SECM测量，在4 ℃下在Euro-Collins（EC）溶液中处理的哺乳动物细胞（HepG 2）在8小时开始膨胀，然后立即破裂。在含AFP的EC溶液中，细胞大小直到16 h才发生变化，然后逐渐增大，最后破裂。此外，在AFF的存在下，破裂点处的细胞高度显著增加。这些结果表明，AFP稳定细胞膜，保护细胞免受低温损伤。这种SECM系统使我们能够通过长期自动扫描观察单个细胞对低温的反应，并将适用于分析其他细胞的活动和地形。
Antifreeze proteins (AFPs) can protect cells from hypothermic damage; however, their mechanism of action remains unclear. Scanning electrochemical microscopy (SECM) can evaluate the size and activities of cells, although long-term continuous monitoring has been unsuccessful. We constructed a novel, fully automated, timelapse SECM system and investigated the cell preservation effect of AFPs by analyzing single cellular topography at low temperatures. From the SECM measurements, mammalian cells (HepG2), treated in Euro-Collins (EC) solution at 4 degrees C, began to swell at 8 h and then immediately ruptured. In AFP-containing EC solution, the cellular size did not change until 16 h and then gradually increased and finally ruptured. In addition, the cellular height at rupture point significantly increased in the presence of AFF's. These results suggest that AFPs stabilize the cellular membrane and protect cells from hypothermic damage. This SECM system allowed us to observe the single cellular response to hypothermia by long-term automatic scanning and will be applicable for analysis to other cellular activities and topographies.