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CellSighter – A neural network to classify cells in highly multiplexed images

Yael Amitay, View ORCID ProfileYuval Bussi, Ben Feinstein, View ORCID ProfileShai Bagon, View ORCID ProfileIdan Milo, View ORCID ProfileLeeat Keren
doi: https://doi.org/10.1101/2022.11.07.515441
Yael Amitay
1Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel
2Department of Mathematics and Computer Science, Weizmann Institute of Science, Rehovot, Israel
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Yuval Bussi
1Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel
2Department of Mathematics and Computer Science, Weizmann Institute of Science, Rehovot, Israel
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Ben Feinstein
2Department of Mathematics and Computer Science, Weizmann Institute of Science, Rehovot, Israel
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Shai Bagon
2Department of Mathematics and Computer Science, Weizmann Institute of Science, Rehovot, Israel
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Idan Milo
1Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel
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Leeat Keren
1Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel
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  • For correspondence: c.m.a.van.ravenswaaij@umcg.nl
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Abstract

Multiplexed imaging enables measurement of multiple proteins in situ, offering an unprecedented opportunity to chart various cell types and states in tissues. However, cell classification, the task of identifying the type of individual cells, remains challenging, labor-intensive, and limiting to throughput. Here, we present CellSighter, a deep-learning based pipeline to accelerate cell classification in multiplexed images. Given a small training set of expert-labeled images, CellSighter outputs the label probabilities for all cells in new images. CellSighter achieves over 80% accuracy for major cell types across imaging platforms, which approaches inter-observer concordance. Ablation studies and simulations show that CellSighter is able to generalize its training data and learn features of protein expression levels, as well as spatial features such as subcellular expression patterns. CellSighter’s design reduces overfitting, and it can be trained with only thousands or even hundreds of labeled examples. CellSighter also outputs a prediction confidence, allowing downstream experts control over the results. Altogether, CellSighter drastically reduces hands-on time for cell classification in multiplexed images, while improving accuracy and consistency across datasets.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • https://github.com/KerenLab/CellSighter

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted November 08, 2022.
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CellSighter – A neural network to classify cells in highly multiplexed images
Yael Amitay, Yuval Bussi, Ben Feinstein, Shai Bagon, Idan Milo, Leeat Keren
bioRxiv 2022.11.07.515441; doi: https://doi.org/10.1101/2022.11.07.515441
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CellSighter – A neural network to classify cells in highly multiplexed images
Yael Amitay, Yuval Bussi, Ben Feinstein, Shai Bagon, Idan Milo, Leeat Keren
bioRxiv 2022.11.07.515441; doi: https://doi.org/10.1101/2022.11.07.515441

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