Independent Researcher, Sunnyvale, CA 94089, USA.
Oxidative Stress Group, Department of Molecular Biosciences, University of South Florida, Tampa, FL 33620, USA.
Int J Mol Sci. 2023 Apr 13;24(8):7187. doi: 10.3390/ijms24087187.
A comet assay is a trusted and widely used method for assessing DNA damage in individual eukaryotic cells. However, it is time-consuming and requires extensive monitoring and sample manipulation by the user. This limits the throughput of the assay, increases the risk of errors, and contributes to intra- and inter-laboratory variability. Here, we describe the development of a device which automates high throughput sample processing for a comet assay. This device is based upon our patented, high throughput, vertical comet assay electrophoresis tank, and incorporates our novel, patented combination of assay fluidics, temperature control, and a sliding electrophoresis tank to facilitate sample loading and removal. Additionally, we demonstrated that the automated device performs at least as well as our "manual" high throughput system, but with all the advantages of a fully "walkaway" device, such as a decreased need for human involvement and a decreased assay run time. Our automated device represents a valuable, high throughput approach for reliably assessing DNA damage with the minimal operator involvement, particularly if combined with the automated analysis of comets.
彗星分析是一种用于评估单个真核细胞 DNA 损伤的可靠且广泛使用的方法。然而,它耗时且需要用户进行广泛的监测和样本操作。这限制了测定的通量,增加了出错的风险,并导致了实验室内部和实验室之间的可变性。在这里,我们描述了一种自动化高通量样本处理彗星分析的设备的开发。该设备基于我们的专利、高通量、垂直彗星分析电泳槽,并结合了我们新颖的、专利的组合分析流体、温度控制和滑动电泳槽,以方便样本加载和去除。此外,我们证明自动化设备的性能至少与我们的“手动”高通量系统一样好,但具有全“免提”设备的所有优点,例如减少对人员的参与和减少测定运行时间的需求。我们的自动化设备代表了一种有价值的高通量方法,可在最小的操作人员参与下可靠地评估 DNA 损伤,特别是如果与彗星的自动分析相结合。