Ecole Polytechnique Fédérale de Lausanne (EPFL), Laboratory of Microsystems, Lausanne, Switzerland.
Ecole Polytechnique Fédérale de Lausanne (EPFL), Laboratory of Integrative Systems Physiology, Lausa, Switzerland.
J Biomed Opt. 2018 Nov;24(2):1-9. doi: 10.1117/1.JBO.24.2.021205.
A major step for the validation of medical drugs is the screening on whole organisms, which gives the systemic information that is missing when using cellular models. Caenorhabditis elegans is a soil worm that catches the interest of researchers who study systemic physiopathology (e.g., metabolic and neurodegenerative diseases) because: (1) its large genetic homology with humans supports translational analysis; (2) worms are much easier to handle and grow in large amounts compared with rodents, for which (3) the costs and (4) the ethical concerns are substantial. Here, we demonstrate how multimodal optical imaging on such an organism can provide high-content information relevant to the drug development pipeline (e.g., mode-of-action identification, dose-response analysis), especially when combined with on-chip multiplexing capability. After designing a microfluidic array to select small separated populations of C. elegans, we combine fluorescence and bright-field imaging along with high-throughput feature recognition and signal detection to enable the identification of the mode-of-action of an antibiotic. For this purpose, we use a genetically encoded fluorescence reporter of mitochondrial stress, which we studied in living specimens during their entire development. Furthermore, we demonstrate real-time, very large field-of-view capability on multiplexed motility assays for the assessment of the dose-response relation of an anesthetic.
药物验证的一个主要步骤是在整个生物体上进行筛选，这可以提供使用细胞模型时所缺少的系统信息。秀丽隐杆线虫是一种土壤蠕虫，它引起了研究系统病理生理学（例如代谢和神经退行性疾病）的研究人员的兴趣，原因有三：（1）它与人类的遗传同源性很高，支持转化分析；（2）与啮齿动物相比，蠕虫更容易大量处理和生长，而（3）啮齿动物的成本和（4）伦理问题相当大。在这里，我们展示了如何对这种生物体进行多模态光学成像，以提供与药物开发管道相关的高内涵信息（例如，作用模式识别、剂量反应分析），尤其是与芯片上的多路复用能力相结合时。在设计了一种用于选择小而分离的秀丽隐杆线虫种群的微流控阵列之后，我们结合荧光和明场成像以及高通量特征识别和信号检测，以识别抗生素的作用模式。为此，我们使用了一种遗传编码的线粒体应激荧光报告基因，我们在活体标本的整个发育过程中对其进行了研究。此外，我们展示了用于评估麻醉剂剂量反应关系的实时、大视场能力的多路复用运动检测。
