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.
1. Assessment of Behavioral Consequences Following Laser Ablation
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) Components of the lighting system. A grey-colored paper, a glass diffuser, white LED ring light, diffuser, a custom-made spacer (cardboard), and diffuser with a hole at the center. (C) The lighting system assembled from the parts shown in B. (D) Recording chamber for the prey capture assay. A chamber (diameter: 20 mm; depth: 2.5 mm) was placed on a glass slide. The original top seal of the chamber was peeled off. A glass-cover was put on the recording chamber. (E) Raw image from a frame of the recorded movie. (F) A frame divided (pixel-by-pixel) by an average image in a movie. Note that the non-uniform background in lighting can be compensated by this image processing. (G) Paramecia extracted from a frame using the "Particle Analysis" function in Fiji. (H) Example of image in Fiji with an applied threshold. Paramecia appear bright, whereas the eyes of the zebrafish larva appear dark, and the background is grey. Changes in eye positions (i.e., angles with respect to the body axis) can be calculated, if necessary. (I) Representative experimental data of paramecium consumption in an olfactory-bulb-ablated control larva (OB) and a pretectum-ablated larva (PT). Pretectum ablation significantly reduced prey hunting ability while olfactory bulb ablation did not affect it.
Imageing Chambers | Grace Bio-Labs | CoverWell Imaging Chambers PCIA-2.5 | Used as a behavioral recording chamber |
ORCA-Flash4.0 | Hamamatsu Photonics | Model:C11440-22CU | A scientific CMOS camera |
SZX7 | Olympus | Stereoscope | |
A ring LED light | CCS | Model: LDR2-100SW2-LA | White LED |
Secure-Seal Hybridization Chamber Gasket, 8 chambers, 9 mm diameter x 0.8 mm depth | Molecular Probes | Catalogue # S-24732 | Used as a recording chamber in Ca imaging |