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Three-dimensional reconstruction of oral tongue squamous cell carcinoma at invasion front.

Kudo T, Shimazu Y, Yagishita H, Izumo T, Soeno Y, Sato K, Taya Y, Aoba T - Int J Dent (2013)

Bottom Line: Serial sections (4  μ m thick) were double immunostained with pan-cytokeratin and Ki67 antibodies and digitized images were acquired using virtual microscopy.Direct visualization and quantitative assessment of the parenchymal-stromal border provide a new dimension in our understanding of OTSCC architecture.These 3D morphometric analyses also ascertained that cell invasion (individually and collectively) occurs at the deep invasive front of the OTSCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan ; Department of Pathology, School of Life Dentistry at Tokyo, The Nippon Dental University, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan.

ABSTRACT
We conducted three-dimensional (3D) reconstruction of oral tongue squamous cell carcinoma (OTSCC) using serial histological sections to visualize the architecture of invasive tumors. Fourteen OTSCC cases were collected from archival paraffin-embedded specimens. Based on a pathodiagnostic survey of whole cancer lesions, a core tissue specimen (3 mm in diameter) was dissected out from the deep invasion front using a paraffin tissue microarray. Serial sections (4  μ m thick) were double immunostained with pan-cytokeratin and Ki67 antibodies and digitized images were acquired using virtual microscopy. For 3D reconstruction, image registration and RGB color segmentation were automated using ImageJ software to avoid operator-dependent subjective errors. Based on the 3D tumor architecture, we classified the mode of invasion into four types: pushing and bulky architecture; trabecular architecture; diffuse spreading; and special forms. Direct visualization and quantitative assessment of the parenchymal-stromal border provide a new dimension in our understanding of OTSCC architecture. These 3D morphometric analyses also ascertained that cell invasion (individually and collectively) occurs at the deep invasive front of the OTSCC. These results demonstrate the advantages of histology-based 3D reconstruction for evaluating tumor architecture and its potential for a wide range of applications.

No MeSH data available.


Related in: MedlinePlus

Four types of tumor architecture and mitotic activity at the OTSCC invasion front. Images are from left to right: the immunostained microscopic (2D) image; 3D view of the segmented tumor parenchyma; 3D view of infiltrating cancer foci (yellow) detached in all dimensions from the bulk tumor parenchyma (gray); and Ki67-positive nuclei in the tumor parenchyma (red) and stroma (blue). Additional 3D data obtained from the remaining 10 OTSCC cases can be found in Supplementary Material Plates S2 and S3. Bar = 1 mm.
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fig4: Four types of tumor architecture and mitotic activity at the OTSCC invasion front. Images are from left to right: the immunostained microscopic (2D) image; 3D view of the segmented tumor parenchyma; 3D view of infiltrating cancer foci (yellow) detached in all dimensions from the bulk tumor parenchyma (gray); and Ki67-positive nuclei in the tumor parenchyma (red) and stroma (blue). Additional 3D data obtained from the remaining 10 OTSCC cases can be found in Supplementary Material Plates S2 and S3. Bar = 1 mm.

Mentions: Figure 4 compares microscopic (2D) and reconstructed (3D) images of tumor architecture and invasion mode in four cases of OTSCC; the remaining data from the other OTSCC cases are presented in Supplementary Material Plates S2 and S3. Using the CK cocktail to label cancer cells, 2D views reveal heterogeneity in the patterns of cancer cell invasion both between OTSCC cases and within individual cases. Among the infiltrating cancer cells observed on 2D images, the 3D segmentation protocol enabled the visualization of discrete cancer cells and clumps that were detached from each other and completely encased in the stroma. We herein use the term “discohesive cancer foci” to designate a pool of infiltrating cancer cells segmented in 3D space. We also addressed locoregional differences in mitotic activity as indicated by the density of Ki67-positive nuclei in the parenchymal and stromal segments.Based on the 3D features of tumor architecture, we discerned four types: (1) pushing and bulky architecture with short finger-like projections; (2) trabecular architecture with strands and cords; (3) diffuse spreading; and (4) special forms.


