Standardized Post‐Mortem Examination and Fixation Procedures for Mutant and Treated Mice

Cristina Antal1, Stéphanie Muller2, Olivia Wendling3, Yann Hérault3, Manuel Mark4

1 Institut d'Histologie, Faculté de Médecine, Strasbourg, France, 2 Mouse Clinical Institute, Illkirch, France, 3 Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), CNRS/INSERM/Université Louis Pasteur, Illkirch, France, 4 Hôpital Universitaire de Strasbourg, Strasbourg, France
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo100118
Online Posting Date:  March, 2011
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A procedure for post‐mortem examination (or necropsy) of mice is provided. The aim is to obtain a “holistic” picture of organs and systems at the anatomical and histological levels. The major issue is tissue preservation, which is achieved by rapid transfer into a fixative solution, usually neutral buffered formalin. Fixation is the first of the four basic steps in histopathological analyses of tissues, which also include embedding, sectioning, and staining. The protocols provided here describe routine methods for tissue fixation, as these methods are integral parts of any necropsy procedure. There is also a Strategic Planning section that addresses the overall approach to histopathological evaluation, as well as specifics such as age and gender of the mice, cohort size, and controls. Curr. Protoc. Mouse Biol. 1:17‐53. © 2011 by John Wiley & Sons, Inc.

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

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Table of Contents

  • Introduction
  • Strategic Planning
  • Basic Protocol 1: Necropsy for Histopathological Analysis
  • Basic Protocol 2: Fixation for Routine Histopathological Analyses with Formalin
  • Alternate Protocol 1: Fixation for Routine Histopathological Analyses with Bouin's Solution
  • Support Protocol 1: Trimming Organs and Defining Planes of Section
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
  • Tables
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Basic Protocol 1: Necropsy for Histopathological Analysis

  • Mice (see Strategic Planning)
  • 10% (v/v) neutral buffered formalin (equivalent to 4% [w/v] formaldehyde; see recipe)
  • 70% ethanol (see recipe)
  • Plastic embedding cassettes (e.g., Fisher Scientific)
  • Large (≥1 liter) containers with lids (e.g., Fisher Scientific)

Basic Protocol 2: Fixation for Routine Histopathological Analyses with Formalin

  • Mice (see Strategic Planning)
  • Bouin's fixative solution (e.g., VWR International LLC )
  • 70% ethanol (see recipe)
  • Plastic embedding cassettes (e.g., Fisher Scientific)
  • Large containers with lids (e.g., Fisher Scientific)
  • Rocking platform
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Literature Cited

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Internet Resources
  The EUMORPHIA Web site, provides information about understanding human disease through mouse genetics.
  Pathbase, a database of histopathology photomicrographs and macroscopic images derived from mutant or genetically manipulated mice.
  Provides an electronic atlas for the anatomy and histology of the mouse.
  Provides an electronic atlas for the histology of the mouse.
  A workbook with a set of microscope slides that illustrate the basic tissues and organs of the body.
  Histosearch is a search engine that searches over 20,000 Web pages from histology‐related sites.
  StainsFile is a useful site with general information and resources for histology.
  Information on formaldehyde fixatives at the Southwest Environmental Health Sciences Center (SWEHSC), University of Arizona College of Pharmacy.
  The Canadian Council on Animal Care (CCAC) is the national organization responsible for setting and maintaining standards for the care and use of animals in science in Canada.
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