CYTOPLASMIC MICROTUBULES IN 2 DIFFERENT MOUSE MELANOMA CELL-LINES - AQUALITATIVE AND QUANTITATIVE-ANALYSIS USING CONFOCAL LASER-SCANNING MICROSCOPY AND COMPUTER-ASSISTED IMAGE-ANALYSIS

The microtubular system as one part of the cellular cytoskeleton is no t only necessary for mitotic activity of malignant cells but also for invading neighboring tissues and for the formation of distant metastas es. In the present study, the amount and distribution of tubulin in tw o murine melanoma cell lines (K1735-M2: high metastatic clone; K1735-c 116: low metastatic clone) were determined quantitatively using an ind irect immunofluorescence technique, confocal laser scanning microscopy (CLSM)and computer-assisted image analysis. Additionally, qualitative and quantitative changes after application of the microtubule-inhibit or nocodazole were investigated. Quantitative analysis showed a signif icant difference between the high and low metastatic cell line for the parameter TEXTURE, indicating a finer structured network within the h igh metastatic cells. After treatment with nocodazole the parameters T EXTURE and DENSITY were reduced, suggesting a decrease of assembled tu bulin and a less delicate structure of the remaining microtubules. Our study shows that CLSM combined with computer-assisted image analysis provides a new method to examine quantitative variations of the cytosk eleton possibly related to cell function. (C) Munksgaard 1997.