The automated analysis of angiogenesis in Zebrafish using ArrayScan acquisition and Definiens image analysis. Abrogation of angiogenesis was achieved by blocking the expression of the VEGF receptor using a morpholino oligonucleotide targeting mRNA translation. The morpholino was injected directly into the oocyte before it reached the 8 cell stage. Untreated (panels A & B) and treated (panel C) fish images were captured at day 4. The Definiens algorithm first identifies the fish (in yellow) then the yolk sack (brown region) and from the yolk sack position creates a dorsal region of interest (white area). The central blood vessel inside this region is then found (red region). Finally, the individual intersegmental vessels (ISV) are delineated in such a way that each individual ISV is treated as a separate object allowing detailed statistical analysis on each individual vessel. The number of ISV blood vessels are significantly decreased in the morpholino treated animals (panel D) Likewise the ISVs and central blood vessel area is decreased along with a concomitant decrease in the dorsal area (panel E) showing that the overall development of the dorsal area is highly dependent on blood vessel supply. Each data point is the meanąSEM of 41 control and 15 morpholino treated fish respectively.

Zebrafish

With the use of more advanced image analysis software, it is possible to extend HCA beyond the analysis of simple cells. With the help of a grant from the Northwest Development agency, we have been developing automated zebrafish analysis. Our aim is to use these creatures to gain rich phenotypic signatures on which we can cluster compounds commonly used in the clinic. These signatures will then be used to explore compound reprofiling.