A016 - Parallel long-term live imaging of developing C. elegans larvae with microfluidics

Wolfgang Keil (wolfgang.keil@curie.fr)
Eliot Schlang (eliot.schlang@curie.fr)

Long-term live imaging of developing multicellular organisms is mostly performed in embryos, which remain in a fixed position and do not grow substantially during image acquisition. Imaging post-embryonic stages in intact animals is incomparably more difficult because samples are moving, feeding, and growing while development takes place. The objective of the workshop is to introduce participants to the model organism C. elegans and a simple yet powerful microfluidic system to image its post-embryonic development that we now routinely use in our lab (Berger et al. Development 2021). In this system, C. elegans larva are confined in small micro-channels. Channels are periodically constricted with pressure to limit movement and allow high-resolution volumetric imaging. The microfluidics device enables imaging up to 40 developing animals in parallel for periods of up to 2 days to capture cell-divisions and gene expression time courses at single cell resolution throughout the animal body. In the workshop, we will setup a microfluidics experiment with a strain in which skin stem cells and their descendants are labelled with mCherry and a key stem-cell fate-determining transcription factor is CRISPR-tagged with split-GFP. We will observe and track stem cell divisions as well as the evolution of split-GFP intensity over time in the individual cell lineages. A data set obtained in our lab with an identical strain will be used to introduce participants to post-acquisition analysis, focusing on challenges specific to moving samples, low-intensity fluorescent reporters and large 5D imaging datasets.