Abstract

Background: The von Kossa staining technique is an important method in pathology and histology, used to detect the presence of calcium in body tissues. This technique supports the diagnosis of several pathological conditions, including Malakoplakia - a rare inflammatory disease that may be confused with other conditions. Standardizing the von Kossa staining protocol is therefore beneficial for practical application. Objective: To standardize an effective von Kossa staining protocol. Materials and methods: This is a descriptive experimental study performed on the confirmed case of Malakoplakia. Tissue samples were sectioned at thicknesses of 3, 4, 5, and 6 µm, with incubation in 5% silver nitrate for 15, 30, 45, and 60 minutes, respectively. Each experiment was repeated three times and staining effectiveness was evaluated based on the demonstration of Michaelis-Gutmann bodies. Results: The staining method highlighted Michaelis-Gutmann bodies as black “target-like” structures. The results depended on both section thickness and incubation time in 5% silver nitrate. With a short incubation time of 15 minutes, most samples showed light brown to dark brown staining and did not achieve the required contrast. At 30 - 45 minutes, the M-G bodies in 4 µm and 5 µm sections showed the characteristic target-like appearance with uniform black staining. After 60 minutes of incubation, all sections stained intensely black; however, overstaining occurred in thinner sections, obscuring morphological details. Conclusion: The standardized protocol recommends a section thickness of 4 µm and incubation in 5% silver nitrate for approximately 45 minutes to achieve the best staining results.