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Capturing Tissue Repair in Zebrafish Larvae with Time-lapse Brightfield Stereomicroscopy
Authors:Thomas S Lisse  Elizabeth A Brochu  Sandra Rieger
Institution:1Davis Center for Regenerative Biology and Medicine, MDI Biological Laboratory
Abstract:The zebrafish larval tail fin is ideal for studying tissue regeneration due to the simple architecture of the larval fin-fold, which comprises of two layers of skin that enclose undifferentiated mesenchyme, and because the larval tail fin regenerates rapidly within 2-3 days. Using this system, we demonstrate a method for capturing the repair dynamics of the amputated tail fin with time-lapse video brightfield stereomicroscopy. We demonstrate that fin amputation triggers a contraction of the amputation wound and extrusion of cells around the wound margin, leading to their subsequent clearance. Fin regeneration proceeds from proximal to distal direction after a short delay. In addition, developmental growth of the larva can be observed during all stages. The presented method provides an opportunity for observing and analyzing whole tissue-scale behaviors such as fin development and growth in a simple microscope setting, which is easily adaptable to any stereomicroscope with time-lapse capabilities.
Keywords:Developmental Biology  Issue 95  Fin regeneration  Time-lapse imaging  Stereomicroscope  Heated incubation chamber  Post-imaging analysis
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