Care and Handling of Laboratory Mice

John Donovan1, Patricia Brown2

1 Bioresources Consulting, Weston, Vermont, 2 National Institutes of Health, Bethesda, Maryland
Publication Name:  Current Protocols in Microbiology
Unit Number:  Appendix A.3N
DOI:  10.1002/9780471729259.mca03ns31
Online Posting Date:  November, 2013
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Abstract

The importance of laboratory mice in experimental microbiology and biomedical research is indisputable, and their care and handling is a contributing factor to the quality of the science resulting from their use. This unit provides guidelines for practical housing, handling, and the methodology for sample collection to ensure animal health and minimize variable experimental parameters. Curr. Protoc. Microbiol. 31:A.3N.1‐A.3N.18. © 2013 by John Wiley & Sons, Inc.

Keywords: mouse; mouse housing; mouse handling; mouse blood collection; parenteral injections

     
 
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Table of Contents

  • Introduction
  • Caging
  • Feed
  • Bedding
  • Water
  • Sanitation
  • Provisions for Off‐Hour Care
  • Environmental Monitoring and Maintenance
  • Social Housing and Environmental Enrichment
  • Protocols: Handling and Restraint
  • Basic Protocol 1: Mouse Handling and Manual Restraint
  • Alternate Protocol 1: Rodent Restrainers
  • Protocols: Anesthesia
  • Basic Protocol 2: Injectable Anesthesia for Mouse
  • Protocols: Parenteral Injections
  • Basic Protocol 3: Intramuscular Injection of Mouse
  • Basic Protocol 4: Intradermal Injection of Mouse
  • Basic Protocol 5: Subcutaneous Injection of Mouse
  • Basic Protocol 6: Intravenous Injection of the Mouse
  • Basic Protocol 7: Intraperitoneal Injection of Mouse
  • Protocols: Blood Collection
  • Basic Protocol 8: Blood Collection from Orbital Sinus or Plexus of Mouse
  • Basic Protocol 9: Blood Collection from Tail Vein of Mouse Using Microhematocrit Tube
  • Alternate Protocol 2: Blood Collection from Tail Vein of Mouse Using Centrifuge Tube
  • Basic Protocol 10: Blood Collection from Axillary Plexus of Mouse
  • Basic Protocol 11: Cardiac Puncture of Mouse
  • Protocols: Euthanasia
  • Basic Protocol 12: Carbon Dioxide Asphyxiation
  • Basic Protocol 13: Pentobarbital Overdose
  • Alternate Protocol 3: Cervical Dislocation of Mouse
  • Disclaimer
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Mouse Handling and Manual Restraint

  Materials
  • Anesthetic of choice (Table 3.0.3)
  • Beaker
  • Laboratory balance for weighing animals
  • 1‐ or 3‐ml syringe with 22‐G needle
  • Additional reagents and equipment for handling and restraint ( protocol 1) and intraperitoneal injection ( protocol 9)

Alternate Protocol 1: Rodent Restrainers

  Materials
  • Injectate
  • 1‐ to 3‐ml syringe with 22‐ to 30‐G needle

Basic Protocol 2: Injectable Anesthesia for Mouse

  Materials
  • 70% ethanol
  • Injectate
  • Clippers with no. 40 blade
  • Gauze sponge or swab
  • 1‐ to 3‐ml syringe with 25‐ to 30‐G needle
  • Additional reagents and equipment for handling and restraint (see protocol 1 and protocol 2) and anesthesia (see Basic Protocol protocol 32 or protocol 43)

Basic Protocol 3: Intramuscular Injection of Mouse

  Materials
  • Injectate
  • 1‐ to 3‐ml syringe with 22‐ to 30‐G needle

Basic Protocol 4: Intradermal Injection of Mouse

  Materials
  • Injectate
  • 70% ethanol
  • 1‐ml syringe with 25‐ to 30‐G needle
  • Restrainer (see protocol 2)
  • Heat lamp or beaker containing warm water
  • Gauze sponge or swab

Basic Protocol 5: Subcutaneous Injection of Mouse

  Materials
  • Injectate
  • 1‐ or 3‐ml syringe with 22‐ to 25‐G needle

Basic Protocol 6: Intravenous Injection of the Mouse

  Materials
  • Sterile saline or phosphate‐buffered saline (PBS; appendix 2A)
  • Microhematocrit tube
  • Gauze sponge or swab
  • Additional reagents and equipment for handling and restraint (see protocol 1) and anesthesia (see Basic Protocol protocol 32 or protocol 43)

Basic Protocol 7: Intraperitoneal Injection of Mouse

  Materials
  • Heat lamp or beaker containing warm water
  • 25‐ to 30‐G needle
  • Microhematocrit tube
  • Gauze sponge
  • Additional reagents and equipment for handling and restraint (see protocol 1 or protocol 2)

