PreprintCited by 24Cell Image Analysis TechniquesSingle-cell and spatial transcriptomicsAI in cancer detection

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Automated classification of cellular expression in multiplexed imaging data with Nimbus

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Preprint — published as 10.1038/s41592-025-02826-9

Abstract

Multiplexed imaging offers a powerful approach to characterize the spatial topography of tissues in both health and disease. To analyze such data, the specific combination of markers that are present in each cell must be enumerated to enable accurate phenotyping, a process that often relies on unsupervised clustering. We constructed the Pan-Multiplex (Pan-M) dataset containing 197 million distinct annotations of marker expression across 15 different cell types. We used Pan-M to create Nimbus, a deep learning model to predict marker positivity from multiplexed image data. Nimbus is a pre-trained model that uses the underlying images to classify marker expression across distinct cell types, from different tissues, acquired using different microscope platforms, without requiring any retraining. We demonstrate that Nimbus predictions capture the underlying staining patterns of the full diversity of markers present in Pan-M. We then show how Nimbus predictions can be integrated with downstream clustering algorithms to robustly identify cell subtypes in image data. We have open-sourced Nimbus and Pan-M to enable community use at https://github.com/angelolab/Nimbus-Inference .

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Metadata

Authors
Rumberger, J. L., Greenwald, N. F., Ranek, J. S., Boonrat, P., Walker, C., Franzen, J., Varra, S. R., Kong, A., Sowers, C., Liu, C. C., Averbukh, I., Piyadasa, H., Vanguri, R., Nederlof, I., Wang, X. J., Van Valen, D., Kok, M., Hollmann, T. J., Kainmueller, D., & Angelo, M.
Affiliations
  • Max Delbrück Center (DE)
  • Stanford Medicine (US)
  • New York University (US)
  • The Netherlands Cancer Institute (NL)
  • California Institute of Technology (US)
  • Memorial Sloan Kettering Cancer Center (US)
  • University of Potsdam (DE)
Corresponding author
  • Greenwald, Noah F.
  • Angelo, Michael
Venue
Nature Methods
Publisher
openRxiv
Published
2024-06-03
Volume
22
Issue
10
Pages
2161-2170
ISSN
1548-7091 (ISSN-L)
OpenAlex
W4414930807
Cited by
24
References
1
Citations by year
2026: 15 · 2025: 7 · 2024: 2
FWCI
8.26
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Nature Methods (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Pure Amsterdam UMC (publishedVersion, other-oa)
  • MDC Repository (Max-Delbrueck-Center for Molecular Medicine) (acceptedVersion)
  • PubMed Central (submittedVersion, cc-by)
  • MDC Repository (Max-Delbrueck-Center for Molecular Medicine) (submittedVersion)
Funders
  • U.S. Department of Defense (W81XWH2110143)
  • U.S. Department of Health and Human Services (5U54CA20997105, CA264307)
  • Bill and Melinda Gates Foundation
  • Cancer Research Institute
  • Breast Cancer Research Foundation
  • Wellcome Trust
  • Deutscher Akademischer Austauschdienst
  • National Institutes of Health (5dp5od019822-05, 1r01ca240638-01a1, 5f99ca264307-02, 5r01ca229529-04, CA264307, DP5 OD019822, 4k00ca264307-03)
  • National Institute on Aging (R01AG056287, R01AG057915, R01AG068279)
  • National Heart, Lung, and Blood Institute (U54HL165445)
  • National Cancer Institute (CA264307, 5UH3CA24663303, 1R01CA24063801A1, 5U54CA20997105, 5R01CA22952904, UH3CA246633, U54CA209971, 1U24CA22430901, U24CA224309)
  • National Institute of Allergy and Infectious Diseases (F31AI165180, R01AI189963)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
  • Physical Sciences › Computer Science › Artificial Intelligence › AI in cancer detection
Keywords
multiplexed imaging, cell classification, cell phenotyping, cell subtypes, deep learning models, unsupervised clustering, spatial transcriptomics, pre-trained models, cell type annotation

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