![]() ![]() The hydrogel layer is clamped between a coverslip (blue, bottom) and a cover glass (blue, top). ![]() ( a) Schematic cross-section of a single microchamber. Hydrogel microchambers have been used to constrain nematode movement for studying behaviour 21, but so far not development. elegans culture on agar plates and the established microscopy protocols for studying nematode development 2. Instead, animals move and feed under conditions similar to standard C. Second, in contrast to microfluidics, microchambers do not require using liquid culture. Microchambers have two main advantages over active microfluidics: first, they are simple to use, requiring no moving parts or flow. Finally, we use image analysis to track the dynamics of cells inside the animal’s body. Next, we use fast image acquisition to capture sharp images of larvae as they move inside each microchamber, precluding the need for immobilization altogether. 1): first, we constrain larval movement to the field of view of the microscope using microfabricated hydrogel chambers containing bacteria as food. elegans post-embryonic development we instead use a different approach ( Fig. Experiments that did support normal larval growth so far lacked the resolution to study development at the single-cell level 18, 19, 20. Microfluidics has been used to immobilize nematodes for microscopy by mechanical clamping 11, 12, flow 13, 14 or changes in the physicochemical environment 15, 16, 17 however, most of these devices are geared towards immobilizing adult nematodes and are not designed to support sustained development. Immobilizing larvae either mechanically or by paralysis-inducing drugs allows time-lapse microscopy only for limited time periods, as it prevents the animal from feeding, resulting in developmental arrest within hours 9, 10. elegans larvae are highly motile and thus are difficult to image at high magnification. However, long-term time-lapse microscopy is currently rarely used to study C. elegans uniquely suited to study the interplay between development and environmental cues such as diet, food availability and pheromones 6, 7, 8. Owing to their small and transparent anatomy, nematodes such as Caenorhabditis elegans are currently the only animals in which the entire development from embryo to adult can in principle be studied with single-cell resolution 2, 3, 4, 5. However, in these model organisms time-lapse microscopy is typically restricted to early stages of embryonic development. Recent advances in microscopy have made it possible to follow the dynamics of many, if not all cells in the development of entire zebrafish and fruit fly embryos 1. ![]()
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