Department of Molecular Life Science, University Zürich, Winterthurerstrasse 190, 8057 Zürich, Switzerland.
Institute for Chemical and Bioengineering, ETH Zürich, 8093 Zürich, Switzerland.
Development. 2021 Jul 15;148(18). doi: 10.1242/dev.199674. Epub 2021 Jul 21.
Several microfluidic-based methods for Caenorhabditis 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 that 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.
最近已经提出了几种基于微流控的秀丽隐杆线虫成像方法。现有的方法要么允许在不保持稳定的线虫方向的情况下对多个幼虫阶段进行成像，要么允许非常好的固定化，但只适用于较短的实验。在这里，我们提出了一种新颖的微流控成像方法，该方法允许在多个幼虫阶段进行平行活体成像，同时随着时间的推移保持线虫的方向和身份。这是通过精心调整的微流道阵列实现的，该阵列能够将线虫保持在稳定的方向，同时允许生长和蜕皮发生。通过一个主动的液压阀实现固定化，该阀仅在获取图像时将线虫压在盖玻片上。通过这种方式，可以以最小的对线虫活力或发育时间的影响获得高质量的图像。该设备的功能通过观察皮下 seam 和 P 细胞分裂来证明，并且首次观察到从诱导到形态发生结束的整个过程。此外，我们通过在锚定细胞入侵过程中干扰基膜突破来证明芯片上 RNAi 的可行性。