Basic Protocol 8: Blood Collection from Orbital Sinus or Plexus of Mouse

  Additional Materials (also see protocol 10)
  • Scalpel blade (no. 10)
  • 12‐ to 50‐ml plastic centrifuge tube

Basic Protocol 9: Blood Collection from Tail Vein of Mouse Using Microhematocrit Tube

  Materials
  • Surgical board
  • Scalpel or razor blade
  • Thumb forceps
  • Surgical scissors
  • Additional reagents and equipment for anesthesia (see Basic Protocol protocol 32 or protocol 43)

Alternate Protocol 2: Blood Collection from Tail Vein of Mouse Using Centrifuge Tube

  Materials
  • 1‐ to 3‐ml syringe (mouse) with 20‐ to 22‐G needle
  • Additional reagents and equipment for anesthesia (see Basic Protocol protocol 32 or protocol 43)

Basic Protocol 10: Blood Collection from Axillary Plexus of Mouse

  Materials
  • 100% CO 2 (tank or house carbon dioxide) with pressure‐reducing regulator and liter flow meter
  • CO 2 chamber with lid

Basic Protocol 11: Cardiac Puncture of Mouse

  Materials
  • Sodium pentobarbital (60 mg/kg)
  • 1‐ to 3‐ml syringe with 20‐ to 25‐G needle
  • Additional reagents and equipment for handling and restraint (see protocol 1 or protocol 2) and intraperitoneal injection (see protocol 9)

Basic Protocol 12: Carbon Dioxide Asphyxiation

  Materials
  • Pencil or metal rod of similar size
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Figures

Videos

Literature Cited

  Committee for the Update of the Guide for the Care and Use of Laboratory Animals. 2011. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Institute for Laboratory Animal Research, Division on Earth and Life Studies, National Research Council, The National Academies Press, Washington, D.C. http://grants.nih.gov/grants/olaw/Guide‐for‐the‐Care‐and‐Use‐of‐Laboratory‐Animals.pdf.
  Fox, J.G., Anderson, L.C., Loew, F.M., and Quimby, F.W. (eds.). 2002. Laboratory Animal Medicine, 2nd ed., pp. 179‐180. Academic Press, San Diego.
  Gay, W.I. 1986. Methods of Animal Experimentation, Vol. VII, pp. 31‐65. Academic Press, San Diego.
  ILAR Committee for the Update of the Guide for the Care and Use of Laboratory Animals. 2011. Guide for the Care and Use of Laboratory Animals, 8th Edition. Institute for Laboratory Animal Research, Division on Earth and Life Studies, National Research Council, National Academies Press, Washington, D.C.
  Leary, S., Underwood, W., Anthony, R., Cartner, S., Corey, D., Grandin, T., Greenacre, C., Gwaltney‐Brant, S., McCrackin, M.A., Meyer, R., Miller, D., Shearer, S., and Yanong, R. 2013. AVMA Guidelines for the Euthanasia of Animals. https://www.avma.org/KB/Policies/Documents/euthanasia.pdf.
  Russell, W.M.S. and Burch, R.L. 1959. The Principles of Humane Experimental Technique. Methuen, London.
  Suckow, M.A., Danneman, P., and Brayton, C. 2001. Experimental methodology. In The Laboratory Mouse, p. 113. CRC Press, Boca Raton, Fla.
  Tuffery, A.A. 1987. Laboratory Animals: An Introduction for New Experimenters, pp. 225‐226. John Wiley & Sons, Chichester, U.K.
Key References
  Fox et al., 2002. See above.
  Provides additional details for injection methods.
  Gargiulo, S., Greco, A., Gramanzini, M., Esposito, S., Affuso, A., Brunetti, A., and Vesce, G. 2012. Mice anesthesia, analgesia and care, part I: Anesthetic considerations in preclinical research ILAR J. 53:E55–E69. http://ilarjournal.oxfordjournals.org/content/53/1/E55.full.pdf+html.
  White, W.J. and Field, K.J. 1987. Anesthesia and surgery of laboratory animals. In The Veterinary Clinics of North America, Small Animal Practice, Exotic Pet Medicine, 17:5 (J.E. Harkness, ed.) pp. 989‐1017. W.B. Saunders, Philadelphia.
  Wixson, S.K. and Smiler, K.L. 1997. Anesthesia and analgesia in rodents. In Anesthesia and Analgesia in Laboratory Animals (D.F. Kohn, S.K. Wixson, W.J. White, and G.J. Benson, eds.) pp. 165‐200. Academic Press, San Diego.
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