Capturing tissue repair in zebrafish larvae with time-lapse brightfield stereomicroscopy.
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.
Davis Center for Regenerative Biology and Medicine, MDI Biological Laboratory.
Thomas S
TS
Lisse
Davis Center for Regenerative Biology and Medicine, MDI Biological Laboratory.
Elizabeth A
EA
Brochu
Davis Center for Regenerative Biology and Medicine, MDI Biological Laboratory; srieger@mdibl.org.
Sandra
S
Rieger
D000818:Animals; D008853:Microscopy / Q000379:methods; D012038:Regeneration / Q000502:physiology*; D013623:Tail / Q000502:physiology; D015027:Zebrafish / Q000502:physiology*