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本文内容

  • Overview
  • 研究方案
  • 结果
  • 材料
  • 参考文献

Overview

This video describes in vivo electroporation, which is a technique for DNA delivery into ependymoglial cells in adult zebrafish.

研究方案

1. Electroporation

  1. Remove the fish from the injection set-up while still holding it in the sponge.
  2. Immerse the inner side of the tip of the electrodes in the ultrasound gel.
  3. Cover the fish telencephalon with a small amount of ultrasound gel.
  4. Position the fish head between the electrodes, placing the positive electrode at the ventral side of the fish’s head and the negative electrode on the dorsal side (Figure 1C), while still holding the fish’s body in the sponge. This sets the direction of the flow of the current necessary to electroporate ependymoglia positioned at subventricular zone.
  5. Press the electrodes gently and precisely against the telencephalon (Figure 1 C). Administer the current with the foot pedal. Hold the electrodes in place until all five pulses are finished.

结果

figure-results-25
Figure 1: Schematic representation of coronal section of the everted zebrafish telencephalon. (A) Scheme of a coronal section of zebrafish telencephalon, highlighting the position of ependymoglial cells, which are lining ventricular surface and building the ventral ventricular wall. Dorsal ependymal layer is bridging the two hemispheres and covering the ventricle (V), located in between two cell...

材料

NameCompanyCatalog NumberComments
Ultrasound gelSignaGel, Parker laboratories INC15-60Electrode Gel
BTX Tweezertrodes ElectrodesPlatinum Tweezertrode, BTX Harvard Apparatus45-04861mm diameter
Electroporation deviceBTX ECM830 Square Wave Electroporation System, BTX Harvard Apparatus45-0662

参考文献

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