High Precision Stereotaxic Surgery in Mice

Jaime Athos1, Daniel R. Storm1

1 University of Washington, Seattle, Washington
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Appendix 4A
DOI:  10.1002/0471142301.nsa04as14
Online Posting Date:  May, 2001
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Abstract

This unit provides protocols for cannulation and site‐specific central microinjection of mice using a recently developed high‐precision stereotaxic frame. The construction of cannulae, wire plugs and injection needles are also described.

     
 
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Table of Contents

  • Basic Protocol 1: Site‐Specific Central Microinjection in the Anesthetized Mouse
  • Alternate Protocol 1: Cannula Implantation and Central Microinjection in the Behaving Mouse
  • Support Protocol 1: Cannula,Wire Plug, and Injection Needle Manufacture
  • Reagents and Solutions
  • Commentary
  • Figures
     
 
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Materials

Basic Protocol 1: Site‐Specific Central Microinjection in the Anesthetized Mouse

  Materials
  • Ketamine/xylazine cocktail (see recipe)
  • Hair shaver or depilatory cream (optional)
  • Ophthalmic ointment or mineral oil
  • Betadine
  • Bone wax
  • Mouse stereotaxic frame (Cartesian Research) equipped with:
  •  Sighting scope
  •  Drill with #74 bit
  •  Monoinjector with 24‐G Hamilton syringe (model #8800 for injections of up to 5 µl)
  • Heating pad
  • Forceps
  • Scalpel
  • Surgical scissors
  • Agricola‐style retractors (Fine Science Tools)
  • Cotton swabs
  • Sutures

Alternate Protocol 1: Cannula Implantation and Central Microinjection in the Behaving Mouse

  • Filter‐sterilized food coloring (dark colors such as blue, black, or green work best)
  • Cyanoacrylate glue and accelerator (hobby supplies store)
  • Dental acrylic
  • Injectate
  • Cannula and wire plug (see protocol 3)
  • Cannula holder
  • Injection needle (see protocol 3)
  • Polyethylene tubing (0.58‐mm i.d.)
  • Micro‐syringe
  • Syringe pump

Support Protocol 1: Cannula,Wire Plug, and Injection Needle Manufacture

  Materials
  • Ethanol
  • Acid flux (welding supply store)
  • Dremel tool with abrasive wheel and cutting disk (Small Parts)
  • Small bench vise
  • 24‐G thin‐walled stainless steel hypodermic tubing (Small Parts)
  • 30‐G stainless steel hypodermic tubing and/or solid core wire (Small Parts)
  • Hemostats
  • Soldering iron (40‐W minimum)
  • Silver bearing solder (welding supply store)
  • Outside micrometer (Small Parts)
NOTE: To further reduce the damage to the brain, smaller gauge cannulae, wire plugs and injectors can be used. For example, 28‐G thin‐walled tubing and 33‐G tubing can be substituted for the 24‐G thin‐walled tubing and 30‐G tubing recommended here. The smaller tubing is more prone to bending and occlusion, however, and will increase the difficulty of parts manufacture and animal infusion.
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Figures

Videos

Literature Cited

Literature Cited
   Slotnick, B.M. and Leonard, C.M. 1975. A Stereotaxic Atlas of the Albino Mouse Forebrain. DHEW Publication (ADM) 75‐100, U.S. Government Printing Office, Rockville, Md.
Key References
   Flecknell, P.A. 1996. Laboratory Animal Anaesthesia: A Practical Introduction for Research Workers and Technicians. Academic Press, San Diego.
  A good discussion of alternative methods of anesthesia including Avertin, pentobarbital, and halothane.
   Slotnick and Leonard, 1975. See above.
  In the authors' experience, the most accurate mouse stereotaxic atlas.
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