title: Zebrafish Caudal Fin Angiogenesis Assay-Advanced Quantitative Assessment Including 3-Way Correlative Microscopy.
abstract: 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.
  
  
  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.
authors.affiliation: Institute of Anatomy, University of Bern, Bern, Switzerland.
authors.forename: Ruslan
authors.initials: R
authors.lastname: Hlushchuk
authors.affiliation: Institute of Anatomy, University of Bern, Bern, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Brönnimann
authors.affiliation: Institute of Anatomy, University of Bern, Bern, Switzerland.
authors.forename: Carlos
authors.initials: C
authors.lastname: Correa Shokiche
authors.affiliation: Institute of Anatomy, University of Bern, Bern, Switzerland.
authors.forename: Laura
authors.initials: L
authors.lastname: Schaad
authors.affiliation: Institute of Anatomy, University of Bern, Bern, Switzerland.
authors.forename: Ramona
authors.initials: R
authors.lastname: Triet
authors.affiliation: Department of Biology, University of Fribourg, Fribourg, Switzerland.
authors.forename: Anna
authors.initials: A
authors.lastname: Jazwinska
authors.affiliation: Institute of Anatomy, University of Bern, Bern, Switzerland.
authors.forename: Stefan A
authors.initials: SA
authors.lastname: Tschanz
authors.affiliation: Institute of Anatomy, University of Bern, Bern, Switzerland.
authors.forename: Valentin
authors.initials: V
authors.lastname: Djonov
mesh_terms: D058500:Animal Fins / Q000098:blood supply* / Q000187:drug effects / Q000502:physiology; D000818:Animals; D003362:Cost-Benefit Analysis; D008853:Microscopy / Q000191:economics / Q000379:methods*; D018919:Neovascularization, Physiologic / Q000187:drug effects; D010793:Phthalazines / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D012038:Regeneration / Q000187:drug effects; D013623:Tail; D013997:Time Factors; D015027:Zebrafish
chemical_list: D010793:Phthalazines; D011725:Pyridines; C404768:vatalanib