Boothby C D, Daniel J, Adam S, Dyson J E
University Department of Radiotherapy, Cookridge Hospital, Leeds, United Kingdom.
In Vitro Cell Dev Biol. 1989 Oct;25(10):946-50. doi: 10.1007/BF02624008.
Increasing use is being made of tumor cell lines cultured as cell aggregates (generally referred to as multicellular spheroids) in in vitro radiosensitivity and/or chemosensitivity tests. Conventional procedures for the determination of mean spheroid diameters for the construction of growth delay curves employ a microscope-image analyzer. However, this approach can prove excessively time consuming when large numbers of samples have to be measured. We have, therefore, been exploring the use of a Laser Diffraction Particle Sizer, the Malvern 2600 long bench model, for the measurement of mean spheroid diameter and size distribution. We report here a direct comparison between measurements carried out by the instrument and under the microscope. Also a comparison of growth curves for six cell lines constructed from measurements by the microscope and by the instrument. A number of factors that might affect the accuracy of spheroid diameter measurement by the instrument have been investigated: The effect of stirring to maintain the spheroids in suspension during measurement. Sampling error due to removal of a series of spheroid samples from culture flasks for measurement. Optimum numbers of scans to be carried out by the instrument to reach a constant value for mean diameter, and minimum SE of the mean.
在体外放射敏感性和/或化学敏感性测试中，越来越多地使用培养为细胞聚集体（通常称为多细胞球体）的肿瘤细胞系。用于构建生长延迟曲线的平均球体直径测定的传统方法采用显微镜图像分析仪。然而，当必须测量大量样品时，这种方法可能会非常耗时。因此，我们一直在探索使用激光衍射粒度分析仪（马尔文2600长台型）来测量平均球体直径和尺寸分布。我们在此报告该仪器与显微镜下测量结果的直接比较。还比较了通过显微镜和该仪器测量构建的六种细胞系的生长曲线。已经研究了一些可能影响该仪器测量球体直径准确性的因素：测量期间搅拌以保持球体悬浮的影响。从培养瓶中取出一系列球体样品进行测量时的采样误差。仪器进行扫描的最佳次数，以达到平均直径的恒定值和平均值的最小标准误差。
