Home › Neuroscience › Ultrasound-Guided Microinjection into the Mouse Forebrain In Utero at E9.5
Neuroscience JoVE (Open Access) Citable · DOI

Ultrasound-Guided Microinjection into the Mouse Forebrain In Utero at E9.5

DOI: 10.3791/2047-v
What you'll learn
  • ✓ Perform ultrasound-guided microinjection into embryonic mouse forebrain at E9.5
  • ✓ Expose and visualize embryos in utero using high-frequency ultrasound imaging
  • ✓ Load retroviral vectors into microinjection needles with precision
  • ✓ Deliver viral vectors to target brain regions during early development
Protocol

Biopharma Insights In utero survival surgery in mice permits the molecular manipulation of gene expression during development. Here we describe the use of high-frequency ultrasound imaging to guide the injection of retroviral vectors into the mouse brain at embryonic day (E) 9.5.

Difficulty
advanced
Total time
~2–3 hours per pregnant dam (includes anesthesia, surgery, imaging, injections, and closure)
Model organism
Mouse (pregnant dam, embryonic day 9.5)
Biosafety
BSL-2

Steps

1
Expose embryos through uterine surgical incision

Perform survival surgery on anesthetized pregnant mouse to carefully expose the uterine horns and position embryos for visualization and injection.

▶ 01:56
2
Load retroviral vector into microinjection needle

Prepare the microinjection needle by loading retroviral vector solution into the needle tip using standard microinjection pipetting techniques.

▶ 03:45
3
Inject virus into forebrain under ultrasound guidance

Use real-time high-frequency ultrasound imaging to visualize the embryonic brain and guide precise microinjection of retroviral vector into the target forebrain region.

▶ 04:49
4
Assess injection success and tissue viability

Evaluate representative results by examining successful vector delivery, embryo positioning, and overall tissue integrity post-injection.

▶ 06:25
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