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Updated: Mar 24, 2026
Imaging and 3D Reconstruction of Cerebrovascular Structures in Embryonic Zebrafish
Published on: April 22, 2014
Zebrafish Caudal Fin Angiogenesis Assay-Advanced Quantitative Assessment Including 3-Way Correlative Microscopy
Ruslan Hlushchuk1, Daniel Brönnimann1, Carlos Correa Shokiche1
1Institute of Anatomy, University of Bern, Bern, Switzerland.
Background:
Researchers evaluating angiomodulating compounds as a part of scientific projects or pre-clinical studies are often confronted with limitations of applied animal models. The rough and insufficient early-stage compound assessment without reliable quantification of the vascular response counts, at least partially, to the low transition rate to clinics.
Objective:
To establish an advanced, rapid and cost-effective angiogenesis assay for the precise and sensitive assessment of angiomodulating compounds using zebrafish caudal fin regeneration. It should provide information regarding the angiogenic mechanisms involved and should include qualitative and quantitative data of drug effects in a non-biased and time-efficient way.
Approach & Results:
Basic vascular parameters (total regenerated area, vascular projection area, contour length, vessel area density) were extracted from in vivo fluorescence microscopy images using a stereological approach. Skeletonization of the vasculature by our custom-made software Skelios provided additional parameters including "graph energy" and "distance to farthest node". The latter gave important insights into the complexity, connectivity and maturation status of the regenerating vascular network. The employment of a reference point (vascular parameters prior amputation) is unique for the model and crucial for a proper assessment. Additionally, the assay provides exceptional possibilities for correlative microscopy by combining in vivo-imaging and morphological investigation of the area of interest. The 3-way correlative microscopy links the dynamic changes in vivo with their structural substrate at the subcellular level.
Conclusions:
The improved zebrafish fin regeneration model with advanced quantitative analysis and optional 3-way correlative morphology is a promising in vivo angiogenesis assay, well-suitable for basic research and preclinical investigations.
Insights
This study presents an improved zebrafish fin regeneration model for assessing blood vessel formation. The assay offers precise, quantitative analysis of angiomodulating compounds, aiding drug development.
Area of Science:
- Vascular biology
- Regenerative medicine
- Zebrafish models
Background:
- Current animal models for evaluating angiomodulating compounds have limitations.
- Insufficient early-stage assessment hinders clinical translation of vascular drugs.
Purpose of the Study:
- To develop a rapid, cost-effective angiogenesis assay using zebrafish caudal fin regeneration.
- To precisely assess angiomodulating compounds, providing mechanistic and quantitative data.
Main Methods:
- Utilized in vivo fluorescence microscopy and stereology for vascular parameter extraction.
- Employed custom software (Skelios) for vascular skeletonization and analysis.
- Integrated correlative microscopy for in vivo and subcellular structural analysis.
Main Results:
- Extracted key vascular parameters including regenerated area and vessel density.
- Quantified vascular network complexity and maturation using 'graph energy' and 'distance to farthest node'.
- Established a unique reference point for accurate pre- and post-amputation comparison.
Conclusions:
- The enhanced zebrafish model provides a robust in vivo angiogenesis assay.
- Suitable for basic research and preclinical evaluation of angiomodulating compounds.
- Offers advanced quantitative analysis and correlative morphology for comprehensive drug assessment.
