Computer-assisted image analysis in the diagnosis of gynaecological lesions: A quantitative and comparative investigation of haematoxylin-eosin with special dyes on tissue

Morphometry and quantitative image analysis have shown considerable potential for diagnostic applications because they eliminate subjectivity and increase the measurement reproducibility. In our study, we investigated the use of morphometric quantitative image analysis as a tool to aid diagnosis of lesion tissues stained with seven staining techniques. Thirteen already established lesions were examined in this study. Tissues were sectioned at 5 μm thickness using the rotary microtome and prepared for staining according to standard methods before staining them with seven different histochemical dyes: Haematoxylin and Eosin (H&E), Masson's Trichrome (MT), Periodic Acid Schiff (PAS), Phosphotungstic Acid Haematoxylin (PTAH), Southgate Mucincamine (SGM), Alcian Blue (AB) and Verhoeff Van Gieson (VVG). Photomicrographs were analyzed using imageJ for percentage area and intensity measurement. The average % area and intensity measurement based on the seven staining techniques used in this study were 47.88% and 122.23 points (pts). The lesions, adenomyosis in MT (80%), squamous cell carcinoma in PAS (80%) and fibroma in PTAH (77%) stained above the average % area. The squamous cell carcinoma stained with MT was the most intense tissue (55 pts), followed by Brenner tumour in PTAH (61 pts) and squamous cell carcinoma in PAS (69 pts). There were significant correlations between the % area of tissues covered by PAS and H&E (r = 0.599, p = 0.030), and PAS and MT (r = 0.572, p = 0.041) as well as in the intensity of tissues in PAS and H&E (r=0.615, p=0.025), VVG and H&E (r = 0.707, p = 0.007), and VVG and PTAH (r = 0.577, p = 0.039). MT staining technique produced the best results in the % area of tissues covered and the intensity measurements, and therefore was recommended for routine use alongside with H&E in diagnostic histopathology.