Histopathological Analysis of the Respiratory Tract

Cheryl L. Scudamore1, Elizabeth F. McInnes2

1 Mary Lyon Centre, MRC Harwell, 2 Cerberus Sciences, Thebarton, South Australia
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo140094
Online Posting Date:  December, 2014
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Abstract

The basic anatomy of the mouse respiratory system is similar to that of other mammals and can be usefully examined under the light microscope in phenotyping studies, inhalation toxicity studies, and studies involving mouse models of human disease. In many studies, however, only the lungs are examined, leaving the possibility that phenotypic information from the majority of the conducting airways is lost. This unit provides standard approaches for tissue collection at necropsy and subsequent selection of a range of respiratory tissues for histological and pathological analysis. The major anatomical features to be found in each section are highlighted, and potential artifacts and methods to avoid them are discussed. © 2014 by John Wiley & Sons, Inc.

Keywords: lung; nasal cavity; larynx; pathology; histology

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Inflation Fixation of Lungs
  • Alternate Protocol 1: Perfusion Fixation of Lungs
  • Support Protocol 1: Fixation for Standard Histological Staining or Immunohistochemistry
  • Basic Protocol 2: Fixation of Nasal Cavity
  • Basic Protocol 3: Sampling Respiratory Tract Tissues for Histology
  • Reagents and Solutions
  • Commentary
  • Figures
     
 
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Materials

Basic Protocol 1: Inflation Fixation of Lungs

  Materials
  • Mouse
  • 70% (v/v) alcohol
  • 10% neutral buffered formalin (NBF; see recipe)
  • Dissection instruments, depending on user preference
  • Down‐draft workstation or flow cabinet
  • 5‐ml syringe with 26‐G needle

Alternate Protocol 1: Perfusion Fixation of Lungs

  Materials
  • Mouse
  • Phosphate‐buffered saline (PBS; e.g., Oxoid, cat. no. BR0014G)
  • Fixative: 10% NBF (see recipe) or 4% (w/v) paraformaldehyde (see recipe)
  • Peristaltic pump with peristaltic 6‐mm tubing
  • 21‐G cannula
  • Metal tray
  • Absorbent tissue paper
  • Dissection instruments

Support Protocol 1: Fixation for Standard Histological Staining or Immunohistochemistry

  Materials
  • Excised lung tissue
  • Fixative: 10% NBF (see recipe) or 4% (w/v) paraformaldehyde (see recipe)
  • Filter paper or tissue paper
  • 70% (v/v) ethanol

Basic Protocol 2: Fixation of Nasal Cavity

  Materials
  • Mouse
  • Dissection instruments
  • Fixative: 10% NBF (see recipe) or 4% paraformaldehyde (see recipe)
  • Kristenson's fluid (see recipe)

Basic Protocol 3: Sampling Respiratory Tract Tissues for Histology

  Materials
  • Fixed and decalcified mouse head (see Basic Protocol 2)
  • Scalpel blade
  • Razor blade/microtome blade
  • Cutting board
  • Tissue cassettes
  • Downdraft table
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Figures

Videos

Literature Cited

Literature Cited
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  Scudamore, C.L. 2014. Practical approaches to reviewing and recording pathology data. In A Practical Guide to the Histology of the Mouse (C.L. Scudamore, ed.) pp. 25‐40. Wiley‐Blackwell, Hoboken, N.J.
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Key References
  McInnes, 2014. See above.
  Provides an overview of the basics of tissue collection and histology in the mouse, with information that is complementary to these protocols.
  Suarez et al., 2012. See above.
  Provides useful information on the macroscopic and microscopic comparative anatomy of humans and mice.
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