189 Pattern recognition in pulmonary AdC; a modified classification Furthermore, the presence of a scar, defined as a (peri)centrally in the tumour located, recognizable area of fibrosis and/or collapse with elastosis was estimated in each tumour. The composition of the scar was scored as only fibrotic, only biological collapse or the combination of both. In each case the presence of invasive growth into the scar area was scored as absent, extensively present, focal in the fibrotic or biological collapse part or in both. The immunohistochemical stain for Cytokeratin 7 (CK7) (clone OVTL12/30 (Agilent/ Dako (Glostrup, Denmark, catno. M701801) was performed in a Roche/Ventana benchmark Ultra (Roche, Basel Switzerland) with 4 µm tissue slides mounted on TOMOglass slides (Roche, Basel, Switzerland). Antigen retrieval was applied with high pHbuffer CC1(32 minutes at 100°C) and the antibody was diluted 1/100 (incubated for 32 minutes at 36°C) and detected with the Optiview DAB kit under standard conditions. Sections were dehydrated with ethanol 100%, cleared with Xylene and coverslipped with Tissue-Tek coverslip film on the Sakura coverslipper (Sakura Finetek Europe B.V, Alphen aan den Rijn, The Netherlands). Internal positive controls were pneumocytes and respiratory epithelial cells. Internal negative controls were e.g., smooth muscle cells in vessel walls and bronchioli. Statistical analysis In the morphometrical part, Pearson’s correlation coefficients were estimated between amount of air dots and tumour cell height. In the 2 cohorts, survival times, measured as a) recurrence free survival (RFS, time from initial surgery to recurrence); b) overall survival (OS, time from surgery to death from any cause)331, were compared between cases identified as collapsed AIS and invasive cases using the log rank test. Survival times are plotted with Kaplan-Meier curves, 5-year survival were reported with corresponding 95% confidence interval (CI). The significance level was set at 0.05. All analyses were conducted in SPSS version 26 (IBM Corp., Armonk, NY). Results Morphological consequences of the mathematical model In the mathematical model the 3-dimensional configuration, comparable to alveolar structures was used. A graphical display with a 2-dimensional and a 3-dimensional approach is shown in Figure 10. 14
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