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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

Three-dimensional view of individual cancer foci segmented at the invasion front. The size of the corresponding cancer mass is expressed in terms of diameter of a sphere having the same volume. The number in parentheses indicates the number of nuclei segmented from the cancer volume. Red: Ki67-positive nuclei; green: Ki67-negative nuclei (see text for nuclear segmentation).
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fig5: Three-dimensional view of individual cancer foci segmented at the invasion front. The size of the corresponding cancer mass is expressed in terms of diameter of a sphere having the same volume. The number in parentheses indicates the number of nuclei segmented from the cancer volume. Red: Ki67-positive nuclei; green: Ki67-negative nuclei (see text for nuclear segmentation).

Mentions: Table 3 shows the size distribution analysis of 966 discohesive cancer foci segmented at the invasive front of 14 OTSCC cases. In this analysis, the volume of each segmented cancer mass was determined in the reconstructed 3D space and, for the sake of simplicity, the size distribution was expressed in terms of the diameter of a sphere of the same volume. Notably, the majority of discohesive cancer foci were small in size, with the greatest population (39%) having a diameter ranging between 16 to 20 μm. To gain further insight into the cellular constitution of these discohesive cancer foci, we developed a computational algorithm using Ratoc TRI-SRF2 software to distinguish Ki67-positive and Ki67-negative nuclei in individual cancer foci. Ki67-negative nuclei were designated as open space circumscribed by CK-positive cytoplasm (see Figure 2(d)); noise images were deleted in the same way as applied for segmentation of Ki67-positive nuclei. To date, we have completed 3D analyses and visualization of 50 discohesive cancer foci randomly selected from different size ranges and OTSCC cases. Figure 5 shows representative 3D images of Ki67-positive (red) and Ki67-negative (green) nuclei in discohesive cancer foci. The smallest consisted of a single CK-positive cell with a Ki67-negative nucleus, while the largest analyzed so far included a total of 1,292 nuclei, comprising 277 Ki67-positive and 1,015 Ki67-negative nuclei. It is important to note that the majority (68%) of discohesive cancer foci with diameters <25 μm contained only a few cancer cells. In addition to highlighting the differences in the number of nuclei, 3D visualization of individual discohesive cancer foci disclosed their heterogeneous morphological features, for example, spheroidal, amoeboid, branching or stretching with extension of projections into the surrounding environment.


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)

Three-dimensional view of individual cancer foci segmented at the invasion front. The size of the corresponding cancer mass is expressed in terms of diameter of a sphere having the same volume. The number in parentheses indicates the number of nuclei segmented from the cancer volume. Red: Ki67-positive nuclei; green: Ki67-negative nuclei (see text for nuclear segmentation).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3818895&req=5

fig5: Three-dimensional view of individual cancer foci segmented at the invasion front. The size of the corresponding cancer mass is expressed in terms of diameter of a sphere having the same volume. The number in parentheses indicates the number of nuclei segmented from the cancer volume. Red: Ki67-positive nuclei; green: Ki67-negative nuclei (see text for nuclear segmentation).
Mentions: Table 3 shows the size distribution analysis of 966 discohesive cancer foci segmented at the invasive front of 14 OTSCC cases. In this analysis, the volume of each segmented cancer mass was determined in the reconstructed 3D space and, for the sake of simplicity, the size distribution was expressed in terms of the diameter of a sphere of the same volume. Notably, the majority of discohesive cancer foci were small in size, with the greatest population (39%) having a diameter ranging between 16 to 20 μm. To gain further insight into the cellular constitution of these discohesive cancer foci, we developed a computational algorithm using Ratoc TRI-SRF2 software to distinguish Ki67-positive and Ki67-negative nuclei in individual cancer foci. Ki67-negative nuclei were designated as open space circumscribed by CK-positive cytoplasm (see Figure 2(d)); noise images were deleted in the same way as applied for segmentation of Ki67-positive nuclei. To date, we have completed 3D analyses and visualization of 50 discohesive cancer foci randomly selected from different size ranges and OTSCC cases. Figure 5 shows representative 3D images of Ki67-positive (red) and Ki67-negative (green) nuclei in discohesive cancer foci. The smallest consisted of a single CK-positive cell with a Ki67-negative nucleus, while the largest analyzed so far included a total of 1,292 nuclei, comprising 277 Ki67-positive and 1,015 Ki67-negative nuclei. It is important to note that the majority (68%) of discohesive cancer foci with diameters <25 μm contained only a few cancer cells. In addition to highlighting the differences in the number of nuclei, 3D visualization of individual discohesive cancer foci disclosed their heterogeneous morphological features, for example, spheroidal, amoeboid, branching or stretching with extension of projections into the surrounding environment.

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