Dataset Information

Segmentation of Multiple Nuclei from Non-overlapping Immuno-histochemically Stained Histological Hepatic Images.

ABSTRACT: In this paper, we describe an algorithm for accurately segmenting multiple nudfclei from clumps of non-overlapping immuno-histochemically stained histological hepatic (liver) images. This problem is notoriously difficult because of the degree of presence of stains among the multi-nucleated cells, the poor contrast of cell cytoplasm, and the presence of mucus, blood, and inflammatory cells in the images. Hepatocellular carcinoma, characterized by cellular and nuclear enlargement, nuclear pleomorphism, and multi-nucleation, poses a prominent threat. Our proposed method addresses the aforementioned issues for an automated diagnosis system by judging the presence of multiple nuclei in a two-step process: the Quickhull algorithm defines the convex hull of each cell in the image and candidate nuclei regions are located with morphological operations. A combination of features containing local minima and shape-dependent features is extracted for the detection of single or multiple nuclei in each cell with a significant reduction in the number of false positives and false negatives providing an accuracy of 89.76%.

SUBMITTER: Kalinathan L

PROVIDER: S-EPMC9984616 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature

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Publications

Segmentation of Multiple Nuclei from Non-overlapping Immuno-histochemically Stained Histological Hepatic Images.

Kalinathan Lekshmi L Kathavarayan Ruba Soundar RS

Journal of digital imaging 20220802 1

In this paper, we describe an algorithm for accurately segmenting multiple nudfclei from clumps of non-overlapping immuno-histochemically stained histological hepatic (liver) images. This problem is notoriously difficult because of the degree of presence of stains among the multi-nucleated cells, the poor contrast of cell cytoplasm, and the presence of mucus, blood, and inflammatory cells in the images. Hepatocellular carcinoma, characterized by cellular and nuclear enlargement, nuclear pleomorp ...[more]

PMID: 35918474

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Project description:IntroductionAutomatic nuclear segmentation in digital microscopic tissue images can aid pathologists to extract high-quality features for nuclear morphometrics and other analyses. However, image segmentation is a challenging task in medical image processing and analysis. This study aimed to develop a deep learning-based method for nuclei segmentation of histological images for computational pathology.MethodsThe original U-Net model sometime has a caveat in exploring significant features. Herein, we present the Densely Convolutional Spatial Attention Network (DCSA-Net) model based on U-Net to perform the segmentation task. Furthermore, the developed model was tested on external multi-tissue dataset - MoNuSeg. To develop deep learning algorithms for well-segmenting nuclei, a large quantity of data are mandatory, which is expensive and less feasible. We collected hematoxylin and eosin-stained image data sets from two hospitals to train the model with a variety of nuclear appearances. Because of the limited number of annotated pathology images, we introduced a small publicly accessible data set of prostate cancer (PCa) with more than 16,000 labeled nuclei. Nevertheless, to construct our proposed model, we developed the DCSA module, an attention mechanism for capturing useful information from raw images. We also used several other artificial intelligence-based segmentation methods and tools to compare their results to our proposed technique.ResultsTo prioritize the performance of nuclei segmentation, we evaluated the model's outputs based on the Accuracy, Dice coefficient (DC), and Jaccard coefficient (JC) scores. The proposed technique outperformed the other methods and achieved superior nuclei segmentation with accuracy, DC, and JC of 96.4% (95% confidence interval [CI]: 96.2 - 96.6), 81.8 (95% CI: 80.8 - 83.0), and 69.3 (95% CI: 68.2 - 70.0), respectively, on the internal test data set.ConclusionOur proposed method demonstrates superior performance in segmenting cell nuclei of histological images from internal and external datasets, and outperforms many standard segmentation algorithms used for comparative analysis.

Project description:Study questionHow can we best achieve tissue segmentation and cell counting of multichannel-stained endometriosis sections to understand tissue composition?Summary answerA combination of a machine learning-based tissue analysis software for tissue segmentation and a deep learning-based algorithm for segmentation-independent cell identification shows strong performance on the automated histological analysis of endometriosis sections.What is known alreadyEndometriosis is characterized by the complex interplay of various cell types and exhibits great variation between patients and endometriosis subtypes.Study design, size, durationEndometriosis tissue samples of eight patients of different subtypes were obtained during surgery.Participants/materials, setting, methodsEndometriosis tissue was formalin-fixed and paraffin-embedded before sectioning and staining by (multiplex) immunohistochemistry. A 6-plex immunofluorescence panel in combination with a nuclear stain was established following a standardized protocol. This panel enabled the distinction of different tissue structures and dividing cells. Artificial intelligence-based tissue and cell phenotyping were employed to automatically segment the various tissue structures and extract quantitative features.Main results and the role of chanceAn endometriosis-specific multiplex panel comprised of PanCK, CD10, α-SMA, calretinin, CD45, Ki67, and DAPI enabled the distinction of tissue structures in endometriosis. Whereas a machine learning approach enabled a reliable segmentation of tissue substructure, for cell identification, the segmentation-free deep learning-based algorithm was superior.Limitations, reasons for cautionThe present analysis was conducted on a limited number of samples for method establishment. For further refinement, quantification of collagen-rich cell-free areas should be included which could further enhance the assessment of the extent of fibrotic changes. Moreover, the method should be applied to a larger number of samples to delineate subtype-specific differences.Wider implications of the findingsWe demonstrate the great potential of combining multiplex staining and cell phenotyping for endometriosis research. The optimization procedure of the multiplex panel was transferred from a cancer-related project, demonstrating the robustness of the procedure beyond the cancer context. This panel can be employed for larger batch analyses. Furthermore, we demonstrate that the deep learning-based approach is capable of performing cell phenotyping on tissue types that were not part of the training set underlining the potential of the method for heterogenous endometriosis samples.Study funding/competing interest(s)All funding was provided through departmental funds. The authors declare no competing interests.Trial registration numberN/A.

Dataset Information

Segmentation of Multiple Nuclei from Non-overlapping Immuno-histochemically Stained Histological Hepatic Images.

Publications

Segmentation of Multiple Nuclei from Non-overlapping Immuno-histochemically Stained Histological Hepatic Images.

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