Zebrafish Brain Dissection: A Technique of Fish Neurobiology

Abstract

Source: Lopez-Ramirez M. A. et. al., Isolation and Culture of Adult Zebrafish Brain-derived Neurospheres. J. Vis. Exp. (2016).

This video describes the technique of microdissection of the brain from an adult zebrafish. This method helps in obtaining whole brain derived neuorospheres that can be studied further.

Protocol

1. Dissection of the Adult Zebrafish Brain Prepare a dissection bed by filling a 100 mm x 15 mm Petri dish with gel ice packs. Then place the lid on the petri dish and incubate at -20 °C until the gel freezes. On top of the lid place a square of clean filter paper and wrap both the filter paper and petri dish with plastic film. Clean and sterilize all the microdissection instruments by 70% ethanol or heat before each use. Place all sterilized dissection instruments near the dissecting microscop…

Representative Results

Figure 1: Differentiation of Zebrafish Brain-derived Neurospheres. (A-C, E) Phase contrast images of whole brain-derived neurospheres cultured in the Z-differentiation medium during 1 (A, DiVd1), 3 (B, DiVd3) and 4 (C and D, DiVd4) days. (E) Dorsal view of the who…

Materials

DMEM/F12 1x	Life Technologies	11330-032
Penicillin-streptomycin	Life Technologies	15140-122
Tricaine MS-222	Sigma	A5040	stock solution of 4 mg/ml