Intrathoracic Injection: Compound Delivery to the Adult Zebrafish Heart

Abstract

Source: Bise, T., et al. Intrathoracic Injection for the Study of Adult Zebrafish Heart. J. Vis. Exp. (2019).

This video describes the technique of delivering compounds into the heart of adult zebrafish by using intrathoracic injection.

Protocol

1. Preparation of the Fish for Intrathoracic Injection Catch an adult zebrafish (Danio rerio) with a net and transfer it into the anesthetic solution. After 1−2 min, when fish stops swimming and the movement of operculum is reduced, touch the fish with a plastic spoon to make sure it does not react to any contact. Quickly and carefully transfer the fish with the spoon into the groove of the wet sponge, with ventral side up. The head of the fish should point away from the ope…

Representative Results

Figure 1: Intrathoracic (IT) injection in adult zebrafish. (A) Photograph of a pulled microinjection capillary with filament (6”, 1.0 mm in diameter) and values of the needle puller program used. (B) Photograph of a pulled microinjection capillary with filament (6”, 1.0 mm in diameter) filled up with 2.5 µL of solution containing 10% phenol red. The pulled tip of the needle is maximally 7 mm long. (C…

Materials

Macroscope (binocular)	M400		with Apozoom
Micro-injector femtojet	Eppendorf	5247 0034 77
Microloaders femtotips	Eppendorf	5242 956.003
Micropipette glass needles type C	WPI	TW100F-6	thin-wall capillary
Micropipette puller model P-87	Flaming/Brown	20081016	filament box 2.5 mm x 4.5 mm
Sponge	any	any	dim. carved sponge 7cm x 3 cm x 1 cm
Tricaine (Anestethic)	Sigma	E10521