Our lab has developed a new cooling stage and associated immobilization protocol for strong immobilization of entire C. elegans populations on their original cultivation plate. Counterintuitively, relatively warmer temperatures immobilize animals significantly more effectively than colder temperatures utilized in prior studies. This enhanced immobilization enables clear submicron-resolution fluorescence imaging, which is challenging to achieve with most current immobilization techniques. We demonstrate 64× magnification 3D imaging and timelapse recording of neurons in adults and embryos without motion blur. Compared to standard azide immobilization, cooling immobilization reduces the animal preparation and recovery time by >98%, significantly increasing experimental speed. By obviating individual animal manipulation, our approach could also empower automated imaging of large C. elegans populations within standard experimental setups and workflows.
We post the building manual here: #6
More information about the cooling stage building and using can be found here, and you are very welcomed to post your question or suggestion here: https://github.com/wormneurolab/Cooling-Stage/discussions
