Imaging of Cell-Array by Scanning Electrochemical Microscopy

通过扫描电化学显微镜对细胞阵列进行成像

• 批准号: 13450348
• 负责人: MATSUE Tomokazu
• 金额: $ 9.47万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13450348/
• 关键词: Electrochemistry; Microscopy; Cell adhesion; Fibronectin; Patterning; Bioassay; Cardiac myocytes; Neuron; パターンニング

In the present project, we investigated two topics: 1) the micropatterning of living cells, 2) activity measurements of single cells. These research topics can be combined as the "bioassay using micropatterned living cells", which would lead to the development of cell-chip technology. The results obtained in this project can be summarized as follows,1) Preparation of cellular micropatterns: We succeeded to prepare micropatterns of adhesive animal cells on a glass slides by the micro-contact printing method using PDMS-made microstumps. The cellular adhesive protein such as fibronectin was used as the ink material. HeLa cells, chick cardiomyocytes and PC12 nuronal cells were successfully patterned to form cellular networks on a single cell level. The application of PEG molecules as the cell resistive modification works well so as to maintain the cell pattern for more than 5 days.2) Activity evaluation of the micropatterned cells: The respiratory activity of the micropatterned HeLa cells was imaged by using the scanning electrochemical microscopy (SECM) system. It was found that the respiration activity of the shape-restricted cells is higher than that of the freely spreading cells.3) Evaluation of the activity of the patterned cellular networks: The intracellular Ca2+ imaging was conducted for the micropatterned cardiac myocytes. The micropatterned cardiac myocytes were found to form the electrically conjugated network via forming the gap junctions. It was achieved to observed the pharmacological effects of octanol, tetrodotoxyn, and caffein for the cardiac tissue model on the chip. The local dosing to the patterned cells has also been attempted by applying the microfuluidic device systems.

在本项目中，我们研究了两个主题：1）活细胞的微图案，2）单细胞的活性测量。这些研究主题可以与“使用微图案活细胞的生物测定”合并，这将导致细胞芯片技术的发展。该项目中获得的结果可以总结如下：1）细胞微图案的制备：我们成功地通过使用PDMS制造的微块状的微接触方法在载玻片上制备了粘合动物细胞的微图案。细胞粘附蛋白（例如纤连蛋白）用作墨水材料。 HeLa细胞，雏鸡心肌细胞和PC12 NURONAL细胞已成功形成图案，以在单个细胞水平上形成细胞网络。将PEG分子作为细胞电阻修饰的应用很好地效果很好，以使细胞模式超过5天。2）微图案细胞的活性评估：通过使用扫描的电化学显微镜（SECM）系统对微图案HELA细胞的呼吸活性进行成像。已经发现，形状限制细胞的呼吸活性高于自由扩散的细胞的呼吸活性。3）对图案化细胞网络的活性进行评估：对微图案心肌细胞进行细胞内Ca2+成像。发现微图案心肌细胞通过形成间隙连接形成电偶联网络。观察到芯片上心脏组织模型的辛醇，四毒素和caffein的药理作用已达到实现。还尝试通过应用微纤维纤维设备系统来尝试对图案单元格的局部剂量。

项目成果

M.Nishizawa: "Micropatterning of Hela Cells on Glass Substrates and Evaluation of Respiratory Activity Using Microelectrodes"Langmuir. 18. 3645-3649 (2002)

M.Nishizawa：“玻璃基板上 Hela 细胞的微图案化和使用微电极评估呼吸活动”Langmuir。

H.Kaji: "Intracellular Ca^<2+> Imaging for Micropatterned Cardiac Myocytes"Biotech. Bioeng.. 81. 748-751 (2003)

H.Kaji：“微图案心肌细胞的细胞内 Ca^<2 > 成像”生物技术。

Y.Hirano: "Microspots of GOD-HRP Bienzyme for Scanning Chemiluminescence Microscopy with Higher Resolution"Electrochemistry. 68. 946-948 (2001)

Y.Hirano：“用于高分辨率扫描化学发光显微镜的 GOD-HRP 双酶微点”电化学。

M.Nishizawa: "Micropatterning of HeLa Cells on Glass Substrates and Evaluation of Respiratory Activity Using Microelectrodes"Langmuir. 印刷中. (2002)

M.Nishizawa：“玻璃基板上 HeLa 细胞的微图案化和使用微电极评估呼吸活动”Langmuir，出版。

Matsuhiko Nishizawa, Kimiyasu Takoh, Tomokazu Matsue: "Micropatterning of HeLa Cells on Glass Substrates and Evaluation of Respiratory Activity Using Microelectrodes"Langmuir. 18. 3645-3649 (2002)

Matsuhiko Nishizawa、Kimiyasu Takoh、Tomokazu Matsue：“玻璃基板上 HeLa 细胞的微图案化和使用微电极评估呼吸活动”Langmuir。