Tucker K G, Chalder S, al-Rubeai M, Thomas C R
SERC Centre for Biochemical Engineering, School of Chemical Engineering, University of Birmingham, UK.
Enzyme Microb Technol. 1994 Jan;16(1):29-35. doi: 10.1016/0141-0229(94)90106-6.
A novel automated image analysis method is described for characterizing the viability and morphology of animal cells from suspension cultures. With the aid of the exclusion dye Trypan Blue, the total and viable cell counts and the percentage of dead cells present are found. The area, perimeter, equivalent diameter, and circularity of the projected image of each cell are also measured, allowing the estimation of cell volume. The image analysis method is inherently sensitive, precise, consistent (non-operator dependent) and relatively fast, taking approximately 22 min to analyze one sample. The data it gives on individual cellular activity, as characterized by Trypan Blue uptake and cell morphology, are valuable in allowing early diagnosis of even subtle changes in the health of a culture. The method should permit better optimization of culture conditions, and will provide data for the modeling of cell population dynamics. As the conventional manual method is operator dependent, relatively limited in the amount of information it provides, and has a tendency to underestimate Trypan Blue takeup, it is suggested that image analysis be the preferred option for animal cell counting and viability determinations.
本文描述了一种新型的自动图像分析方法，用于表征悬浮培养动物细胞的活力和形态。借助排斥染料台盼蓝，可得出总细胞数、活细胞数以及死细胞所占百分比。还测量了每个细胞投影图像的面积、周长、等效直径和圆形度，从而估算细胞体积。该图像分析方法本质上具有灵敏、精确、一致（不依赖操作人员）且相对快速的特点，分析一个样本大约需要22分钟。它给出的关于单个细胞活性的数据，以台盼蓝摄取和细胞形态为特征，对于早期诊断培养物健康状况中即使是细微的变化也很有价值。该方法应能更好地优化培养条件，并将为细胞群体动态建模提供数据。由于传统的手动方法依赖操作人员，提供的信息量相对有限，且有低估台盼蓝摄取量的倾向，因此建议图像分析是动物细胞计数和活力测定的首选方法。
