Program in Systems Biology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
STAR Protoc. 2021 Feb 3;2(1):100327. doi: 10.1016/j.xpro.2021.100327. eCollection 2021 Mar 19.
Evaluating drug sensitivity is improved by directly quantifying death kinetics, rather than correlates of viability, such as metabolic activity. This is challenging, requiring time-lapse microscopy and genetically encoded labels to distinguish live and dead cells. Here, we describe fluorescence-based and lysis-dependent inference of cell death kinetics (FLICK). This method requires only a standard fluorescence plate reader, retaining the high-throughput nature and broad accessibility of common viability assays. However, FLICK specifically quantifies death, including an accurate inference of death kinetics. For complete details on the use and execution of this protocol, please refer to Richards et al. (2020).
通过直接量化死亡动力学，而不是代谢活性等生存能力相关指标，来提高药物敏感性的评估。这是一项具有挑战性的任务，需要使用延时显微镜和遗传编码标签来区分活细胞和死细胞。在这里，我们描述了基于荧光和基于裂解的细胞死亡动力学推断（FLICK）。这种方法仅需要标准的荧光板读数器，保留了常见生存力测定的高通量性质和广泛的可及性。然而，FLICK 专门定量死亡，包括对死亡动力学的准确推断。有关该协议的使用和执行的完整详细信息，请参阅 Richards 等人。（2020 年）。
