Chang C H, Lin C P, Wang H Z
Department of Ophthalmology, Kaohsiung Medical College, Taiwan, Republic of China.
Gaoxiong Yi Xue Ke Xue Za Zhi. 1993 Nov;9(11):610-5.
A tetrazolium-based(MTT) assay was used to evaluate the viability of cultured porcine monolayer corneal endothelial cells. In this study, we tried to find out the best spectra at which scanning multiwell ELISA spectrophometer can obtain the widest range of absorbance (optical density, O.D.) and the ranges of numbers of corneal endothelial cells in which absorbance is in a significant linear relationship with cell numbers. Among the three tested spectra of 450, 540, and 630nm, the widest range of absorbance was at 540 nm, from 0.1193 to 0.6183. At all three tested spectra, numbers of corneal endothelial cells from 50 to 30,000 were in a significant linear relationship with absorbance. Therefore, in the future studies to assay the viability of corneal endothelial cells, we shall choose the spectrum of 540 nm for reading with a scanning multiwell ELISA spectrophotometer, and absorbance(O.D.) from 0.1193 to 0.6183 will be accepted, representing cell numbers from 50 to 30,000.
采用基于四氮唑的（MTT）分析法评估培养的猪单层角膜内皮细胞的活力。在本研究中，我们试图找出扫描多孔酶标仪能获得最宽吸光度范围（光密度，O.D.）的最佳光谱，以及吸光度与细胞数量呈显著线性关系的角膜内皮细胞数量范围。在450、540和630nm这三个测试光谱中，540nm处的吸光度范围最宽，为0.1193至0.6183。在所有三个测试光谱下，50至30000个角膜内皮细胞的数量与吸光度呈显著线性关系。因此，在未来检测角膜内皮细胞活力的研究中，我们应选择540nm光谱用扫描多孔酶标仪进行读数，吸光度（O.D.）在0.1193至0.6183之间将被接受，代表50至30000个细胞数量。
