Movie caption: A five-hour time-lapse of dendritic branching in a C. elegans PVD neuron, acquired on diSPIM light sheet microscope. The video captures dendritic branching on the dorsal and ventral sides of the midbody section. Credit: Jayson Smith et al., Frontiers in Cell and Developmental Biology, 2022.
“The innovation we describe is essentially a combination of two known mounting approaches,” Wolff says. “One is a biopolymer that is viscous during sample preparation, but once you expose it to UV light it hardens and keeps the sample (in this case, C. elegans) immobile. The second part is a mounting method in plastic tubes that allows the use of light sheet microscopy. The combination allows one to live-image adult C. elegans over a period of more than 2 hours. It sounds like a short time period, but because of previous problems with immobilizing the specimen, this was not possible. Also, imaging from different angles, which LSFM allows, wasn’t possible before because of the specimen’s constant body movement.”
The team used the protocol to timelapse image a sensory neuron’s dendrites branching and pruning. And they expect it will enable better live-imaging studies of other important cell and developmental processes, such as germ stem cell biology, cell migration, cell division and cell invasion. The protocol is generalizable to work with other organisms, with little or no modifications.
“The paper demonstrates the privilege of being at with amazing people,” says Wolff, a co-author on the paper. The other authors are Embryology course teaching assistant Jayson J. Smith, a postdoc at University of Chicago; course teaching assistant Isabel Kenny, a doctoral candidate at Duke University; David Matus of Stony Brook University; course director David Sherwood of Duke University; Investigator and CZI Scientist Abhishek Kumar; and Research Assistant Rachel Cray.
Other participants in the Embryology course, Senior Aquarist Jonathan Henry, and Central Microscopy Facility staff are acknowledged in the paper for their assistance.
Citation:
Smith, Jayson et al. (2022) A light sheet fluorescence microscopy protocol for Caenorhabditis elegans larvae and adults. Frontiers in Cell and Developmental Biology, DOI: