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Pharmacology JoVE (Open Access) Citable · DOI

High-throughput Measurement of Gut Transit Time Using Larval Zebrafish

DOI: 10.3791/58497-v
What you'll learn
  • ✓ Train larval zebrafish to feed on fluorescently labeled food
  • ✓ Measure gut transit time using high-throughput fluorescence imaging
  • ✓ Quantify treatment effects on gastrointestinal motility in larvae
Protocol

The goal of this protocol is to measure the transit time of fluorescently labeled food through the gut of larval zebrafish in a high throughput fashion.

Difficulty
intermediate
Total time
~3–4 hours (including larval acclimation, feeding, and imaging)
Model organism
Danio rerio (larval zebrafish)
Biosafety
BSL-1

Steps

1
Train larvae to feed using non-dyed food

Acclimate larval zebrafish to feeding conditions by providing non-fluorescent food to establish baseline feeding behavior. This step ensures larvae are ready to consume the fluorescently labeled food in subsequent steps.

▶ 01:27
2
Prepare fluorescent food and feed larvae

Generate fluorescently labeled food, administer it to trained larvae, and apply experimental treatments. This establishes the baseline timepoint for tracking food movement through the gut.

▶ 02:10
3
Measure initial and subsequent fluorescence

Quantify fluorescence intensity in larval wells at multiple timepoints using a plate reader or imaging system. Sequential measurements track the progression of labeled food through the gastrointestinal tract.

▶ 04:08
4
Analyze high-throughput gut transit time data

Calculate transit time kinetics from fluorescence measurements to determine how quickly labeled food moves through the larval gut under different conditions. Results enable assessment of treatment effects on gastrointestinal motility.

▶ 05:04
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