The Replica Set Method: A High-throughput Approach to Quantitatively Measure Caenorhabditis elegans Lifespan
The Replica Set Method: A High-throughput Approach to Quantitatively Measure Caenorhabditis elegans Lifespan
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DOI：
10.3791/57819
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发表时间：
2018-06-01
影响因子：
1.2
通讯作者：
Samuelson, Andrew, V
中科院分区：
文献类型：
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作者：
Cornwel, Adam B.;Llop, Jesse R.;Samuelson, Andrew, V
该方法是一种以高通量方式定量测量秀丽隐杆线虫的寿命或存活率的方法，从而允许单个研究者在相同的时间内筛选更多的治疗或条件，而不会损失数据质量。该方法需要在大多数实验室中使用C。优雅，因此易于采用。该方法的核心是在每个观察点分析一个群体的独立样本，而不是像传统的纵向方法那样随着时间的推移分析单个样本。评分需要向多孔板的威尔斯孔中加入液体，以刺激C。elegans移动和促进量化的变化健康跨度。该方法的其他主要优点包括减少琼脂表面暴露于空气传播的污染物（例如霉菌或真菌），最小化动物处理，以及对零星错误评分（例如当动物仍然活着时将其称为死亡）的鲁棒性。为了适当地分析和可视化来自一个CNOSSET风格实验的数据，还开发了一个定制的软件工具。该软件目前的功能包括绘制生存曲线的两个TISET和传统的（卡普兰-迈耶）实验，以及统计分析的TISET。这里提供的协议描述了传统的实验方法和数据集方法，以及相应的数据分析的概述。
The Replica Set method is an approach to quantitatively measure lifespan or survival of Caenorhabditis elegans nematodes in a high-throughput manner, thus allowing a single investigator to screen more treatments or conditions over the same amount of time without loss of data quality. The method requires common equipment found in most laboratories working with C. elegans and is thus simple to adopt. The approach centers on assaying independent samples of a population at each observation point, rather than a single sample over time as with traditional longitudinal methods. Scoring entails adding liquid to the wells of a multi-well plate, which stimulates C. elegans to move and facilitates quantifying changes in healthspan. Other major benefits of the Replica Set method include reduced exposure of agar surfaces to airborne contaminants (e.g. mold or fungus), minimal handling of animals, and robustness to sporadic mis-scoring (such as calling an animal as dead when it is still alive). To appropriately analyze and visualize the data from a Replica Set style experiment, a custom software tool was also developed. Current capabilities of the software include plotting of survival curves for both Replica Set and traditional (Kaplan-Meier) experiments, as well as statistical analysis for Replica Set. The protocols provided here describe the traditional experimental approach and the Replica Set method, as well as an overview of the corresponding data analysis.