The roundworm C. elegans is a mainstay of aging research due to its short lifespan and easily manipulable genetics. Current, widely used methods for long-term measurement of C. elegans are limited by low throughput and the difficulty of performing longitudinal monitoring of aging phenotypes. Here we describe the WorMotel, a microfabricated device for long-term cultivation and automated longitudinal imaging of large numbers of C. elegans confined to individual wells. Using the WorMotel, we find that short-lived and long-lived strains exhibit patterns of behavioral decline that do not temporally scale between individuals or populations, but rather resemble the shortest and longest lived individuals in a wild type population. We also find that behavioral trajectories of worms subject to oxidative stress resemble trajectories observed during aging. Our method is a powerful and scalable tool for analysis of C. elegans behavior and aging.DOI: http://dx.doi.org/10.7554/eLife.26652.001
Phenotypic assays are crucial in genetics; however, traditional methods that rely on human observation are unsuitable for quantitative, large-scale experiments. Furthermore, there is an increasing need for comprehensive analyses of multiple phenotypes to provide multidimensional information. Here we developed an automated, high-throughput computer imaging system for quantifying multiple Caenorhabditis elegans phenotypes. Our imaging system is composed of a microscope equipped with a digital camera and a motorized stage connected to a computer running the QuantWorm software package. Currently, the software package contains one data acquisition module and four image analysis programs: WormLifespan, WormLocomotion, WormLength, and WormEgg. The data acquisition module collects images and videos. The WormLifespan software counts the number of moving worms by using two time-lapse images; the WormLocomotion software computes the velocity of moving worms; the WormLength software measures worm body size; and the WormEgg software counts the number of eggs. To evaluate the performance of our software, we compared the results of our software with manual measurements. We then demonstrated the application of the QuantWorm software in a drug assay and a genetic assay. Overall, the QuantWorm software provided accurate measurements at a high speed. Software source code, executable programs, and sample images are available at www.quantworm.org. Our software package has several advantages over current imaging systems for C. elegans. It is an all-in-one package for quantifying multiple phenotypes. The QuantWorm software is written in Java and its source code is freely available, so it does not require use of commercial software or libraries. It can be run on multiple platforms and easily customized to cope with new methods and requirements.
Pine wilt disease caused by Bursaphelenchus xylophilus is a major tree disease that threatens pine forests worldwide. To diagnose this disease, we developed battery-powered remote sensing devices capable of long-range (LoRa) communication and installed them in pine trees (Pinus densiflora) in Gyeongju and Ulsan, South Korea. Upon analyzing the collected tree sensing signals, which represented stem resistance, we found that the mean absolute deviation (MAD) of the sensing signals was useful for distinguishing between uninfected and infected trees. The MAD of infected trees was greater than that of uninfected trees from August of the year, and in the two-dimensional plane, consisting of the MAD value in July and that in October, the infected and uninfected trees were separated by the first-order boundary line generated using linear discriminant analysis. It was also observed that wood moisture content and precipitation affected MAD. This is the first study to diagnose pine wilt disease using remote sensors attached to trees.
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