Integration of mass cytometry and single-cell RNA-sequencing of cells in bronchoalveolar lavage

Single-cell RNA sequencing (sc-RNA-seq) is a popular method for characterization of cell populations. However, the relationship between RNA and protein expression in cells is often discordant. Protein-based detection methods, such as cytometry by time-of-flight (CyTOF), can provide complementary dat...

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Main Authors: Elizabeth Guinto, Sameeksha Chopra, I-Chih Kuo, Samuel B. Shin, Melina Messing, Chung Y. Cheung, Julia SW. Yang, Firoozeh V. Gerayeli, William Yip, Stephen Milne, Rachel L. Eddy, Janice M. Leung, Kelly M. McNagny, Don D. Sin
Format: Article
Language:English
Published: Taylor & Francis Group 2025-04-01
Series:BioTechniques
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Online Access:https://www.tandfonline.com/doi/10.1080/07366205.2025.2505347
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author Elizabeth Guinto
Sameeksha Chopra
I-Chih Kuo
Samuel B. Shin
Melina Messing
Chung Y. Cheung
Julia SW. Yang
Firoozeh V. Gerayeli
William Yip
Stephen Milne
Rachel L. Eddy
Janice M. Leung
Kelly M. McNagny
Don D. Sin
author_facet Elizabeth Guinto
Sameeksha Chopra
I-Chih Kuo
Samuel B. Shin
Melina Messing
Chung Y. Cheung
Julia SW. Yang
Firoozeh V. Gerayeli
William Yip
Stephen Milne
Rachel L. Eddy
Janice M. Leung
Kelly M. McNagny
Don D. Sin
author_sort Elizabeth Guinto
collection DOAJ
description Single-cell RNA sequencing (sc-RNA-seq) is a popular method for characterization of cell populations. However, the relationship between RNA and protein expression in cells is often discordant. Protein-based detection methods, such as cytometry by time-of-flight (CyTOF), can provide complementary data to sc-RNA-seq. We collected bronchoalveolar lavage (BAL) from healthy participants and co-evaluated cell populations and gene/protein expression by applying sc-RNA-seq and CyTOF to the same samples. Cell populations were well correlated between these two platforms, but differences emerged at the sub-population level. Notably, macrophage subtypes did not correlate well; whereas T-lymphocytes did. Gene and protein expression levels were significantly correlated (p < .01). Overall, we recommend CyTOF as a tool to validate sc-RNA-seq data for select proteins and cell populations in BAL samples.
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institution Kabale University
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1940-9818
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publishDate 2025-04-01
publisher Taylor & Francis Group
record_format Article
series BioTechniques
spelling doaj-art-1f5238c9ba8847d3969e75fb2ff29c482025-08-20T03:44:17ZengTaylor & Francis GroupBioTechniques0736-62051940-98182025-04-0177417919010.1080/07366205.2025.2505347Integration of mass cytometry and single-cell RNA-sequencing of cells in bronchoalveolar lavageElizabeth Guinto0Sameeksha Chopra1I-Chih Kuo2Samuel B. Shin3Melina Messing4Chung Y. Cheung5Julia SW. Yang6Firoozeh V. Gerayeli7William Yip8Stephen Milne9Rachel L. Eddy10Janice M. Leung11Kelly M. McNagny12Don D. Sin13Centre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaSchool of Biomedical Engineering, University of British Columbia, Vancouver, CanadaSchool of Biomedical Engineering, University of British Columbia, Vancouver, CanadaSchool of Biomedical Engineering, University of British Columbia, Vancouver, CanadaSchool of Biomedical Engineering, University of British Columbia, Vancouver, CanadaCentre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaCentre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaCentre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaCentre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaSydney Medical School, The University of Sydney, Sydney, AustraliaCentre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaCentre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaCentre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaCentre for Heart Lung Innovation, St. Paul’s Hospital, University of British Columbia, Vancouver, CanadaSingle-cell RNA sequencing (sc-RNA-seq) is a popular method for characterization of cell populations. However, the relationship between RNA and protein expression in cells is often discordant. Protein-based detection methods, such as cytometry by time-of-flight (CyTOF), can provide complementary data to sc-RNA-seq. We collected bronchoalveolar lavage (BAL) from healthy participants and co-evaluated cell populations and gene/protein expression by applying sc-RNA-seq and CyTOF to the same samples. Cell populations were well correlated between these two platforms, but differences emerged at the sub-population level. Notably, macrophage subtypes did not correlate well; whereas T-lymphocytes did. Gene and protein expression levels were significantly correlated (p < .01). Overall, we recommend CyTOF as a tool to validate sc-RNA-seq data for select proteins and cell populations in BAL samples.https://www.tandfonline.com/doi/10.1080/07366205.2025.2505347Bronchoalveolar lavagemass cytometryproteinrNAsingle-cell RNA sequencing
spellingShingle Elizabeth Guinto
Sameeksha Chopra
I-Chih Kuo
Samuel B. Shin
Melina Messing
Chung Y. Cheung
Julia SW. Yang
Firoozeh V. Gerayeli
William Yip
Stephen Milne
Rachel L. Eddy
Janice M. Leung
Kelly M. McNagny
Don D. Sin
Integration of mass cytometry and single-cell RNA-sequencing of cells in bronchoalveolar lavage
BioTechniques
Bronchoalveolar lavage
mass cytometry
protein
rNA
single-cell RNA sequencing
title Integration of mass cytometry and single-cell RNA-sequencing of cells in bronchoalveolar lavage
title_full Integration of mass cytometry and single-cell RNA-sequencing of cells in bronchoalveolar lavage
title_fullStr Integration of mass cytometry and single-cell RNA-sequencing of cells in bronchoalveolar lavage
title_full_unstemmed Integration of mass cytometry and single-cell RNA-sequencing of cells in bronchoalveolar lavage
title_short Integration of mass cytometry and single-cell RNA-sequencing of cells in bronchoalveolar lavage
title_sort integration of mass cytometry and single cell rna sequencing of cells in bronchoalveolar lavage
topic Bronchoalveolar lavage
mass cytometry
protein
rNA
single-cell RNA sequencing
url https://www.tandfonline.com/doi/10.1080/07366205.2025.2505347
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