[Staining of bone canaliculi using decalcified bone tissues].
Recently we were able to stain bone canaliculi using the decalcified bone tissues, according to a modified method of the Bodian staining method, which is commonly used for staining the nerve fibers with silver impregnation of protargol. The bone tissues from normal rabbits, rats, and humans were decalcified with buffered EDTA solution after 10% formalin fixation. The tissues were embedded in paraffin and were sliced at 4 microns then the sections were attached to the gelatin-coated glass slides. These sections were treated first with potassium dichromate followed by 2% protargol solution containing copper. After gold chrolide treatment, they were immersed in oxalic acid. Both the lacunae and canaliculi of osteocytes were stained black in sharp contrast to bone matrix of red purple. Using the present staining method, the bone canaliculi were clearly detectable in fine details without producing apparent artificial damage of soft tissues. The canaliculi showed also markedly complex fine structures. By combining the present staining method with eosin or toluidine blue staining, we were able to recognize even immature osteocytes in enchondral ossification at growth cartilage with fine bone canaliculi. From these results, we conclude that the present method is very useful in histopathological studies of the lacuna-canalicular system of osteocytes in the decalcified bone tissues.
Kageyama
N
N
Department of Orthopaedic Surgery, Kyoto Prefectural University of Medicine, Japan.
Kusuzaki
K
K
Yamashita
F
F
Hirasawa
Y
Y
Ashihara
T
T
D000818:Animals; D001842:Bone and Bones / Q000166:cytology / Q000648:ultrastructure*; D006801:Humans; D011817:Rabbits; D051381:Rats; D013194:Staining and Labeling / Q000379:methods*