Ex Vivo Electroporation of Chick Embryo Cerebellar Slices: A Method to Introduce Plasmid DNA Encoding Green Fluorescent Protein to Visualize Granular Cell Development

Abstract

Source: Hanzel, M. et al. Ex Vivo Culture of Chick Cerebellar Slices and Spatially Targeted Electroporation of Granule Cell Precursors. J. Vis. Exp. (2015)

In this video, we perform electroporation of chick embryo cerebellum slices with reporter gene plasmids. The technique facilitates visualization of the development and migration of granule cell precursors to form granular layer.

Protocol

1. Electroporation of Slices

1. Construct an electroporation chamber by fixing the anode of an electroporator to the base of a 60 mm Petri dish with insulation tape. Add approximately 1 mL of HBSS to cover the electrode.
2. Place a 0.4 µm culture insert on top of the electrode covered in HBSS. Allow the culture insert to rest on the electrode making sure there is always contact between the insert and the electrode.
   NOTE: In this setup, the culture insert with the slices will rest upon the surface of the solution, maintaining the circuit but allowing spatial targeting of the cathode.
3. Transfer ident....

Results

Figure 1. The electroporation chamber set up. (A) A picture of the custom-made electroporation chamber. The chamber consists of an anode of an electroporator placed securely on the base of a 60 mm Petri dish. The dish cont.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Basal Medium Eagle (Gibco)	 Life Technologies	 41010-026
L-glutamine	 Sigma	 G7513
Penicillin/Streptomycin	 Sigma	 P4333
0.4 μm culture insert	 Millipore	 PICM0RG50
TSS20 Ovodyne electroporator	 Intracel	 01-916-02	Use 3 x 10 V 10 msec pulses for electroporation.