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      SCIMAP: A Python Toolkit for Integrated Spatial Analysis of Multiplexed Imaging Data

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      Journal of open source software

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          Summary

          Multiplexed imaging data are revolutionizing our understanding of the composition and organization of tissues and tumors ( “Catching up with Multiplexed Tissue Imaging,” 2022). A critical aspect of such “tissue profiling” is quantifying the spatial relationships among cells at different scales from the interaction of neighboring cells to recurrent communities of cells of multiple types. This often involves statistical analysis of 10 7 or more cells in which up to 100 biomolecules (commonly proteins) have been measured. While software tools currently cater to the analysis of spatial transcriptomics data ( Liu et al., 2022), there remains a need for toolkits explicitly tailored to the complexities of multiplexed imaging data including the need to seamlessly integrate image visualization with data analysis and exploration. We introduce SCIMAP, a Python package specifically crafted to address these challenges. With SCIMAP, users can efficiently preprocess, analyze, and visualize large datasets, facilitating the exploration of spatial relationships and their statistical significance. SCIMAP’s modular design enables the integration of new algorithms, enhancing its capabilities for spatial analysis.

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          Cellpose: a generalist algorithm for cellular segmentation

          Many biological applications require the segmentation of cell bodies, membranes and nuclei from microscopy images. Deep learning has enabled great progress on this problem, but current methods are specialized for images that have large training datasets. Here we introduce a generalist, deep learning-based segmentation method called Cellpose, which can precisely segment cells from a wide range of image types and does not require model retraining or parameter adjustments. Cellpose was trained on a new dataset of highly varied images of cells, containing over 70,000 segmented objects. We also demonstrate a three-dimensional (3D) extension of Cellpose that reuses the two-dimensional (2D) model and does not require 3D-labeled data. To support community contributions to the training data, we developed software for manual labeling and for curation of the automated results. Periodically retraining the model on the community-contributed data will ensure that Cellpose improves constantly.
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            Deep Profiling of Mouse Splenic Architecture with CODEX Multiplexed Imaging

            Summary A highly multiplexed cytometric imaging approach, termed co-detection by indexing (CODEX), is used here to create multiplexed datasets of normal and lupus (MRL/lpr) murine spleens. CODEX iteratively visualizes antibody binding events using DNA barcodes, fluorescent dNTP analogs, and an in situ polymerization-based indexing procedure. An algorithmic pipeline for single-cell antigen quantification in tightly packed tissues was developed and used to overlay well-known morphological features with de novo characterization of lymphoid tissue architecture at a single-cell and cellular neighborhood levels. We observed an unexpected, profound impact of the cellular neighborhood on the expression of protein receptors on immune cells. By comparing normal murine spleen to spleens from animals with systemic autoimmune disease (MRL/lpr), extensive and previously uncharacterized splenic cell-interaction dynamics in the healthy versus diseased state was observed. The fidelity of multiplexed spatial cytometry demonstrated here allows for quantitative systemic characterization of tissue architecture in normal and clinically aberrant samples.
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              Highly multiplexed imaging of tumor tissues with subcellular resolution by mass cytometry.

              Mass cytometry enables high-dimensional, single-cell analysis of cell type and state. In mass cytometry, rare earth metals are used as reporters on antibodies. Analysis of metal abundances using the mass cytometer allows determination of marker expression in individual cells. Mass cytometry has previously been applied only to cell suspensions. To gain spatial information, we have coupled immunohistochemical and immunocytochemical methods with high-resolution laser ablation to CyTOF mass cytometry. This approach enables the simultaneous imaging of 32 proteins and protein modifications at subcellular resolution; with the availability of additional isotopes, measurement of over 100 markers will be possible. We applied imaging mass cytometry to human breast cancer samples, allowing delineation of cell subpopulations and cell-cell interactions and highlighting tumor heterogeneity. Imaging mass cytometry complements existing imaging approaches. It will enable basic studies of tissue heterogeneity and function and support the transition of medicine toward individualized molecularly targeted diagnosis and therapies.
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                Author and article information

                Journal
                101708638
                46693
                J Open Source Softw
                J Open Source Softw
                Journal of open source software
                2475-9066
                31 May 2024
                2024
                29 May 2024
                13 June 2024
                : 9
                : 97
                : 6604
                Affiliations
                [1 ]Department of Dermatology, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, United States of America
                [2 ]Ludwig Center for Cancer Research at Harvard, Harvard Medical School, Boston, MA, United States of America
                [3 ]Laboratory of Systems Pharmacology, Harvard Medical School, Boston, MA, United States of America
                Author notes
                []Corresponding author
                Author information
                http://orcid.org/0000-0003-4139-0618
                http://orcid.org/0000-0002-3364-1838
                Article
                NIHMS1998254
                10.21105/joss.06604
                11173324
                38873023
                4ba54571-1989-41db-a8b3-9f5e693f7a2e

                License: Authors of papers retain copyright and release the work under a Creative Commons Attribution 4.0 International License ( CC BY 4.0).

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