Mouse Necropsy

Piper M. Treuting1, Jessica M. Snyder1

1 Department of Comparative Medicine & Comparative Pathology Program, School of Medicine, University of Washington, Seattle, Washington
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo140296
Online Posting Date:  September, 2015
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Abstract

Necropsy (also known as autopsy) is the post‐mortem dissection of bodies after euthanasia or death and is a scientific examination conducted to observe and dissect the organs, collect tissues, and determine the extent of grossly evident disease. Research necropsies are conducted to obtain specific samples tailored according to study objectives. Diagnostic necropsy may be undertaken when unexpected illness or death occurs. The systematic collection of samples at necropsy is the critical first step in generating morphologic data from animal models. The morphologic (anatomic and histologic) data generates information on changes in cells, tissues, organs, and organ systems providing context for phenotypes (functional and morphological) to the level of the whole organism. Optimal insight into phenotype or pathophysiologic mechanisms is obtained when morphologic data is coupled with laboratory, medical, and molecular findings. This protocol provides a standard for an efficient routine mouse necropsy with brief comments on advanced or alternative techniques. © 2015 by John Wiley & Sons, Inc.

Keywords: rodent; autopsy; histopathology; fixation; disease models; phenotyping

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Basic Necropsy of the Mouse
  • Alternate Protocol 1: Abbreviated Necropsy for Mice
  • Commentary
  • Figures
     
 
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Materials

Basic Protocol 1: Basic Necropsy of the Mouse

  Materials
  • Mouse
  • 70% to 95% ethanol in a dip‐container or spray bottle
  • 10% (v/v) neutral buffered formalin (fixative; e.g., VWR, cat. no. 16004‐124)
  • Specimen containers
  • Embedding cassettes
  • #2 pencil
  • Necropsy record
  • Optional: supplies for blood collection
  • Dissection board
  • Dissection pins
  • Forceps
  • Surgical scissors
    • Sharp for most dissection
    • Less sharp or older for tough tissue
  • Digital scale (readability 0.01 g)
  • Ruler (metric)
  • Optional: supplies for preparing intestinal Swiss roll
  • 3‐ml syringe
  • 18‐ or 21‐G needle
  • Dissecting microscope or magnifying lens
  • Optional: biopsy bag
  • Optional: digital camera for documentation

Alternate Protocol 1: Abbreviated Necropsy for Mice

  Materials
  • Specimen container filled with 10% (v/v) neutral buffered formalin or 450‐ml prefilled formalin container (e.g., VWR, cat. no. 16004‐124)
  • Necropsy record
  • Surgical scissors
  • Forceps
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Figures

