Max Planck Research Group Neural Information Flow, Max Planck Institute for Neurobiology of Behavior - caesar, Bonn, Germany.
Institute of Medical Genetics, University of Zurich, Zurich, Switzerland.
Elife. 2022 Sep 9;11:e77252. doi: 10.7554/eLife.77252.
feeds on bacteria and other small microorganisms which it ingests using its pharynx, a neuromuscular pump. Currently, measuring feeding behavior requires tracking a single animal, indirectly estimating food intake from population-level metrics, or using restrained animals. To enable large throughput feeding measurements of unrestrained, crawling worms on agarose plates at a single worm resolution, we developed an imaging protocol and a complementary image analysis tool called PharaGlow. We image up to 50 unrestrained crawling worms simultaneously and extract locomotion and feeding behaviors. We demonstrate the tool's robustness and high-throughput capabilities by measuring feeding in different use-case scenarios, such as through development, with genetic and chemical perturbations that result in faster and slower pumping, and in the presence or absence of food. Finally, we demonstrate that our tool is capable of long-term imaging by showing behavioral dynamics of mating animals and worms with different genetic backgrounds. The low-resolution fluorescence microscopes required are readily available in laboratories, and in combination with our python-based analysis workflow makes this methodology easily accessible. PharaGlow therefore enables the observation and analysis of the temporal dynamics of feeding and locomotory behaviors with high-throughput and precision in a user-friendly system.
它通过咽部，即一个神经肌肉泵，摄取细菌和其他微生物作为食物。目前，测量进食行为需要跟踪单个动物，从种群水平的指标间接估计食物摄入量，或使用受约束的动物。为了能够在单个蠕虫分辨率下对琼脂平板上不受约束的爬行蠕虫进行高通量的进食测量，我们开发了一种成像方案和一种称为 PharaGlow 的互补图像分析工具。我们可以同时对多达 50 只不受约束的爬行蠕虫进行成像，并提取运动和进食行为。我们通过在不同的用例场景中测量进食，例如在发育过程中，通过遗传和化学干扰，导致泵送速度更快和更慢，以及在有或没有食物的情况下，证明了该工具的稳健性和高通量能力。最后，我们通过展示交配动物和具有不同遗传背景的蠕虫的行为动力学，证明了我们的工具能够进行长期成像。所需的低分辨率荧光显微镜在实验室中很容易获得，并且与我们基于 Python 的分析工作流程相结合，使这种方法易于使用。因此，PharaGlow 能够在用户友好的系统中以高通量和高精度观察和分析进食和运动行为的时间动态。
