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Method Article

Prey Capture Assay: A Method to Study the Prey Capture Behavior of Zebrafish Larva

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April 30th, 2023

In This Article

Abstract

Source: Muto, A., et al. Ablation of a Neuronal Population Using a Two-photon Laser and Its Assessment Using Calcium Imaging and Behavioral Recording in Zebrafish Larvae. J. Vis. Exp. (2018)

This video describes the prey capture assay. The video discusses the method to study prey capture behavior of zebrafish larvae after the laser ablation of specific neurons.

Protocol

1. Assessment of Behavioral Consequences Following Laser Ablation

  1. Set up a behavioral recording system that consists of a stereoscope equipped with a video camera. Use a camera with an appropriate image acquisition software, commercial or custom-made (Figure 1A).
  2. Optimize the lighting condition so that the paramecium can be detected by image processing. For example, use a ring white LED light with a spacer (Figure 1B).
    Note: The distance from, and the angle of, the LED affects the appearance of the sample (i.e., bright in dark background or dark in bright background) and ....

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Results

Stereoscope setup for image acquisition; microscopy experiment with lighting (ring LED) and data analysis.
Figure 1. Prey capture assay and representative data in laser-ablated zebrafish larvae. (A) Behavioral recording system. The stereomicroscope was equipped with a CMOS camera. Images were acquired using a custom-made script. (B) Compon.......

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Imageing ChambersGrace Bio-LabsCoverWell Imaging Chambers PCIA-2.5Used as a behavioral recording chamber
ORCA-Flash4.0 Hamamatsu PhotonicsModel:C11440-22CU A scientific CMOS camera
SZX7OlympusStereoscope
A ring LED lightCCS  Model: LDR2-100SW2-LAWhite LED
Secure-Seal Hybridization Chamber Gasket, 8 chambers, 9 mm diameter x 0.8 mm depthMolecular ProbesCatalogue # S-24732Used as a recording chamber in Ca imaging

Tags

Behavioral RecordingLaser AblationParamecia PreyRecording ChamberLighting SystemTime Lapse RecordingNeuron AssessmentCalcium Imaging