Safranin-O Staining: A Method to Visualize the Structural Integrity and Cellularity of Cartilage

Abstract

Source: Zhao, Y., et al. Establishment of a Surgically-induced Model in Mice to Investigate the Protective Role of Progranulin in Osteoarthritis. J. Vis. Exp. (2014).

In this video, we demonstrate a staining procedure for cartilage using safranin-orange. This method helps visualize the cartilage and detect any damage.

Protocol

1. Safranin O staining

1. Begin with the glass slides containing serial sagittal sections spanning a region from the center of the lateral condyle to the center of the medial condyle of a euthanized mouse model with the destabilized medial meniscus.
2. Next, deparaffinize the slides in oxylene, and hydrate them in an ethanol gradient to distilled water. Thereafter, the slides are stained through Weigert's Iron Hematoxylin Solution, 0.05% Fast Green (FCF) Solution, 1% Acetic Acid Solution, and 0.1% Safranin O Solution. Mount the slides using resinous medium.
3. Score the Safranin O-stained slides based on loss of proteoglycan (red color) in cartilage and percentage of destruction in cartilage structure, do statistical analysis for the histological score.