使用單一平台即時細胞生長記錄儀追踪STO細胞株氧化壓力損傷後的凋亡狀態

Tracking the state of oxidative stress damage in STO cell line using a single platform of real-time cell growth recorder

龐鈞庭(Chun-Ting Pang)

指導教授 ： 張格東

國立屏東科技大學/農學院/生物科技系/碩士(2019年)

https://doi.org/10.6346/NPUST201900409

摘要

本研究藉由單一平台儀器系統的研發，以增進體外試驗細胞模型實驗方式之儀器為開發目的，藉由直接於培養環境下的即時影像拍攝，觀察並紀錄細胞型態、細胞表型之方式進行實驗觀察分析。
本次實驗模型以日常生活中，皮膚細胞所常見之氧化傷害；皮膚細胞易受太陽光中UV光照射造成氧化傷害，進而導致細胞損傷、生長停滯、細胞凋亡、DNA傷害，而藉由本研究研發之儀器以不同濃度H2O2氧化傷害之處理，並使用螢光染劑DCFH-DA、pSIVA觀察其螢光反應變化，再使用由互補性氧化金屬半導體（Complementary Metal-Oxide Semiconductor，CMOS）以及光學系統設計之儀器，以明視野（light field）、暗視野（Dark field）的搭配應用，並利用縮時攝影（Time-Lapse Photography）之概念，透過圖控程式NI LabVIEW進行控制定時拍攝，最後再由NI LabVIEW進行影像分析（二值化）。

關鍵字

氧化傷害 ； ROS ； CMOS ； 縮時攝影

並列摘要

In this study, the development of a single platform instrument system for improving the in vitro experimental model through observing and recording the image of cell morphology and phenotype directly in the culture environment.
In daily life, the oxidation of skin cells are susceptible to the UV light from sunlight, thus causing cell damage, growth arrest, apoptosis, and DNA damage. In this research, the STO cells were treated with different concentrations of H2O2 to induced oxidation damage and observed the fluorescence reaction with fluorescent dyes（DCFH-DA）and pSIVA by using complementary metal-oxide semiconductor（CMOS）with optical system. This optical system design instrument used a combination of light field and dark field, and using the concept of Time-lapse photography. Finally, the cell model were observed by timed shooting and quantified the image analysis（binarization）through the NI LabVIEW.

並列關鍵字

oxidative damage ； ROS ； CMOS ； time-lapse photography

參考文獻

Ben-Porath, I. and Weinberg, R.A., 2004. When cells get stressed: an integrative view of cellular senescence. The Journal of clinical investigation, 113(1), pp.8-20.

Bosch, R., Philips, N., Suárez-Pérez, J., Juarranz, A., Devmurari, A., Chalensouk-Khaosaat, J. and González, S., 2015. Mechanisms of photoaging and cutaneous photocarcinogenesis, and photoprotective strategies with phytochemicals. Antioxidants, 4(2), pp.248-268.

Cai, C., Guo, Z., Yang, Y., Geng, Z., Tang, L., Zhao, M., Qiu, Y., Chen, Y. and He, P., 2016. Inhibition of hydrogen peroxide induced injuring on human skin fibroblast by Ulva prolifera polysaccharide. International journal of biological macromolecules, 91, pp.241-247.

Chen, Y. and Junger, W.G., 2012. Measurement of oxidative burst in neutrophils. In Leucocytes (pp. 115-124). Humana Press.

Dodig, S., Čepelak, I. and Pavić, I., 2019. Hallmarks of senescence and aging. Biochemia Medica, 29(3).