Imaging of Cell-Array by Scanning Electrochemical Microscopy

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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.