TY  - JOUR
TI  - Visualizing the Osteocyte Lacuno-Canalicular System via a Rapid 10-Minute Silver Nitrate Staining Method
AU  - Wu, Jinlian 
AU  - Dai, Weiwei 
AU  - Wang, Libo 
VL  - 16
IS  - 15
PY  - 2026
DA  - 2026/08/05
SP  - e5784
C1  - Bio-protocol 2026;16:e5784
DO  - 10.21769/BioProtoc.5784
UR  - https://doi.org/10.21769/BioProtoc.5784
AB  - The conventional Ploton silver method employs a high-concentration 50% (w/v; 2.943 mol/L) silver nitrate solution for histological staining and characterization of the osteocyte lacuno-canalicular system (LCS). However, it is limited by prolonged staining times (55 min) and by risks of LCS ultrastructural damage and/or incomplete impregnation. To address these limitations, we developed the Wu–Wang silver nitrate staining method, which uses a 1 mol/L silver nitrate solution under elevated temperature (50–70 °C) to achieve rapid, effective, and high-contrast visualization of the osteocyte LCS within 10 min. We further demonstrate that this novel method enables robust LCS visualization across multiple vertebrate species. Compared with the Ploton method, the Wu–Wang method substantially reduces staining time and overcomes staining limitations inherent to prolonged exposure to concentrated silver nitrate solutions. This rapid and efficient staining method supports more accurate quantitative analysis of LCS morphology and facilitates systematic investigation of osteocyte and LCS morphogenesis, as well as the pathological mechanisms underlying bone and joint disease.
KW  - Wu–Wang silver nitrate staining method
KW  - Ploton silver nitrate staining method
KW  - Osteocyte
KW  - Lacuno-canalicular system (LCS)
KW  - Bone
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False