| Title | MiniSOG-mediated Photoablation in Caenorhabdtis elegans |
| Author | Yingchuan Qi, Suhong Xu |
| Affiliation(s) | Division of Biological Sciences, Neurobiology Section, University of California, San Diego, CA, USA |
| Published | bio-protocol Vol 3, Iss 1, January 05, 2013 DOI: 10.21769/BioProtoc.316
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| Keyword | UHP-Mic-LED-460 |
| Snippet | |
| Abstract | Abstract
This protocol describes a method for light-inducible cell ablation in live worms. miniSOG (mini Singlet Oxygen Generator) generates singlet oxygen upon blue light illumination (Shu et al., 2011). Mitochondrially membrane targeted miniSOG (the first 55 a. a. of C. e. tomm-20 fused at the N’-terminus of miniSOG, termed as mito-miniSOG in the following) is transgenically expressed in specific cells/tissues (Qi et al., 2012). Groups of transgenic animals are illuminated under open field fluorescence light on a compound microscope or LED light setup for photo-ablation.
Equipment
Fluorescence stereomicroscope (dissecting microscope, e.g., Leica, model: M165 )
Upright epi-fluorescecne microscope (compound microscope, e.g., Zeiss Axio Imager 2)
Prizmatix LED (UHP-Mic-LED-460), and Digital Function Generator (PASCO Scientific, catalog number: PI-9587C )
Microinjection rig to generate transgenic worms |
