Feb 16, 2025

Public workspaceRetro-orbital injection of AAV in mice

 Forked from Retro-orbital injection of AAV in mice
DOI
dx.doi.org/10.17504/protocols.io.bp2l6drpdvqe/v1
  • 1Caltech
Chelsie Steele
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DOI: dx.doi.org/10.17504/protocols.io.bp2l6drpdvqe/v1
Protocol Citation: Chelsie Steele, Yujie Fan 2025. Retro-orbital injection of AAV in mice. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6drpdvqe/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 07, 2025
Last Modified: February 16, 2025
Protocol Integer ID: 117848
Keywords: Mice, Retro-orbital injections, ASAPCRN
Funders Acknowledgements:
Aligning Science Across Parkinson’s
Grant ID: ASAP-020495
Abstract
This protocol is following:

CITATION
Tal Yardeni, Michael Eckhaus, H. Douglas Morris, Marjan Huizing, Shelley Hoogstraten-Miller (2011). Retro-orbital injections in mice. LabAnimal.
LINK
10.1038/laban0511-155





Guidelines
Please adhere to your institue IACUC guidelines of animal care.
Materials
MATERIALS
Reagent29G x 12.7mm 0.3ml syringeUlticareCatalog #09230
STEP MATERIALS
Reagent29G x 12.7mm 0.3ml syringeUlticareCatalog #09230
It is important that the needel used has low friction so that the injection is at a slow and consistent rate.
Injection
Injection
Prepare virus dilution with sterile DPBS. Injection is 100 μL per mouse.



Take one mouse from the cage into an induction chamber and start isoflurane at 3-5% with flow rate of 0.8-1L/min
After breathing rate of the mouse has slowed (~1/s), indicating sufficient anesthetic depth, you can begin working with the mouse
Expose the eye ball with two fingers (above and below) and make sure the you are not pressing on the trachea and the mouse's breathing is regular.
Critical
Use a 31 G insulin syringe at a 30-degree angle beveled downwards until you hit the bone, retract a bit and slowly inject. See cited paper for more details.


Equipment
U-100 BD Ultra-Fine™ Short Insulin Syringes
NAME
Syringe and neecdle
TYPE
BD Medical
BRAND
BD328438
SKU
https://us.vwr.com/store/catalog/product.jsp?catalog_number=BD328438
LINK

Critical
Remove the needle slowly and press the eyelid shut for a few seconds.
Prior to recovery, apply 1-2 drops 0.5% ophthalmic Proparacaine to the injected eye

Optional