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1. Self-supervised semantic segmentation of retinal pigment epithelium cells in flatmount fluorescent microscopy images

   https://doi.org/10.1093/bioinformatics/btad191  

   Yu, Hanyi ; Wang, Fusheng ; Teodoro, George ; Chen, Fan ; Guo, Xiaoyuan ; Nickerson, John M. ; Kong, Jun ; Peng, ed., Hanchuan ( April 2023 , Bioinformatics)

   Morphological analyses with flatmount fluorescent images are essential to retinal pigment epithelial (RPE) aging studies and thus require accurate RPE cell segmentation. Although rapid technology advances in deep learning semantic segmentation have achieved great success in many biomedical research, the performance of these supervised learning methods for RPE cell segmentation is still limited by inadequate training data with high-quality annotations.

   To address this problem, we develop a Self-Supervised Semantic Segmentation (S4) method that utilizes a self-supervised learning strategy to train a semantic segmentation network with an encoder–decoder architecture. We employ a reconstruction and a pairwise representation loss to make the encoder extract structural information, while we create a morphology loss to produce the segmentation map. In addition, we develop a novel image augmentation algorithm (AugCut) to produce multiple views for self-supervised learning and enhance the network training performance. To validate the efficacy of our method, we applied our developed S4 method for RPE cell segmentation to a large set of flatmount fluorescent microscopy images, we compare our developed method for RPE cell segmentation with other state-of-the-art deep learning approaches. Compared with other state-of-the-art deep learning approaches, our method demonstrates better performance in both qualitative and quantitative evaluations, suggesting its promising potential to support large-scale cell morphological analyses in RPE aging investigations.

   The codes and the documentation are available at: https://github.com/jkonglab/S4_RPE.

    

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2. A Morphological Post-Processing Approach for Overlapped Segmentation of Bacterial Cell Images

   https://doi.org/10.3390/make4040052  

   Abeyrathna, Dilanga ; Rauniyar, Shailabh ; Sani, Rajesh K ; Huang, Pei-Chi ( December 2022 , Machine Learning and Knowledge Extraction)

   Scanning electron microscopy (SEM) techniques have been extensively performed to image and study bacterial cells with high-resolution images. Bacterial image segmentation in SEM images is an essential task to distinguish an object of interest and its specific region. These segmentation results can then be used to retrieve quantitative measures (e.g., cell length, area, cell density) for the accurate decision-making process of obtaining cellular objects. However, the complexity of the bacterial segmentation task is a barrier, as the intensity and texture of foreground and background are similar, and also, most clustered bacterial cells in images are partially overlapping with each other. The traditional approaches for identifying cell regions in microscopy images are labor intensive and heavily dependent on the professional knowledge of researchers. To mitigate the aforementioned challenges, in this study, we tested a U-Net-based semantic segmentation architecture followed by a post-processing step of morphological over-segmentation resolution to achieve accurate cell segmentation of SEM-acquired images of bacterial cells grown in a rotary culture system. The approach showed an 89.52% Dice similarity score on bacterial cell segmentation with lower segmentation error rates, validated over several cell overlapping object segmentation approaches with significant performance improvement.

    

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3. Community-developed checklists for publishing images and image analyses

   https://doi.org/10.1038/s41592-023-01987-9  

   Schmied, Christopher ; Nelson, Michael S. ; Avilov, Sergiy ; Bakker, Gert-Jan ; Bertocchi, Cristina ; Bischof, Johanna ; Boehm, Ulrike ; Brocher, Jan ; Carvalho, Mariana T. ; Chiritescu, Catalin ; et al ( September 2023 , Nature Methods)

   Images document scientific discoveries and are prevalent in modern biomedical research. Microscopy imaging in particular is currently undergoing rapid technological advancements. However, for scientists wishing to publish obtained images and image-analysis results, there are currently no unified guidelines for best practices. Consequently, microscopy images and image data in publications may be unclear or difficult to interpret. Here, we present community-developed checklists for preparing light microscopy images and describing image analyses for publications. These checklists offer authors, readers and publishers key recommendations for image formatting and annotation, color selection, data availability and reporting image-analysis workflows. The goal of our guidelines is to increase the clarity and reproducibility of image figures and thereby to heighten the quality and explanatory power of microscopy data. 

