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Retro-orbital injections in mice


Intravenous vascular access is technically challenging in the adult mouse and even more challenging in neonatal mice. The authors describe the technique of retro-orbital injection of the venous sinus in the adult and neonatal mouse. This technique is a useful alternative to tail vein injection for the administration of non-tumorigenic compounds. The authors report that they have routinely used this technique in the adult mouse to administer volumes up to 150 μl without incident. Administration of retro-orbital injections is more challenging in neonatal mice but can reliably deliver volumes up to 10 μl.

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Figure 1: An anesthetized adult mouse placed on a protected warming device.
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Figure 5: Correct placement of the needle relative to the retro-orbital sinus, the eyeball and the back of the orbit.
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Figure 8: The blood vessels contributing to the retro-orbital sinus of the mouse.
Figure 9: A volume-rendered three-dimensional computed tomography scan of the head of a mouse (facing right) that received a retro-orbital injection of an intravascular contrast agent.
Figure 10: Section through the right eye and surrounding tissue of a 1-d-old mouse pup 30 min after administration of a retro-orbital injection (12.5× magnification; hematoxylin and eosin staining).
Figure 11: Section through the right eye and surrounding tissue of an adult mouse 30 min after administration of a retro-orbital injection (12.5× magnification; hematoxylin and eosin staining).


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We thank Brenda Klaunberg and Danielle Donahue from the NIH Mouse Imaging Facility and Darryl Leja from the NHGRI Intramural Publication Support Office for their assistance with this project. This work was carried out in partial fulfillment of the requirements for a PhD degree for T.Y. (Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel). This research was supported in part by the Intramural Research Program of the NHGRI, the National Institutes of Health.

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Correspondence to Shelley Hoogstraten-Miller.

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Yardeni, T., Eckhaus, M., Morris, H. et al. Retro-orbital injections in mice. Lab Anim 40, 155–160 (2011).

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