Neural Plate Cell Manipulation Through In Utero Nanoinjection in an Embryonic Mouse Model

Secure an anesthetized pregnant mouse with its exposed uterus containing pre-neurulation embryos. At this stage, the neural plate is open and directly exposed to amniotic fluid.

Position the selected embryos on a petri dish with an elastic membrane to stabilize them. Secure the embryos and apply saline for hydration.

Align the ultrasound probe over the embryo and measure the amniotic cavity diameter to calculate the optimal injection volume.

Position a nano-injection needle containing a high-titer lentiviral solution above the target embryo at a near-perpendicular angle.

Inject a precise small volume into the amniotic cavity, gently retract the needle and remove the ultrasound probe.

Gently push the embryos back inside the mouse.

The injected lentivirus fuses with neural plate cells, releasing its genetic material, which undergoes reverse transcription into DNA and is integrated into the host genome.

This enables stable gene manipulation in the neural plate cells, supporting potential therapeutic applications.

Using forceps, hold the tissue between the embryos and carefully pull out both uterine horns from the abdomen. Count and number the embryos, either from the ovary to the cervix or fr