本實驗利用電化學交流阻抗法即時監測動物細胞貼附於電極上的行為，這項技術被稱為Electric Cell–substrate Impedance Sensing (ECIS)。檢測的項目為不同生長情況的小鼠纖維母細胞（L929）於電極上因懸浮或貼附所引起的阻抗值變化。本研究所使用的感測元件是利用雷射刻劃在耐高溫、耐化學藥品的聚醯亞胺(Polyimide)基板表面所製作出的石墨烯電極，稱為雷射誘導石墨烯(Laser-induced graphene, LIG)電極。電極型態上我們將選擇指叉電極；經過許多不同的電極尺寸、電極數量的設計與量測結果，最終將選擇考慮細胞大小以及LIG所能達到的尺寸之條件下期望得到穩定且靈敏度高的電極之尺寸。將小鼠纖維母細胞(L929)放置在指叉電極表面上的培養液中生長，使用低真空掃描式電子顯微鏡(LV-SEM)讓我們能夠清楚觀察在電極上面的增殖、生長、貼附的情形。監測上，利用電化學阻抗分析儀量L929細胞在含有不同濃度的毒化物或是藥品之培養液中生長時細胞和基底電極間的阻抗頻譜隨時間之變化。實驗結果顯示毒化物對細胞生長、貼附行為的影響可以利用測量LIG電極的阻抗頻譜而即時監控。本方法的優點包括可靠性高、分析速度快、精確性高，而且使用的樣品數和試劑少，就可以獲得相當好的樣品資訊。;This study used electrochemical AC (alternating-current) impedance method to monitor the behavior of animal cells attached to electrodes in real time, i.e., to conduct Electric Cell-substrate Impedance Sensing (ECIS). The target of detection was the impedance changes caused by suspension or attachment of mouse fibroblasts (L929) in different growth conditions on the interdigital electrodes. Laser-induced graphene (LIG) electrodes were used for measuring the cell-substrate impedance as well as for the substrate on which cells attached and grew. They were fabricated by laser scribing on the surface of heat-resistant, chemical-resistant polyimide film. Mouse fibroblasts (L929) were grown in culture medium on the surface of the interdigitated electrodes. Low-vaccume scanning electron microscopy (LV-SEM) allowed us to visually verify the cell proliferation, growth, and attachment on the electrodes. Then, the impedance spectrum between the cells and the substrate electrode was measured with time when L929 cells were grown in culture media containing different concentrations of poisons or drugs by electrochemical impedance analyzer. The experimental results showed that the feasibility of real-time monitoring the effects of poisons on cell growth and attachment behavior by measuring the cell-substrate impedance spectrum with LIG electrodes. The advantages of this method include high reliability, high analysis speed, high accuracy, and relatively good sample information with a small number of samples and reagents used.
