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

Abstract

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.

Studying Wound Repair in the Mouse

“…no. E105 and E102) Coverslips, thickness 1 (Thermo Scientific, D10143263NR) Bright-field microscope connected to color digital camera Additional reagents and equipment for euthanasia of mice (Donovan and Brown, 2006b), fixation of tissue (Support Protocol 3, steps 5 to 8), and sectioning of tissue (Antal et al, 2011) Isolation of wound biopsies 1. Euthanize the mouse (Donovan and Brown, 2006b) according to locally approved protocol.…”

“…3. Fix (Support Protocol 3, steps 5 to 8) and section (see, e.g., Antal et al, 2011) the wound biopsies according to standard protocol. Place the sections on SuperFrost Plus microscope slides.…”

“…1. Subject the wound sections to different staining techniques, such as hematoxylineosin or Masson trichrome stains (also see Antal et al, 2011).…”

Labeling and Magnetic Resonance Imaging of Exosomes Isolated from Adipose Stem Cells

“…Our Strategic Planning is described schematically in the Figure 3.44.1. Refrigerated centrifuge 70-μm nylon mesh cell strainer 75-cm 2 cell culture flasks (BD Falcon) 15-and 50-ml conical polypropylene centrifuge tubes (e.g., Corning Falcon) Hemacytometer (Bürker chamber; see UNIT 1.1; Phelan & May, 2015) Light microscope (Olympus BXS1) UV-vis spectrophotometer MRI tomograph at 4.7 T (e.g., Bruker Tomograph equipped with 4.7-T, 33-cm-bore horizontal magnet) Heated bed for mouse Microtome equipped with diamond knife Cu/Rh grids (Electron Microscopy Sciences) Transmission electron microscope (TEM; Philips Morgagni) equipped with digital camera Additional reagents and equipment for dissection of the mouse (Antal, Muller, Wendling, Hérault, & Mark, 2011;Treuting & Snyder, 2015), basic cell culture techniques including growth, trypsinization, and counting of cells (UNIT 1.1; Phelan & May, 2015), anesthesia of mice (Donovan & Brown, 1998), and injection of mice (Donovan & Brown, 2006)…”

“…1. Isolate inguinal adipose tissues from C57BL/6 mice (Antal, Muller, Wendling, Hérault, & Mark, 2011;Treuting & Snyder, 2015).…”