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Neuroscience JoVE (Open Access) Citable · DOI

Automated High-throughput Behavioral Analyses in Zebrafish Larvae

DOI: 10.3791/50622-v
What you'll learn
  • ✓ Set up automated high-throughput imaging system for zebrafish larval behavior detection
  • ✓ Collect and raise zebrafish embryos to 7-day-old larvae
  • ✓ Quantify behavioral responses to toxicants and pharmaceuticals using image analysis
Protocol

Our laboratory developed a novel high-throughput automated imaging system that is useful for the detection of several different behaviors in 7-day-old zebrafish larvae. The system can be used for detecting subtle changes in behavior after the larvae have been exposed to environmental toxicants or pharmaceuticals.

Difficulty
intermediate
Total time
~7–10 days (embryo collection through larval assay and analysis)
Model organism
Danio rerio (zebrafish larvae, 7 days post-fertilization)
Biosafety
BSL-1

Steps

1
Collect zebrafish embryos and culture larvae

Obtain zebrafish embryos from breeding tanks and raise them to 7 days post-fertilization under standard aquaculture conditions. Maintain appropriate water quality and feeding schedules.

▶ 01:48
2
Prepare molds for behavioral assay wells

Fabricate or assemble specialized molds containing wells designed to contain individual larvae during imaging. Ensure mold dimensions accommodate 7-day-old larvae.

▶ 03:08
3
Assemble and calibrate imaging cabinet system

Configure the automated imaging cabinet with appropriate lighting, temperature control, and camera positioning for consistent high-throughput video capture of larval behavior.

▶ 04:02
4
Capture behavioral video images of larvae

Position zebrafish larvae in prepared molds within the imaging cabinet and acquire automated video recordings of spontaneous and stimulus-evoked behaviors.

▶ 04:39
5
Analyze behavioral parameters from recorded images

Process video data using automated image analysis software to extract and quantify multiple behavioral metrics from each larva.

▶ 06:08
6
Interpret quantified behaviors and treatment effects

Examine behavioral outputs to identify subtle changes in larval responses following exposure to environmental toxicants or pharmaceutical agents.

▶ 07:31
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