Videos

Literature Cited

Literature Cited
  Bevilacqua, G., Bosman, F., Dassesse, T., Hofler, H., Janin, A., Langer, R., Larsimont, D., Morente, M.M., Riegman, P., Schirmacher, P., Stanta, G., Zatloukal, K., Caboux, E., and Hainaut, P. 2010. The role of the pathologist in tissue banking: European consensus expert group report. Virchows Arch. 456:449‐454. doi: 10.1007/s00428‐010‐0887‐7.
  Bolon, B., Couto, S., Fiette, L., and Perle, K.L. 2012. Internet and print resources to facilitate pathology analysis when phenotyping genetically engineered rodents. Vet. Pathol. 49:224‐235. doi: 10.1177/0300985811415709.
  Bolon, B., Garman, R.H., Pardo, I.D., Jensen, K., Sills, R.C., Roulois, A., Radovsky, A., Bradley, A., Andrews‐Jones, L., Butt, M., and Gumprecht, L. 2013. STP position paper: Recommended practices for sampling and processing the nervous system (brain, spinal cord, nerve, and eye) during nonclinical general toxicity studies. Toxicol. Pathol. 41:1028‐1048. doi: 10.1177/0192623312474865.
  Brayton, C., Justice, M., and Montgomery, C.A. 2001. Evaluating mutant mice: Anatomic pathology. Vet. Pathol. 38:1‐19. doi: 10.1354/vp.38‐1‐1.
  Cardiff, R.D., Miller, C.H., and Munn, R.J. 2014a. Limited mouse necropsy. Cold Spring Harb. Protoc. doi: 10.1101/pdb.prot073395.
  Cardiff, R.D., Miller, C.H., and Munn, R.J. 2014b. Mouse tissue fixation. Cold Spring Harb. Protoc. doi: 10.1101/pdb.prot073403.
  Cardiff, R.D., Rosner, A., Hogarth, M.A., Galvez, J.J., Borowsky, A.D., and Gregg, J.P. 2004. Validation: The new challenge for pathology. Toxicol. Pathol. 32:31‐39. doi: 10.1080/01926230490424662.
  de Assis, S., Warri, A., Cruz, M.I., and Hilakivi‐Clarke, L. 2010. Changes in mammary gland morphology and breast cancer risk in rats. J. Vis. Exp. pii:2260. doi: 10.3791/2260.
  Donovan, J. and Brown, P. 2006. Blood collection. Curr Protoc. Immunol. 73:1.7.1‐1.7.9. doi: 10.1002/0471142735.im0107s73.
  Kittel, B., Ruehl‐Fehlert, C., Morawietz, G., Klapwijk, J., Elwell, M.R., Lenz, B., O'Sullivan, M.G., Roth, D.R., and Wadsworth, P.F. 2004. Revised guides for organ sampling and trimming in rats and mice–Part 2. A joint publication of the RITA and NACAD groups. Exp. Toxicol. Pathol. 55:413‐431. doi: 10.1078/0940‐2993‐00349.
  Knoblaugh, S.E., Randolph‐Habecker, J., and Rath, S. 2012. Necropsy and Histology. In Comparative Anatomy and Histology: A Mouse and Human Atlas (P.M. Treuting and S.M. Dintzis, eds.) pp. 15‐37. Academic Press, London.
  McInnes, E.F. 2011. Background Lesions in Laboratory Animals: A Color Atlas. Elsevier Science Publishing, New York.
  Moolenbeek, C. and Ruitenberg, E.J. 1981. The “Swiss roll”: A simple technique for histological studies of the rodent intestine. Lab. Anim. 15:57‐59. doi: 10.1258/002367781780958577.
  Morawietz, G., Ruehl‐Fehlert, C., Kittel, B., Bube, A., Keane, K., Halm, S., Heuser, A., and Hellmann, J. 2004. Revised guides for organ sampling and trimming in rats and mice—Part 3. A joint publication of the RITA and NACAD groups. Exp. Toxicol. Pathol. 55:433‐449. doi: 10.1078/0940‐2993‐00350.
  Parkinson, C.M., O'Brien, A., Albers, T.M., Simon, M.A., Clifford, C.B., and Pritchett‐Corning, K.R. 2011. Diagnostic necropsy and selected tissue and sample collection in rats and mice. J. Vis. Exp. pii:2966. doi: 10.3791/2966.
  Ruehl‐Fehlert, C., Kittel, B., Morawietz, G., Deslex, P., Keenan, C., Mahrt, C.R., Nolte, T., Robinson, M., Stuart, B.P., and Deschl, U. 2003. Revised guides for organ sampling and trimming in rats and mice–Part 1. Exp. Toxicol. Pathol. 55:91‐106. doi: 10.1078/0940‐2993‐00311.
  Scudamore, C.L.C. 2013. A Practical Guide to the Histology of the Mouse. John Wiley & Sons, New York.
  Scudamore, C.L., Busk, N., and Vowell, K. 2014. A simplified necropsy technique for mice: Making the most of unscheduled deaths. Lab. Anim. 48:342‐344. doi: 10.1177/0023677214536555.
  Son, A.I., Sokolowski, K., and Zhou, R. 2013. Cryosectioning. Methods Mol. Biol. 1018:301‐311.
  Treuting, P.M. and Dintzis, S.M. (eds.) 2012. Comparative Anatomy and Histology: A Mouse and Human Atlas. Academic Press, London.
Key References
  Bolon et al., 2012. See above.
Internet Resources
  http://tvmouse.ucdavis.edu/virtualNecropsy/
  Images showing necropsy of the mouse.
  http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3211129/
  Video and protocol for necropsy of the mouse and rat.
  http://research.fhcrc.org/fero/en/fero‐lab‐protocols/complete‐mouse‐necropsy.html
  Protocol from Matthew Fero laboratory detailing euthanasia, fixatives, and necropsy of the mouse.
  http://ctrgenpath.net/static/atlas/mousehistology/
  Images from the Atlas of Laboratory Mouse Histology.
  http://courses.jax.org/
  Course and conference listings offered by The Jackson Laboratory, including the Annual Workshop on Pathology of Mouse Models of Human Diseases.
  http://www.hopkinsmedicine.org/mcp/PHENOCORE/courseCURRENT.html
  Mouse Pathobiology and Phenotyping Short Course offered by Johns Hopkins Medicine.
  http://www.cldavis.org/
  Meetings and syllabi offered by the Charles Louis Davis, D.V.M. Foundation, including the Pathology of Laboratory Animals course.
  http://www.informatics.jax.org/external/festing/mouse/STRAINS.shtml
  List of major mouse strains with information and links.
  http://www.informatics.jax.org/mgihome/nomen/index.shtml
  Mouse nomenclature: rules for naming mouse and rat strains, genes, alleles, and mutations.
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