| Summary: | 碩士 === 國立成功大學 === 醫學工程研究所碩博士班 === 96 === The PC12 cell line has been widely used in stroke related in vitro study, due to its features of neuronal-like morphology and cell-like phenomenon under O2 deprived condition. The purposes of this study are to develop a real-time culturing conditions monitoring system for PC12 application, and to investigate the useful parameters for characterizing the types of cell death with no immunoassay labeling. From the experimental results, it is shown that the temperature, the mean relative humidity, the O2 concentration (5%), and the CO2 concentration (5%) for the proposed culturing system are controlled to be 37.1 ± 0.5℃, 95%, 4.88 ± 0.3%, and 5.72 ± 0.5%, respectively. For the cell image analysis, the feature parameters of the N/C ratio, area, and entropy are realized and applied to quantify the membrane and nucleus of PC12. From the experimental results of single cell continuously monitoring, the apoptosis or necrosis of a cell is differentiated using the thresholds of parameters of the entropy (0.29) and mean gray value (0.26) of membrane, area of nucleus (0.16), and N/C ratio (0.14) after being cultured for 13 to 19 hours. From the experimental results of multiple cells observations, it is shown that the OGD (5% O2) and lower hypoxic (1% O2) conditions would damage the cells more than that of mild hypoxic (5% O2) conditions.
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