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Novel polychrome staining distinguishing osteochondral tissue and bone cells in decalcified paraffin sections

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Title: Novel polychrome staining distinguishing osteochondral tissue and bone cells in decalcified paraffin sections
Authors: Nakamura, Teppei Browse this author →KAKEN DB
Sumi, Kanako Browse this author
Tsuji, Erika Browse this author
Hosotani, Marina Browse this author
Namba, Takashi Browse this author
Ichii, Osamu Browse this author →KAKEN DB
Irie, Takao Browse this author
Nagasaki, Ken-ichi Browse this author
Kon, Yasuhiro Browse this author →KAKEN DB
Mishima, Takashi Browse this author
Yoshiyasu, Tomoji Browse this author
Keywords: Osteoid
Mineralized bone
Bone cells
Osteochondral staining
Issue Date: 19-Aug-2021
Publisher: Springer
Journal Title: Cell and tissue research
Volume: 385
Start Page: 727
Publisher DOI: 10.1007/s00441-021-03516-6
Abstract: The bone is a dynamic and metabolically active organ in which growth and resorption of the osteochondral matrix is orchestrated by osteoblasts and osteoclasts. For decalcified paraffin-embedded specimens, decalcifying agents alter the staining intensity, and excess decalcification interferes with bone staining. Robust bone staining methods independent of the decalcification conditions and animal species are lacking. In this study, we have developed a novel polychrome staining method, named JFRL staining, which stains the components of osteochondral tissue in different colors. With this staining we could visualize the hyaline cartilage as blue by alcian blue, osteoid as red by picrosirius red, and mineralized bone as green by picro-light green SF or picro-naphthol green B and easily distinguished osteoblasts, osteocytes, and osteoclasts. In mineralized bone, this staining revealed the obvious lamellar structures and woven bone. Notably, this staining was independent of the decalcification conditions and experimental animal species examined. To verify the usefulness of JFRL staining, we observed cotton rat tail which has shorter length and shows a false autotomy. The caudal vertebrae were normally developed via endochondral ossification without a fracture plane. At 6 months of age, the number of chondrocytes declined and the hypertrophic zone was absent at the epiphyseal plate, which might reflect the shorter tail. In conclusion, JFRL staining is the first method to simultaneously distinguish osteochondral matrix and bone cells in one section regardless of decalcifying conditions. This robust staining will provide new information for a wide number of biomedical fields, including bone development, physiology, and pathology.
Rights: This is a post-peer-review, pre-copyedit version of an article published in Cell and tissue research. The final authenticated version is available online at:
Type: article (author version)
Appears in Collections:獣医学院・獣医学研究院 (Graduate School of Veterinary Medicine / Faculty of Veterinary Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 中村 鉄平

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