Standarisasi Analisis Histomorfometri Kerapatan Kolagen Menggunakan ImageJ sebagai Upaya Peningkatan Mutu Penelitian Mahasiswa di Laboratorium Biomedik Kedokteran Gigi

Histomorphometric analysis of collagen density is widely used in dental biomedical research; however, visual assessment is susceptible to interobserver variation. This study developed, implemented, and evaluated a standardized ImageJ based protocol for collagen density histomorphometry in a dental biomedical laboratory. This descriptive quantitative implementation study comprised standard operating procedure (SOP) development, observer training, and implementation using Masson’s Trichrome-stained histological specimens. Images were calibrated, processed by color deconvolution and thresholding, and measured as collagen area fraction. Interobserver reliability was evaluated using the intraclass correlation coefficient (ICC). The SOP was implemented in all specimens. The mean collagen density was 38.87 ±3.59%, with an ICC of 0.87 (95% confidence interval: 0.79-0.93), indicating good interobserver reliability. The mean analysis time decreased from 15-20 minutes with the manual method to 3-5 minutes using ImageJ. ImageJ-based analysis and the SOP were adopted in 85% and 83% of student research projects, respectively. The standardized ImageJ based protocol demonstrated good reliability, improved analytical efficiency, and supported laboratory quality assurance. It is applicable for standardized histological analysis in dental biomedical laboratory education.