A LED light for photo-inducible cell ablation by miniSOG

Photosensitizer MiniSOG (mini-singlet oxygen generator), a green fluorescent flavoprotein, can be used to kill cells in a spatially and temporally regulated manner according to a chosen promoter when it is fused with the target cells. However, it needs 470 nm blue light excitation to release reactive oxygen species (ROS) to work. In the past, researchers usually used fluorescence microscope to generate light, which wasted the lab resources and influenced other experiments. Here, we developed a homemade light emitting diode (LED) light for photo-inducible cell ablation by photosensitizers. The LED light consisted of six groups of lamps, and it could illuminate a plurality of culture dish drawers simultaneously with different light intensities. In order to watch the efficacy of cell ablation, confocal imaging, behavioral, calcium imaging and electrophysiology experiments have been performed on C. elegans. Most of neurons related with forward movements have been ablated after dozens of minutes light irradiation. The worms can only move backward with wriggled body, and the calcium oscillation increases in reverse movement motor neurons. The whole frequency and amplitude of miniature postsynaptic currents (PSCs) in body wall muscle decrease because of death of some neurons. These experimental results verified that the LED light was useful and convenient. This study provides a method for high-throughput cell ablation experiments.

Automated summary

Photosensitizer MiniSOG, a green fluorescent flavoprotein, can kill cells in a regulated manner with a specific light source. Researchers developed a homemade LED light for high-throughput cell ablation experiments.