FIGURE SUMMARY
Title

Protocol development for discovery of angiogenesis inhibitors via automated methods using zebrafish

Authors
Mauro, A., Ng, R., Li, J.Y., Guan, R., Wang, Y., Singh, K.K., Wen, X.Y.
Source
Full text @ PLoS One
Fig 1

Top view of a 3D CAD drawing of the HTP.

A: Sciclone G3 Advanced Liquid Handler. B: Twister II robots with transfer stations between them. C: T-Robot thermocycler. D: Multidrop 384. E: ELx405 plate washer. F: Synergy H1 plate reader. G: Image Xpress Ultra confocal microscope. H: COPAS XL embryo sorter. I: STX220 Incubator. J: LPX220 plate hotel. Scale bar is 50 cm.
Fig 2

Flow chart of the HTP drug screening protocol for endothelial cell modulators.

Grey font: time of plate movement. Red font: contents of plate while in transit between devices. Blue font: action taken at device.
Fig 3

Visualization of the pixel count readout.

The image on the left is a composite of 2D images from each of the wells of a 96-well plate as presented by MetaXpress. The images on the right are blown up unprocessed and processed images from a well with 0.05% DMSO (negative control) and a well with 8 μM I3M (angiogenesis inhibitor). Scale bar is 700 μm.
Fig 4

Pixel count readout results that validate the high throughput protocol developed for the discovery of angiogenesis inhibitors.

Tg(kdrl:EGFP) zebrafish were treated with 0.05% DMSO and compared to Tg(kdrl:EGFP) zebrafish treated with various concentrations of I3M to validate protocol. The experiment was repeated in triplicate. The error bars represent the standard deviation of the means from the three experiments.
Fig 5

Survival curve for Tg(kdrl:EGFP) zebrafish treated with various concentrations of I3M.

The experiment was repeated in triplicate. The error bars represent the standard deviation of the survival percentage from the three experiments.
Fig 6

Optimized COPAS pressure settings to dispense 1 embryo/well.

The “Never stop dispensing” box is checked so that dispensing does not stop due to the assigned pressures falling out of the default range.
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ PLoS One
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