細胞株可以提供細胞產物，基因學，細胞間交互作用，環境影響，胞內流量以及胞內活性的研究，而PC12細胞株已被廣泛用來研究了解神經凋亡，神經化學以及神經分化等領域。因此建構一合適兼具細胞培養控制平台與影像擷取分析功能之系統是相當重要研究課題。本研究的目的就是要建立一低成本的細胞培養系統，此系統包含顯微鏡影像分析系統，可以在細胞培養環境下同時擷取細胞影像加以處理分析。實驗結果發現，細胞培育系統在溫度控制方面可以達到37.3 ± 1.3℃，在相對濕度方面可以達到89.7 ± 1.8%。二氧化碳濃度的值為5.29 ± 0.71%。經過48小時後的培養，細胞數量由1.00 x 105 cells/ml上升到4.25 x 105 cells/ml。從細胞存活率測試證明本細胞培養系統在PC12細胞培養與分析有很好應用性；同時也可以在細胞培養過程中，隨時擷取影像並加以分割；本研究應用區域成長與區域標識方法來分割在培養條件下之PC12細胞，以利後續形態與特徵量化分析。

Cell line can be applied to provide the cell products, genetics, cell-cell interaction, environmental interaction, intracellular flux, and intracellular activity studies. The PC12 cell line is widely used in vitro study for investigating the neuronal apoptosis, neurochemistry, and neuronal differentiation. Therefore, the development of an appropriate cell cultivation platform as well as the real-time image acquisition and analysis system becomes an important issue. The purpose of this study is to develop a low-cost incubation system including microscopic image analysis system. The proposed system can acquire cell image for analysis during cell cultivation. From the experimental results, the temperature of incubation system ranges from 37.3 ± 1.3℃, and the lower bound of the relative humidity ranges from 89.7 ± 1.8%. The CO2 concentration is controlled at 5.29 ± 0.71%. The cell number is increased from 1.00 x 105 to 4.25 x 105 cells/ml after 48 hours cultivation. The cell viability test shows that the self-developed incubation system works well for PC12 cells cultivation; it can also acquire and segment the cell image during the cell cultivation. Besides, the region growing and modified blob labeling techniques are also employed to segment the PC12 cell image under cultivation conditions for later the morphology and feature quantification analysis.

[ 1 ] Http://www.neurologychannel.com/stroke/
[ 2 ] Http://www.nucleusinc.com
[ 3 ] J. A. Zivin, and U. DeGirolami, “Spinal cord infraction: a highly reproducible stroke model,” Stroke, vol. 11, no. 2, pp. 200-202, 1980.
[ 4 ] S. T. Chen, C. Y. Hsu, E. L. Hogan, H. Maricq, and J. D. Balentine, “A model of focal ischemic stroke in the rat: reproducible extensive cortical infarction,” Stroke, vol. 17, no. 4, pp. 738-743, 1986.
[ 5 ] A. Alemida, M. Delgado-Esteban, J. P. Bola s, and J. M. Medina, “Oxygen and glucose deprivation induces mitochondrial dysfunction and oxidative stress in neurons but not in astrocytes in primary culture,” Journal of Neurochemistry, vol. 81, pp. 207-217, 2002.
[ 6 ] A.C. Yu, G.A. Gregory, and P.H. Chan, “Hypoxia-induced dysfunctions and injury of astrocytes in primary cell cultures,” Journal of Cerebral Blood Flow and Metabolism, vol. 9, no. 1, pp. 20-28, February, 1989.
[ 7 ] P. H. Lee, E. M. Hwang, and H. S. Hong, “Effect of ischemic insults on amyloid precursor protein processing,” Neurochemical Research, vol. 31, pp. 821-827, June, 2006.
[ 8 ] R. I. Freshney, Culture of Animal Cells. New York: Wiley, 2000.
[ 9 ] S. L. Kroll, and M. F. Czyzyk-Krzeska, “Role of H2O2 and heme-containing O2 sensor in hypoxia regulation of tyrosine hydroxylase gene expression,” American Journal of Physiology, vol. 274, pp. 167-174,1998.

[10] L. A. Greene, and A. S. Tischler, “Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor,” Proceedings of the National Academy of Sciences, vol. 73, no. 7, pp. 2424-2428, July, 1976.
[11] E. Gozal, L. R. Sachleben, M. J. Rane, C. Vega, and D. Gozal, “Mild sustained and intermittent hypoxia induce apoptosis in PC-12 cells via different mechanisms,” American Journal of Physiology - Cell Physiology, vol. 288, pp. 535–542, 2005.
[12] Http://www.atcc.org/Home.cfm
[13] J. Saura, G. MacGibbon, and M. Dragunow, “Etoposide-induced PC12 cell death: apoptotic morphology without oligonucleosomal DNA fragmentation or dependency upon de novo protein synthesis,” Molecular Brain Research, vol. 48, pp. 382-388, 1997.
[14] T. Noma, Y. S. Yoon, and A. Nakazawa, “Overexpression of NeuroD in PC12 cells alters morphology and enhances expression of the adenylate kinase isozyme 1 gene,” Molecular Brain Research, vol. 67, pp. 53-63, 1999.
[15] T. T. Rohn, S. M. Cusack, S. R. Kessinger, and J. T. Oxford, “Caspase activation independent of cell death is required for proper cell dispersal and correct morphology in PC12 cells,” Experimental Cell Research, vol. 295, pp. 215-225, 2004.
[16] Z. Mi, Z. K. Mirnics, and N. F. Schor, “Bcl-2 overexpression disrupts the morphology of PC12 cells through reduced ERK activation,” Brain research, vol. 1112, pp. 46-55, 2006.
[17] E. Gozal, L. R. Sachleben, M. J. Rane, C. Vega, and D. Gozal, “Mild sustained and intermittent hypoxia induce apoptosis in PC-12 cells via different mechanisms,” American Journal of Physiology - Cell Physiology, vol. 288, pp. 535–542, 2005.
[18] D. J. Stephens, and V. J. Allan, “Light microscopy techniques for live imaging,” American Association for the Advancement of Science, vol. 300, pp. 82-86, 2003.
[19] C. D. Ruberto, A. Dempster, S. Khan, and B. Jarra, “Analysis of infected blood cell images using morphological operators,” Image and Vision Computing, vol. 20, pp. 133-146, 2002
[20] R. C. Gonzalez and R. E. Woods, Digital Image Processing. New Jersey: Prentice-Hall, 2002.
[21] A. E. Huque, Shape analysis and measurement for the HeLa cell classification of cultured cells in high throughput screening. Master Thesis, School of Humanities & Informatics University of Skövde, Sweden, February, 2006.
[22] L. P. Lin, Microscopy for Microorganism. Taiwan, Taipei: Yi-Shien, 2002 (in Chinese).
[23] D. P. H der, Image Analysis Method and Application. USA: CRC Press LLC, 2001