School of Biological Sciences, University of Queensland, St. Lucia Campus, Brisbane, Queensland, Australia.
PLoS One. 2012;7(3):e33483. doi: 10.1371/journal.pone.0033483. Epub 2012 Mar 23.
There are four main phenotypes that are assessed in whole organism studies of Caenorhabditis elegans; mortality, movement, fecundity and size. Procedures have been developed that focus on the digital analysis of some, but not all of these phenotypes and may be limited by expense and limited throughput. We have developed WormScan, an automated image acquisition system that allows quantitative analysis of each of these four phenotypes on standard NGM plates seeded with E. coli. This system is very easy to implement and has the capacity to be used in high-throughput analysis.
METHODOLOGY/PRINCIPAL FINDINGS: Our system employs a readily available consumer grade flatbed scanner. The method uses light stimulus from the scanner rather than physical stimulus to induce movement. With two sequential scans it is possible to quantify the induced phototactic response. To demonstrate the utility of the method, we measured the phenotypic response of C. elegans to phosphine gas exposure. We found that stimulation of movement by the light of the scanner was equivalent to physical stimulation for the determination of mortality. WormScan also provided a quantitative assessment of health for the survivors. Habituation from light stimulation of continuous scans was similar to habituation caused by physical stimulus.
CONCLUSIONS/SIGNIFICANCE: There are existing systems for the automated phenotypic data collection of C. elegans. The specific advantages of our method over existing systems are high-throughput assessment of a greater range of phenotypic endpoints including determination of mortality and quantification of the mobility of survivors. Our system is also inexpensive and very easy to implement. Even though we have focused on demonstrating the usefulness of WormScan in toxicology, it can be used in a wide range of additional C. elegans studies including lifespan determination, development, pathology and behavior. Moreover, we have even adapted the method to study other species of similar dimensions.
在秀丽隐杆线虫的整体生物研究中，有四种主要表型进行评估：死亡率、运动、繁殖力和体型。已经开发出一些专注于这些表型的数字分析的程序，但这些程序可能受到费用和有限的通量的限制。我们开发了 WormScan，这是一种自动图像采集系统，可以对标准 NGM 板上接种的大肠杆菌进行这四种表型的定量分析。该系统非常易于实施，并且具有在高通量分析中使用的能力。
方法/主要发现：我们的系统采用现成的消费级平板扫描仪。该方法使用来自扫描仪的光刺激而不是物理刺激来诱导运动。通过两次连续扫描，可以量化诱导的趋光反应。为了证明该方法的实用性，我们测量了秀丽隐杆线虫对磷化氢气体暴露的表型反应。我们发现，通过扫描仪的光刺激来刺激运动与物理刺激一样可以用于确定死亡率。WormScan 还为幸存者提供了健康的定量评估。连续扫描的光刺激引起的习惯化与物理刺激引起的习惯化相似。
结论/意义：已经有用于秀丽隐杆线虫自动表型数据收集的现有系统。与现有系统相比，我们的方法具有以下特定优势：能够高通量评估更广泛的表型终点，包括死亡率的确定和幸存者的运动能力的量化。我们的系统还具有成本低廉和易于实施的特点。尽管我们专注于展示 WormScan 在毒理学中的有用性，但它可以用于广泛的其他秀丽隐杆线虫研究，包括寿命测定、发育、病理学和行为。此外，我们甚至已经改编了该方法来研究其他类似尺寸的物种。
