Tips and techniques for processing and sectioning undecalcified murine bone specimens.
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.
Indiana University School of Medicine, Indianapolis, IN, USA.
Thomas B
TB
Bemenderfer
Indiana University School of Medicine, Indianapolis, IN, USA.
Jonathan S
JS
Harris
Indiana University School of Medicine, Indianapolis, IN, USA.
Keith W
KW
Condon
Indiana University School of Medicine, Indianapolis, IN, USA.
Melissa A
MA
Kacena
D000818:Animals; D001842:Bone and Bones / Q000166:cytology* / Q000378:metabolism*; D002113:Calcification, Physiologic; D016591:Histocytological Preparation Techniques; D051379:Mice; D008867:Microtomy / Q000295:instrumentation / Q000379:methods; D013194:Staining and Labeling / Q000379:methods