Several cellular mechanisms affect nuclear morphology which can therefore be used to assess certain processes. Here, we present an analytic tool to quantify the number of cells in a population that present characteristics of senescence, apoptosis or nuclear irregularities through nuclear morphometric analysis. The tool presented here is based on nuclear image analysis and evaluation of size and regularity of adhered cells in culture. From 46 measurements of nuclear morphometry, principal component analysis filtered four measurements that best separated regular from irregular nuclei. These measurements, namely aspect, area box, radius ratio and roundness were combined into a single nuclear irregularity index (NII). Normal nuclei are used to set the parameters for a given cell type, and different nuclear phenotypes are separated in an area versus NII plot. The tool was validated with β-gal staining for senescence and annexin or caspases inhibitor for apoptosis as well as several treatments that induce different cellular phenotypes. This method provides a direct and objective way of screening normal, senescent, apoptotic and nuclear irregularities which may occur during failed mitosis or mitotic catastrophe, which may be very useful in basic and clinical research.

The authors present this study using sodium fluorescein (SF) to enhance skull base tumors by performing a quantitative digital analysis of tumor enhancement. The purpose of this study is to observe the grade of SF enhancement by the tumors. A prospective experiment within-subjects study design was performed which included six patients with skull base lesions. Digital pictures were taken before and after the SF systemic injection, using the same light source of the microsurgical field. The pictures were analyzed by computer software which calculated the wavelength (WL) of the SF pre- and post-injection. THE GROUP OF TUMORS WAS AS FOLLOWS: one vestibular schwannoma, three meningiomas, one craniopharyngioma and one pituitary adenoma. The SF enhancement in all tumors was strongly positive. The digital analysis of the pictures, considering the SF WL pre- and post-injection, presented P = 0.028 (Wilcoxon T test). The enhancement of the tumors by SF was consistent and evident. The introductory results suggest the possibility of using SF as an adjuvant tool for the skull base surgery. Further studies should test the clinical application of the SF in skull base tumors.