FIGURE 1

FIGURE 1

In vivo dopamine neuron imaging-based high throughput screening in larval zebrafish identifies potential neuroprotective compounds. (A) Overview of the high throughput screening assay. 3 dpf larvae are transferred to 96 well plates with 10 μM screening compounds. DMSO (positive control) or 4.5 mM MTZ (negative control) was added 3 hours later and the treatment lasted for 24 hr, followed by imaging with brightfield and TexRed channels on InCell 6,000. Images were analyzed with the Cellprofiler pipeline. (B) Schematic of the image processing pipeline using the custom generated MATLAB “fishplatebrowser” and Cellprofiler. The brightfield and TexRed images were used to automatically detect the eye and diencephalic region of the brain and to quantify DA neurons. (C) Dual flashlight plot generated from custom made GUI “HitDataBrowser” with MATLAB. Compounds can be selected and exported with SSMD, BHS, and corresponding sample number. (D) Compounds in the top right quadrant with high BHS and SSMD scores based on manual selection. Details of the compounds are shown in Table 1. PTU: 1-phenyl 2-thiourea MTZ: Metronidazole DMSO: dimethyl sulfoxide.