Tissue Collection for Systematic Phenotyping in the Mouse

Cristina Antal1, Marius Teletin1, Olivia Wendling1, Mounzer Dgheem1, Johan Auwerx1, Manuel Mark1

1 Université Louis Pasteur, Illkirch, France
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 29A.4
DOI:  10.1002/0471142727.mb29a04s80
Online Posting Date:  October, 2007
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Abstract

In this unit, a procedure for post‐mortem examination of mice and tissue collection is provided. This procedure is performed for post‐mortem analysis of anatomical defects (necropsy) and histological analysis and/or tissue collection destined for molecular biology applications. In both cases, tissue preservation is the major issue, but the way to achieve it depends on the objective. When histological analysis is the aim, tissue preservation is achieved by rapid transfer into fixative solutions. In contrast, molecular biology applications require rapid freezing of tissue samples to preserve mRNA integrity. Consequently, performing both procedures simultaneously may be at the expense of the final product quality. Curr. Protoc. Mol. Biol. 80:29A.4.1‐29A.4.23. © 2007 by John Wiley & Sons, Inc.

Keywords: phenotyping; mouse; tissue collection; histology; necropsy

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Necropsy for Histological Examination
  • Basic Protocol 2: Tissue Collection for Molecular Biology Applications
  • Commentary
  • Appendix: Trimming Organs and Defining Planes of Section
  • Key References
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Necropsy for Histological Examination

  Materials
  • Mice
  • CO 2 source
  • 70% ethanol
  • 10% (v/v) neutral buffered formalin (equivalent to 4% [w/v] formaldehyde; e.g., Carlo Erba)
  • Bouin's fixative solution (e.g., Carlo Erba)
  • 1× phosphate‐buffered saline (PBS; e.g., Sigma‐Aldrich)
  • Box with transparent walls and/or lids
  • Ruler
  • Electric shaver
  • Dissecting microscope equipped with a digital camera
  • Large containers with lids to hold fixing fluids (e.g., Labonord)
  • Dissection instruments, e.g., forceps, scissors (one pair dedicated to cutting bones), scalpels, razor blades
  • 1.5‐ml microcentrifuge tubes
  • Cork boards and needles
  • Plastic embedding cassettes (e.g., Labonord)
  • 5‐ml syringe and 25‐G needles
  • Balance (e.g., Fisher Bioblock Scientific)
  • 50‐ml plastic tubes with caps (e.g., Eppendorf)
  • Biopsy capsules
  • Petri dishes
  • Mouse coronal brain matrices (e.g., stainless‐steel for long‐term use or acrylic for sporadic use; Harvard Apparatus)

Basic Protocol 2: Tissue Collection for Molecular Biology Applications

  Materials
  • Mice
  • Liquid nitrogen
  • 70% ethanol
  • Dissection instruments (e.g., forceps, scissors)
  • 2‐ml sterile vials (e.g., Eppendorf)
  • Large styrofoam box with lid to hold the liquid nitrogen
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Figures

Videos

Literature Cited

   Brayton, C., Justice, M., and Montgomery, C.A. 2001. Evaluating mutant mice: Anatomic pathology. Vet. Pathol. 38:1‐19.
   Hebel, R. and Stromberg, M.W. 1986. Anatomy and Embryology of the Laboratory Rat. BioMed Verlag, Worthsee, Germany.
   Popesko, P., Raijtova, V., and Horak, J. 1992. A Colour Atlas of Anatomy of Small Laboratory Animals. Wolfe Publishing, London.
   Smith, R.S., John, S.W.M, Nishina, P.M., and Sundberg, J.P. 2002. Systematic Evaluation of the Mouse Eye: Anatomy, Pathology, and Biomethods. CRC Press, Boca Raton, Florida.
Internet Resources
  http://www.eumorphia.org/
  The EUMORPHIA Web site, which provides information about understanding human disease through mouse genetics.
  http://main.uab.edu/sites/ComparativePathology/links/
  The University of Alabama at Birmingham: Comparative Pathology Laboratory Web site.
  http://eulep.anat.cam.ac.uk/
  Pathbase Web site.
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