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. Author manuscript; available in PMC: 2023 Sep 4.
Published in final edited form as: J Vis Exp. 2021 Jun 9;(172):10.3791/62568. doi: 10.3791/62568

Figure 2: In vivo cerebellar electroporation of granule neuron progenitors in P7 wildtype mouse pups.

Figure 2:

(A) Pups are anesthetized with 4% isoflurane delivered at a rate of 0.8L/min to ensure anesthesia throughout the injection of the DNA solution. Isoflurane is delivered at a rate of 0.8 L/min. (B) After sterilizing the mouse 3 times with betadine and 70% ethanol, an incision is made that spans the distance of the ears, revealing the hindbrain. (C) A magnified image of a white demarcation on the cranium, a landmark for the injection site. DNA construct should be injected within 1 mm above the mark; dotted lines outline the demarcation, and black arrow denotes the injection site. The ridges of the cerebellar vermis may be visible and can be useful for finding the injection site. (D) Tweezer-type electrode orientation for efficient electroporation. Plus (+) end must be oriented downwards to pull negatively charged DNA into the cerebellar parenchyma prior to administration of electrical pulses. (E) Test injection of 1 μL of a 0.02% Fast Green dye shows injection is localized to the middle of the cerebellar vermis between lobules 5–7.