PARAMETERS DERIVED FROM INTEGRATED NUCLEAR FLUORESCENCE, SYNTACTIC STRUCTURE-ANALYSIS, AND VASCULARIZATION IN HUMAN LUNG CARCINOMAS

Combined measurements of integrated nuclear fluorescence (INF) and vas cularization were performed on surgical specimens of human lung carcin omas. Histological slides of formalin-fixed, paraffin-embedded tissue samples were treated with Texas Red-labeled antibody to factor Vm and the fluorochrome DAPI. The resulting images were analyzed with an epii llumination fluorescence microscope and two different filter blocks. T he first image displayed the vessels, and the second the DAPI-stained nuclei of surrounding cells. The extent of vascularization was assesse d by calculating the volume fraction (Vv), the surface fraction (Sv), the area, and the minimum diameter of the vessels. The INF was measure d in tumour cells and lymphocytes, and was grouped according to the di stance from the nearest vascular boundary into the intervals of 0-20, 21-40, 41-60, 61-80, and >80 mu. The numerical densities (Nv) as well as the percentages of S-phase-related tumour cell fraction (SPRF) and of tumour cells with an INF > 5C were computed. A minimum of 50 vessel s and 300 tumour cells were examined. The material included 100 cases with primary lung carcinoma (39 epidermoid carcinomas, 39 adenocarcino mas, 13 large cell carcinomas, three small cell anaplastic carcinomas, and 6 carcinoid tumours). On the average, the volume density of the s troma amounts to 16.7%, and that of the vessels (Vv) to 12.8%. The min imum diameter of the intratumoral vessels is 13 mu and the measured ci rcumference 138 mu. The numerical densities of tumour cells (lymphocyt es) decrease with increasing distance from the vascular boundary from 6.3 (1.7) to 1.0 (0.1). A reduction is also seen in the percentage of the SPRF from 10.7 to 8.1%. The percentage of tumour cells with an INF > 5C, however, is positively correlated to the distance from the vasc ular surfaces from 34.2 to 38.2%. The measurements reveal that tumour cells are densely positioned and have an increased proportion of proli feration in the populations close to perivascular spaces, whereas chro mosome abnormalities are seen more frequently, when tumour cells are l ocated at a distance >20 mu from the vascular surfaces.