Capturing tissue repair in zebrafish larvae with time-lapse brightfield stereomicroscopy

Journal of Visualized Experiments : JoVE
Thomas S LisseSandra Rieger

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.

Citations

Apr 2, 2016·Proceedings of the National Academy of Sciences of the United States of America·Thomas S LisseSandra Rieger
Jan 1, 2019·Bio-protocol·Zhiqiang ZengE Elizabeth Patton
Apr 8, 2020·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Rongrong ZhaoChusen Huang

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Methods Mentioned

BETA
confocal microscopy
imaging techniques
amputation
scraping
Assay
amputations
imaging procedure

Software Mentioned

Axiovision
ImageJ
Fiji
Imaris
StackReg
Deconvolve
Autoquant X3
Image J

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