Automated Reconstruction Algorithm for Identification of 3D Architectures of Cribriform Ductal Carcinoma In Situ

Abstract

Ductal carcinoma in situ (DCIS) is a pre-invasive carcinoma of the breast that exhibits several distinct morphologies but the link between morphology and patient outcome is not clear. We hypothesize that different mechanisms of growth may still result in similar 2D morphologies, which may look different in 3D. To elucidate the connection between growth and 3D morphology, we reconstruct the 3D architecture of cribriform DCIS from resected patient material. We produce a fully automated algorithm that aligns, segments, and reconstructs 3D architectures from microscopy images of 2D serial sections from human specimens. The alignment algorithm is based on normalized cross correlation, the segmentation algorithm uses histogram equilization, Otsu's thresholding, and morphology techniques to segment the duct and cribra. The reconstruction method combines these images in 3D. We show that two distinct 3D architectures are indeed found in samples whose 2D histological sections are similarly identified as cribriform DCIS. These differences in architecture support the hypothesis that luminal spaces may form due to different mechanisms, either isolated cell death or merging fronds, leading to the different architectures. We find that out of 15 samples, 6 were found to have ‘bubble-like’ cribra, 6 were found to have ‘tube-like’ criba and 3 were ‘unknown.’ We propose that the 3D architectures found, ‘bubbles’ and ‘tubes’, account for some of the heterogeneity of the disease and may be prognostic indicators of different patient outcomes.

Three‐dimensional microscopic analysis of clinical prostate specimens

“…Three‐dimensional microscopic tissue reconstruction has mostly been carried out by serial sectioning, staining and imaging of up to hundreds of 2D sections . Although developments in image stacking have improved 3D reconstruction, serial slicing is laborious and sensitive to tissue deformations . Confocal laser scanning microscopy (CLSM) allows fluorescent 3D imaging without the need for sectioning, but is limited to depths of tens of micrometres, owing to scattering and absorption of emitted fluorescent light.…”

Three‐dimensional reconstruction of ductal carcinoma in situ with virtual slides

“…During this study, it was noted that fine detail included in the segmentation may be unnecessary, thus reducing the resolution the time taken doing this task can be reduced. The accuracy of segmentation was discussed by Norton, who stated that 90% accuracy was not necessary to determine the 3D structure. Norton developed an algorithm that enabled successful automatic segmentation of cribra of DCIS, analogous to the lumina segmented in our study.…”

“…Today the tools available to study 3D histology have improved substantially such that high resolution archivable digital images may permit more elaborate studies. It is now possible to generate image stacks of serially sectioned tissue and most recently, computer‐generated 3D reconstructed graphical models of breast carcinoma . Such reconstructions have demonstrated differences between tumours that appear histologically similar in 2D .…”

Development of a 2D Image Reconstruction and Viewing System for Histological Images from Multiple Tissue Blocks: Towards High-Resolution Whole-Organ 3D Histological Images

“…Whole-slide imaging (WSI) technology, which enabled us to capture the high-resolution images of stained tissue sections, has not only encouraged the development of various automated histopathology image analysis systems for diagnostic supports [4,5,6,7,8,9,10,11,12], the technology has also spurred interest in 3D histology image reconstruction of tissue organs [13,14,15,16,17]. Reconstruction of the 3D volumetric image of a tissue organ from series of histology images is an emerging application area in digital pathology.…”

“…Initial works on high-resolution 3D histology image reconstruction considered a single formalin-fixed paraffin-embedded (FFPE) tissue block [13,14,15,16,17]. Series of thin tissue sections were cut from the FFPE tissue blocks, mounted on standard-size glass slides and then stained.…”