Lundholt Betina Kerstin, Scudder Kurt M, Pagliaro Len
BioImage A/S, Moerkhoej Bygade 28, DK-2860 Soeborg, Denmark.
J Biomol Screen. 2003 Oct;8(5):566-70. doi: 10.1177/1087057103256465.
Several factors are known to increase the noise and variability of cell-based assays used for high-throughput screening. In particular, edge effects can result in an unacceptably high plate rejection rate in screening runs. In an effort to minimize these variations, the authors analyzed a number of factors that could contribute to edge effects in cell-based assays. They found that pre-incubation of newly seeded plates in ambient conditions (air at room temperature) resulted in even distribution of the cells in each well. In contrast, when newly seeded plates were placed directly in the CO(2) incubator, an uneven distribution of cells occurred in wells around the plate periphery, resulting in increased edge effect. Here, the authors show that the simple, inexpensive approach of incubating newly seeded plates at room temperature before placing them in a 37 degrees C CO(2) incubator yields a significant reduction in edge effect.
已知有几个因素会增加用于高通量筛选的基于细胞的检测的噪音和变异性。特别是,边缘效应可能导致筛选过程中平板拒收率高得令人无法接受。为了尽量减少这些变化,作者分析了一些可能导致基于细胞的检测中出现边缘效应的因素。他们发现,将新接种的平板在环境条件(室温空气)下预孵育会导致每个孔中的细胞分布均匀。相比之下,当新接种的平板直接放入二氧化碳培养箱中时,平板周边孔中的细胞会出现不均匀分布,从而导致边缘效应增加。在此,作者表明,在将新接种的平板放入37摄氏度的二氧化碳培养箱之前,先在室温下孵育这种简单且成本低廉的方法可显著降低边缘效应。