TY  - JOUR
TI  - A Novel Optimized Silver Nitrate Staining Method for Visualizing and Quantifying the Osteocyte Lacuno-Canalicular System (LCS)
AU  - Wu, Jinlian 
AU  - Xue, Chunchun 
AU  - Li, Qiang 
AU  - Wu, Hongjin 
AU  - Zhang, Jie 
AU  - Wang, Chenglong 
AU  - Dai, Weiwei 
AU  - Wang, Libo 
VL  - 15
IS  - 8
PY  - 2025
DA  - 2025/04/20
SP  - e5289
C1  - Bio-protocol 2025;15:e5289
DO  - 10.21769/BioProtoc.5289
UR  - https://doi.org/10.21769/BioProtoc.5289
AB  - The osteocyte lacuno-canalicular system (LCS) plays a crucial role in maintaining bone homeostasis and mediating cellular mechanotransduction. Current histological techniques, particularly the Ploton silver nitrate staining method, face challenges such as variations in solution concentrations and types as well as a lack of standardization, which limits their broader application in osteocyte research. In this study, we present a simplified and more effective silver nitrate staining protocol designed to address these issues. Our method utilizes a 1 mol/L silver nitrate solution combined with optimized gelatin-formic acid solutions at varying concentrations (0.05%–0.5% type-B gelatin and 0.05%–5% formic acid, or 1%–2% type-B gelatin and 0.1%–2% formic acid). Staining is performed for 1 h under 254 nm ultraviolet light or 90 min under room light, followed by washing with Milli-Q water to terminate staining. This novel optimized method yields consistent and distinct staining of the osteocyte LCS across multiple species, demonstrating superior efficiency and reliability compared to the Ploton method. It will significantly advance research in osteocyte biology and provide a valuable tool for exploring the adaptive evolution of osteocyte LCS morphology and function across various taxa.
KW  - Silver Nitrate
KW  - Gelatin
KW  - Formic acid
KW  - Osteocyte
KW  - Lacuno-canalicular System (LCS)
KW  - Bone
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False