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4. Deep learning provides high accuracy in automated chondrocyte viability assessment in articular cartilage using nonlinear optical microscopy

   https://doi.org/10.1364/BOE.417478  

   Chen, Xun ; Li, Yang ; Wyman, Nicole ; Zhang, Zheng ; Fan, Hongming ; Le, Michael ; Gannon, Steven ; Rose, Chelsea ; Zhang, Zhao ; Mercuri, Jeremy ; et al ( April 2021 , Biomedical Optics Express)

   Chondrocyte viability is a crucial factor in evaluating cartilage health. Most cell viability assays rely on dyes and are not applicable forin vivoor longitudinal studies. We previously demonstrated that two-photon excited autofluorescence and second harmonic generation microscopy provided high-resolution images of cells and collagen structure; those images allowed us to distinguish live from dead chondrocytes by visual assessment or by the normalized autofluorescence ratio. However, both methods require human involvement and have low throughputs. Methods for automated cell-based image processing can improve throughput. Conventional image processing algorithms do not perform well on autofluorescence images acquired by nonlinear microscopes due to low image contrast. In this study, we compared conventional, machine learning, and deep learning methods in chondrocyte segmentation and classification. We demonstrated that deep learning significantly improved the outcome of the chondrocyte segmentation and classification. With appropriate training, the deep learning method can achieve 90% accuracy in chondrocyte viability measurement. The significance of this work is that automated imaging analysis is possible and should not become a major hurdle for the use of nonlinear optical imaging methods in biological or clinical studies.

    

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5. Evaluating the relationship between magnetic resonance image quality metrics and deep learning‐based segmentation accuracy of brain tumors

   https://doi.org/10.1002/mp.17059  

   Muthusivarajan, Rajarajeswari ; Celaya, Adrian ; Yung, Joshua_P ; Long, James_P ; Viswanath, Satish_E ; Marcus, Daniel_S ; Chung, Caroline ; Fuentes, David ( April 2024 , Medical Physics)

   Magnetic resonance imaging (MRI) scans are known to suffer from a variety of acquisition artifacts as well as equipment‐based variations that impact image appearance and segmentation performance. It is still unclear whether a direct relationship exists between magnetic resonance (MR) image quality metrics (IQMs) (e.g., signal‐to‐noise, contrast‐to‐noise) and segmentation accuracy.

   Deep learning (DL) approaches have shown significant promise for automated segmentation of brain tumors on MRI but depend on the quality of input training images. We sought to evaluate the relationship between IQMs of input training images and DL‐based brain tumor segmentation accuracy toward developing more generalizable models for multi‐institutional data.

   We trained a 3D DenseNet model on the BraTS 2020 cohorts for segmentation of tumor subregions enhancing tumor (ET), peritumoral edematous, and necrotic and non‐ET on MRI; with performance quantified via a 5‐fold cross‐validated Dice coefficient. MRI scans were evaluated through the open‐source quality control tool MRQy, to yield 13 IQMs per scan. The Pearson correlation coefficient was computed between whole tumor (WT) dice values and IQM measures in the training cohorts to identify quality measures most correlated with segmentation performance. Each selected IQM was used to group MRI scans as “better” quality (BQ) or “worse” quality (WQ), via relative thresholding. Segmentation performance was re‐evaluated for the DenseNet model when (i) training on BQ MRI images with validation on WQ images, as well as (ii) training on WQ images, and validation on BQ images. Trends were further validated on independent test sets derived from the BraTS 2021 training cohorts.

   For this study, multimodal MRI scans from the BraTS 2020 training cohorts were used to train the segmentation model and validated on independent test sets derived from the BraTS 2021 cohort. Among the selected IQMs, models trained on BQ images based on inhomogeneity measurements (coefficient of variance, coefficient of joint variation, coefficient of variation of the foreground patch) and the models trained on WQ images based on noise measurement peak signal‐to‐noise ratio (SNR) yielded significantly improved tumor segmentation accuracy compared to their inverse models.

   Our results suggest that a significant correlation may exist between specific MR IQMs and DenseNet‐based brain tumor segmentation performance. The selection of MRI scans for model training based on IQMs may yield more accurate and generalizable models in unseen validation.

    

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