The Photoconversion Technique for Exploring Inflammatory Cell Dynamics in Insect Pupae
DEŞİFRE METNİ
Begin with a glass-bottomed dish containing transgenic Drosophila pupae carrying labeled wounded epithelium with their cell surface proteins exhibiting green fluorescence.
The pupae's immune cells — or hemocytes express photoconvertible green fluorophores in the cytoplasm and red fluorescent proteins in the nucleus, facilitating cell tracking.
The wounded cells release chemoattractants — or inflammatory molecules, attracting the hemocytes to the wounded site.
Confocal microscopy reveals the wounded area encircled by green epithelial cells.
During healing, green hemocytes with red nuclei near the wound extend membrane protrusions — filopodia and lamellipodia and migrate toward the wounded area.
Over time, as the chemoattractant diffuses, distant hemocytes migrate toward the wound, creating an immune cell wave.
Illuminate the subpopulation of migratory hemocytes with a 405-nanometer wavelength. This irreversibly converts the hemocytes' photoconvertible fluorophore from green to red.
These photoconverted hemocytes repair the wounded epithelium and move away from the wound site, differentiating the photoconverted hemocytes from non-photoconverted hemocytes, and elucidating inflammatory cell dynamics.
