Affiliations: Institute of Oncology, 1000 Ljubljana, Slovenia | Faculty of Electrical Engeneering, 1000 Ljubljana, Slovenia
Note: [] Correspondence to: Margareta Fležar, MD, Dept. Cytopathology, Institute of Oncology, Zaloška 2, 1105 Ljubljana, Slovenia. Tel.: +386 61 132 20 99; Fax: +386 61 1314 180; E‐mail: mflezar@onko‐i.si.
Abstract: The aim of the study was to determine optimal hydrolysis time for the Feulgen DNA staining of archival formalin fixed paraffin‐embedded surgical samples, prepared as single cell suspensions for image cytometric measurements. The nuclear texture features along with the IOD (integrated optical density) of the tumor nuclei were analysed by an automated high resolution image cytometer as a function of duration of hydrolysis treatment (in 5 N HCl at room temperature). Tissue blocks of breast carcinoma, ovarian serous carcinoma, ovarian serous tumor of borderline malignancy and leiomyosarcoma were included in the study. IOD hydrolysis profiles showed plateau between 30 and 60 min in the breast carcinoma and leiomyosarcoma, and between 40 and 60 min in the ovarian serous carcinoma and ovarian serous tumor of borderline malignancy. Most of the nuclear texture features remained stable after 20 min of hydrolysis treatment. Our results indicate that the optimal hydrolysis time for IOD and for nuclear texture feature measurements, was between 40 and 60 min in the cell preparations from tissue blocks of three epithelial and one soft tissue tumor.
