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High Precision Stereotaxic Surgery in Mice
Publication Name:
Current Protocols in Neuroscience
Unit Number:
Appendix 4A
DOI:
10.1002/0471142301.nsa04as14
Online Posting Date:
May, 2001 Abstract
Table of Contents
- Unit Introduction
- Basic Protocol: Site-Specific Central Microinjection in the Anesthetized Mouse
- Alternate Protocol: Cannula Implantation and Central Microinjection in the Behaving Mouse
- Support Protocol: Cannula,Wire Plug, and Injection Needle Manufacture
- Reagents and Solutions
- Commentary
- Bibliography
- Figures
Materials
Basic Protocol: Site-Specific Central Microinjection in the Anesthetized Mouse
Materials
- Ketamine/xylazine cocktail (see
- 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: Cannula Implantation and Central Microinjection in the Behaving Mouse
Additional Materials (also see Basic Protocol )
- 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
- Cannula holder
- Injection needle (see
- Polyethylene tubing (0.58-mm i.d.)
- Micro-syringe
- Syringe pump
Support Protocol: 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.
Looking for materials? Suppliers Guide
Figures
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Figure A.4A.1Diagram of the cannula, wire plug, and injector needle. (A) Cannula: stainless steel hypodermic tubing is cut to length and abraded with an abrasive wheel. (B) Wire plug: a short stainless steel cap is soldered onto a length of smaller diameter tubing or solid-core wire such that when it is seated within the cannula, (C) its tip protrudes. (D) Injection needle: a length of stainless steel tubing is soldered onto smaller diameter tubing. Like the wire plug, the tip of the injection needle extends beyond the end of the cannula. However, the smaller diameter tubing is left open so that it can be attached to a length of polyethylene tubing and fluids can be administered through it by means of a syringe pump.
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 | |
| In the authors' experience, the most accurate mouse stereotaxic atlas. | |
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