Wu, Yicong et al. (2011) International Worm Meeting "A selective plane illumination microscope for high-speed, long-term C. elegans embryogenesis studies."

Rondeau, Gary, Shroff, Hari, Colon-Ramos, Daniel, Santella, Anthony, Bao, Zhirong, Christensen, Ryan, Ghitani, Alireza, Wu, Yicong

[International Worm Meeting, 2011]

We introduce a high speed selective plane illumination microscope (SPIM) for in toto studies of development in C. elegans. The system allows continuous visualization of fluorescent protein constructs in transgenic nematode embryos, from fertilization through hatching with no detectable photodamage. Volumetric images are collected every two seconds, for a total of ~24,000 imaging volumes over the entire 14 hours of embryogenesis. The high imaging rate afforded by the microscope minimizes artifacts due to motion blur and enables examination of developmental events through twitching without the use of anesthetics or genetic perturbations. The microscope is 30 times faster than spinning disk confocal microscopy, but the signal-to-noise ratio is superior and subcellular resolution is comparable, allowing visualization of cell biological events throughout development. We have combined the system with existing computer-assisted cell identification approaches to generate a module that enables unambiguous identification of individual cells through the use of lineage identity. Our results establish a strategy and pave the way for systematic mapping of the wiring process and extension of the wiring diagram to a four dimensional dynamic atlas. The microscope design employs a straightforward add-on of a SPIM module onto an inverted microscope and uses conventional mounting of specimens on glass slides, making it readily adaptable by other cell- and developmental biology labs.