Combined serial section‐based 3D reconstruction of cervical carcinoma invasion using H&E/p16^INK4a/CD3 alternate staining

Abstract

Background: Malignant growth and invasiveness of cancers is a function of both intratumoral and stromal factors. The accessibility to nutrients, oxygen and growth factors, the stromal composition, and the interference with the immune system all shape the tumor invasion front. A recent study has shown a prognostic difference with respect to different invasion patterns analyzed on histological specimens of cervical cancers. The present study analyzes the spatial organization of a cervical cancer and the relation of the tumor invasion front and the infiltration with CD31 T-cells. Methods: From a cervical squamous cell carcinoma specimen, 84 serial sections were performed and three interleaving series were stained with hematoxylin/eosin and immunohistochemistry directed against the cervical carcinoma biomarker p16INK4a and the T-cell marker CD3. Sections were passed through an image processing chain to obtain a reconstructed and segmented tissue volume. For local tumor invasion front analysis the mean curvature was used, which in turn was related to the respective local minimum tumor to T-cell distance as well to a T-cell originated diffusing substance's concentration at the tumor surface.

Image‐processing chain for a three‐dimensional reconstruction of basal cell carcinomas*†

“…At present, there is still no 3D imaging modality providing both an appropriate field of view and a sufficient penetration depth at microscopic resolution. For specimen from other cancer types, a large histological serial section‐based 3D reconstruction framework has already been established (4,5). This work provides proof of principle for a novel automated image‐processing chain for serial section‐based 3D BCC reconstruction capable to address the diffuse tumor structures of that type of skin cancer.…”

Three‐dimensional reconstruction with serial whole‐mount sections of oral tongue squamous cell carcinoma: A preliminary study

“…As a novel emerging method, histopathological 3D reconstruction has been applied in the study of cell‐cell interaction in cancer, cancer cell invasion, and EMT marker, as well as vessel study . As for 3D observation of OTSCC, previous study on invasion front of OTSCC has been limited using limited parts of OTSCC samples, or only microinvasive OTSCC samples .…”

“…Each piece was sliced into approximately 700 4‐μm sections with horizontal sliding microtome (Rem‐710; YAMATO, Tokyo, Japan). Combined staining was performed as previously described . Briefly, each section stack was separated into 3 interleaving series, each with approximately 233 sections.…”

Three-dimensional visualization and quantification for the growth and invasion of oral squamous cell carcinoma

“…Recent advance in 3D imaging technology allows us to inspect the details of tumor architecture [12][13][14][15]. In the past few years, we initiated to reconstruct SCC architectures at the invasion front using serial histological sections and, on that basis, to document growth and invasion patterns of SCC.…”