Abstract
The objective of the present study was to establish a histology-based 3D reconstruction procedure and thus to improve our understanding of the tumor microenvironment. Based on a pathodiagnostic survey, tissue specimens (3 mm in diameter) were dissected out from the invasion front of tongue squamous cell carcinoma (SCC) using a paraffin tissue microarray. The experimental protocol comprised step-wise immunolabeling of serially cut sections (4 μm thick) and image acquisition using a Virtual Microscopy system (Nano-Zoomer, Hamamatsu) that certifies the quality of the digitized images, particularly their spatial and RGB color resolution. Image registration and segmentation for 3D reconstruction were automated with the aid of Ratoc TRI-SRF2 software to avoid operator-dependent subjectivity. Visualization and quantification of 3D constructs facilitated validation of the continuity or segregation of carcinoma cells in the circumscribing stroma and assessment of the frequency of spatial contact between cancer cells and blood/lymph vessels. These 3D parameters are of great value for the prediction and diagnosis of cancer aggressiveness and malignancy in the coherent microenvironment. The results of this study demonstrate the advantages and suggest the wide range of potential applications of histology-based 3D reconstruction for evaluating biological tissue architecture and the interaction of multiple constituents.
