| Outline of Final Research Achievements |
To observe the interaction between various organelles and the cytoskeleton in the growth cones that lead neurites, we applied super-resolution shadow imaging, SUSHI, to visualize the intracellular structure. Using cultured cells that cytosolically expressed the fluorescent protein Citrine, we could readily discriminate negative images of various organelles, such as mitochondria, endoplasmic reticulum, endosome, lysosome and also cytoskeleton. The live shadow imaging showed that vesicles arising following actin polymerization and organelles that repeatedly move to the leading edge against the actin retrograde flow. Mitochondrial fission was also detected immediately after a vesicle approached the mitochondrion in the growth cone. The super-resolution imaging is one of the best methods to visualize multiple organelles and cytoskeletons simultaneously.
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