Lochead Julia, Schessner Julia, Werner Tobias, Wölfl Stefan
Institute for Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany; Institute for Analytical Chemistry, Mannheim University of Applied Sciences, Mannheim, Germany.
Institute for Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany.
PLoS One. 2015 Jun 25;10(6):e0131233. doi: 10.1371/journal.pone.0131233. eCollection 2015.
Time-resolved cell culture assays circumvent the need to set arbitrary end-points and reveal the dynamics of quality controlled experiments. However, they lead to the generation of large data sets, which can represent a complexity barrier to their use. We therefore developed the Time-Resolved Cell Culture Assay (TReCCA) Analyser program to perform standard cell assay analyses efficiently and make sophisticated in-depth analyses easily available. The functions of the program include data normalising and averaging, as well as smoothing and slope calculation, pin-pointing exact change time points. A time-resolved IC50/EC50 calculation provides a better understanding of drug toxicity over time and a more accurate drug to drug comparison. Finally the logarithmic sensor recalibration function, for sensors with an exponential calibration curve, homogenises the sensor output and enables the detection of low-scale changes. To illustrate the capabilities of the TReCCA Analyser, we performed on-line monitoring of dissolved oxygen in the culture media of the breast cancer cell line MCF-7 treated with different concentrations of the anti-cancer drug Cisplatin. The TReCCA Analyser is freely available at www.uni-heidelberg.de/fakultaeten/biowissenschaften/ipmb/biologie/woelfl/Research.html. By introducing the program, we hope to encourage more systematic use of time-resolved assays and lead researchers to fully exploit their data.
时间分辨细胞培养分析方法避免了设置任意终点的需要,并揭示了质量控制实验的动态过程。然而,它们会产生大量数据集,这可能成为其应用的复杂性障碍。因此,我们开发了时间分辨细胞培养分析(TReCCA)分析仪程序,以高效地进行标准细胞分析,并轻松实现复杂的深入分析。该程序的功能包括数据归一化和平均化,以及平滑处理和斜率计算,可精确确定变化时间点。时间分辨IC50/EC50计算能更好地了解药物毒性随时间的变化情况,并实现更准确的药物间比较。最后,针对具有指数校准曲线的传感器的对数传感器重新校准功能,可使传感器输出均匀化,并能检测低水平变化。为了说明TReCCA分析仪的功能,我们对用不同浓度抗癌药物顺铂处理的乳腺癌细胞系MCF-7培养基中的溶解氧进行了在线监测。TReCCA分析仪可在www.uni-heidelberg.de/fakultaeten/biowissenschaften/ipmb/biologie/woelfl/Research.html免费获取。通过介绍该程序,我们希望鼓励更系统地使用时间分辨分析方法,并引导研究人员充分利用其数据。