Tips and techniques for processing and sectioning undecalcified murine bone specimens

Thomas B. Bemenderfer, Jonathan S. Harris, Keith W. Condon, Melissa A. Kacena

Research output: Chapter in Book/Report/Conference proceedingChapter

9 Scopus citations


Preparation of mineralized tissue specimens for bone-specific staining encompasses a critical sequence of histological techniques that provides visualization of tissue and cellular morphology. Bone specimens are fixed in 10 % neutral-buffered formalin, dehydrated in graded ethanol (EtOH) solutions (and optionally cleared in xylene), infiltrated and embedded in polymethyl methacrylate (methyl methacrylate), classically sliced into 4-10 micrometer (μm) sections, and stained with bone-specific histological stains such as von Kossa (with either nuclear fast red solution counterstain or MacNeal's tetrachrome counterstain), modified Goldner's trichrome, and alizarin red S stain. Here, we describe the tissue processing of mineralized mouse bones from dissection to staining for histological analysis by light microscopy.

Original languageEnglish (US)
Title of host publicationSkeletal Development and Repair
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Number of pages25
ISBN (Print)9781627039888
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Bone
  • Dehydration
  • Embedding
  • Fixation
  • Methyl methacrylate
  • Sectioning
  • Staining

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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