Several microfluidic-based methods for C. elegans imaging have recently been introduced. Existing methods either permit imaging across multiple larval stages without maintaining a stable worm orientation, or allow for very good immobilization but are only suitable for shorter experiments. Here, we present a novel microfluidic imaging method, which allows parallel live-imaging across multiple larval stages, while maintaining worm orientation and identity over time. This is achieved through an array of microfluidic trap channels carefully tuned to maintain worms in a stable orientation, while allowing growth and molting to occur. Immobilization is supported by an active hydraulic valve, which presses worms onto the cover glass during image acquisition only. In this way, excellent quality images can be acquired with minimal impact on worm viability or developmental timing. The capabilities of the devices are demonstrated by observing the hypodermal seam and P cell divisions and, for the first time, the entire process of vulval development from induction to the end of morphogenesis. Moreover, we demonstrate feasibility of on-chip RNAi by perturbing basement membrane breaching during anchor cell invasion.
Microfluidic-based imaging of complete C. elegans larval development
Currently Viewing Accepted Manuscript - Newer Version Available
Simon Berger, Silvan Spiri, Andrew deMello, Alex Hajnal; Microfluidic-based imaging of complete C. elegans larval development. Development 2021; dev.199674. doi: https://doi.org/10.1242/dev.199674
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