Three-dimensional reconstruction of oral tongue squamous cell carcinoma at invasion front.

Kudo T, Shimazu Y, Yagishita H, Izumo T, Soeno Y, Sato K, Taya Y, Aoba T - Int J Dent (2013)

Four types of tumor architecture and mitotic activity at the OTSCC invasion front. Images are from left to right: the immunostained microscopic (2D) image; 3D view of the segmented tumor parenchyma; 3D view of infiltrating cancer foci (yellow) detached in all dimensions from the bulk tumor parenchyma (gray); and Ki67-positive nuclei in the tumor parenchyma (red) and stroma (blue). Additional 3D data obtained from the remaining 10 OTSCC cases can be found in Supplementary Material Plates S2 and S3. Bar = 1 mm.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3818895&req=5

fig4: Four types of tumor architecture and mitotic activity at the OTSCC invasion front. Images are from left to right: the immunostained microscopic (2D) image; 3D view of the segmented tumor parenchyma; 3D view of infiltrating cancer foci (yellow) detached in all dimensions from the bulk tumor parenchyma (gray); and Ki67-positive nuclei in the tumor parenchyma (red) and stroma (blue). Additional 3D data obtained from the remaining 10 OTSCC cases can be found in Supplementary Material Plates S2 and S3. Bar = 1 mm.
Mentions: Figure 4 compares microscopic (2D) and reconstructed (3D) images of tumor architecture and invasion mode in four cases of OTSCC; the remaining data from the other OTSCC cases are presented in Supplementary Material Plates S2 and S3. Using the CK cocktail to label cancer cells, 2D views reveal heterogeneity in the patterns of cancer cell invasion both between OTSCC cases and within individual cases. Among the infiltrating cancer cells observed on 2D images, the 3D segmentation protocol enabled the visualization of discrete cancer cells and clumps that were detached from each other and completely encased in the stroma. We herein use the term “discohesive cancer foci” to designate a pool of infiltrating cancer cells segmented in 3D space. We also addressed locoregional differences in mitotic activity as indicated by the density of Ki67-positive nuclei in the parenchymal and stromal segments.Based on the 3D features of tumor architecture, we discerned four types: (1) pushing and bulky architecture with short finger-like projections; (2) trabecular architecture with strands and cords; (3) diffuse spreading; and (4) special forms.

Bottom Line: Serial sections (4  μ m thick) were double immunostained with pan-cytokeratin and Ki67 antibodies and digitized images were acquired using virtual microscopy.Direct visualization and quantitative assessment of the parenchymal-stromal border provide a new dimension in our understanding of OTSCC architecture.These 3D morphometric analyses also ascertained that cell invasion (individually and collectively) occurs at the deep invasive front of the OTSCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan ; Department of Pathology, School of Life Dentistry at Tokyo, The Nippon Dental University, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan.

ABSTRACT
We conducted three-dimensional (3D) reconstruction of oral tongue squamous cell carcinoma (OTSCC) using serial histological sections to visualize the architecture of invasive tumors. Fourteen OTSCC cases were collected from archival paraffin-embedded specimens. Based on a pathodiagnostic survey of whole cancer lesions, a core tissue specimen (3 mm in diameter) was dissected out from the deep invasion front using a paraffin tissue microarray. Serial sections (4  μ m thick) were double immunostained with pan-cytokeratin and Ki67 antibodies and digitized images were acquired using virtual microscopy. For 3D reconstruction, image registration and RGB color segmentation were automated using ImageJ software to avoid operator-dependent subjective errors. Based on the 3D tumor architecture, we classified the mode of invasion into four types: pushing and bulky architecture; trabecular architecture; diffuse spreading; and special forms. Direct visualization and quantitative assessment of the parenchymal-stromal border provide a new dimension in our understanding of OTSCC architecture. These 3D morphometric analyses also ascertained that cell invasion (individually and collectively) occurs at the deep invasive front of the OTSCC. These results demonstrate the advantages of histology-based 3D reconstruction for evaluating tumor architecture and its potential for a wide range of applications.

No MeSH data available.


Related in: MedlinePlus