Untargeted pixel-by-pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imaging

Mass spectrometry imaging (MSI) is a powerful technology used to define the spatial distribution and relative abundance of metabolites across tissue cryosections. While software packages exist for pixel-by-pixel individual metabolite and limited target pairs of ratio imaging, the research community...

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Main Authors: Huiyong Cheng, Dawson Miller, Nneka Southwell, Paola Porcari, Joshua L Fischer, Isobel Taylor, J Michael Salbaum, Claudia Kappen, Fenghua Hu, Cha Yang, Kayvan R Keshari, Steven S Gross, Marilena D'Aurelio, Qiuying Chen
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Language:English
Published: eLife Sciences Publications Ltd 2025-03-01
Series:eLife
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Online Access:https://elifesciences.org/articles/96892
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author Huiyong Cheng
Dawson Miller
Nneka Southwell
Paola Porcari
Joshua L Fischer
Isobel Taylor
J Michael Salbaum
Claudia Kappen
Fenghua Hu
Cha Yang
Kayvan R Keshari
Steven S Gross
Marilena D'Aurelio
Qiuying Chen
author_facet Huiyong Cheng
Dawson Miller
Nneka Southwell
Paola Porcari
Joshua L Fischer
Isobel Taylor
J Michael Salbaum
Claudia Kappen
Fenghua Hu
Cha Yang
Kayvan R Keshari
Steven S Gross
Marilena D'Aurelio
Qiuying Chen
author_sort Huiyong Cheng
collection DOAJ
description Mass spectrometry imaging (MSI) is a powerful technology used to define the spatial distribution and relative abundance of metabolites across tissue cryosections. While software packages exist for pixel-by-pixel individual metabolite and limited target pairs of ratio imaging, the research community lacks an easy computing and application tool that images any metabolite abundance ratio pairs. Importantly, recognition of correlated metabolite pairs may contribute to the discovery of unanticipated molecules in shared metabolic pathways. Here, we describe the development and implementation of an untargeted R package workflow for pixel-by-pixel ratio imaging of all metabolites detected in an MSI experiment. Considering untargeted MSI studies of murine brain and embryogenesis, we demonstrate that ratio imaging minimizes systematic data variation introduced by sample handling, markedly enhances spatial image contrast, and reveals previously unrecognized metabotype-distinct tissue regions. Furthermore, ratio imaging facilitates identification of novel regional biomarkers and provides anatomical information regarding spatial distribution of metabolite-linked biochemical pathways. The algorithm described herein is applicable to any MSI dataset containing spatial information for metabolites, peptides or proteins, offering a potent hypothesis generation tool to enhance knowledge obtained from current spatial metabolite profiling technologies.
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spelling doaj-art-cf9c6749dd1b455391cf948fb5bb82cd2025-08-20T03:01:50ZengeLife Sciences Publications LtdeLife2050-084X2025-03-011310.7554/eLife.96892Untargeted pixel-by-pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imagingHuiyong Cheng0Dawson Miller1Nneka Southwell2Paola Porcari3Joshua L Fischer4Isobel Taylor5J Michael Salbaum6Claudia Kappen7Fenghua Hu8https://orcid.org/0000-0002-6447-9992Cha Yang9Kayvan R Keshari10Steven S Gross11Marilena D'Aurelio12Qiuying Chen13https://orcid.org/0000-0001-5909-3959Department of Pharmacology, Weill Cornell Medicine, New York, United StatesDepartment of Pharmacology, Weill Cornell Medicine, New York, United StatesBrain and Mind Research Institute, Weill Cornell Medicine, New York City, United StatesMemorial Sloan Kettering Cancer Center, New York, United StatesBruker Daltonics, Billerica, United StatesDepartment of Pharmacology, Weill Cornell Medicine, New York, United StatesPennington Biomedical Research Center, Louisiana State University, Baton Rouge, United StatesPennington Biomedical Research Center, Louisiana State University, Baton Rouge, United StatesCornell University, Department of Molecular Biology & Genetics, Ithaca, United StatesCornell University, Department of Molecular Biology & Genetics, Ithaca, United StatesMemorial Sloan Kettering Cancer Center, New York, United StatesDepartment of Pharmacology, Weill Cornell Medicine, New York, United StatesBrain and Mind Research Institute, Weill Cornell Medicine, New York City, United StatesDepartment of Pharmacology, Weill Cornell Medicine, New York, United StatesMass spectrometry imaging (MSI) is a powerful technology used to define the spatial distribution and relative abundance of metabolites across tissue cryosections. While software packages exist for pixel-by-pixel individual metabolite and limited target pairs of ratio imaging, the research community lacks an easy computing and application tool that images any metabolite abundance ratio pairs. Importantly, recognition of correlated metabolite pairs may contribute to the discovery of unanticipated molecules in shared metabolic pathways. Here, we describe the development and implementation of an untargeted R package workflow for pixel-by-pixel ratio imaging of all metabolites detected in an MSI experiment. Considering untargeted MSI studies of murine brain and embryogenesis, we demonstrate that ratio imaging minimizes systematic data variation introduced by sample handling, markedly enhances spatial image contrast, and reveals previously unrecognized metabotype-distinct tissue regions. Furthermore, ratio imaging facilitates identification of novel regional biomarkers and provides anatomical information regarding spatial distribution of metabolite-linked biochemical pathways. The algorithm described herein is applicable to any MSI dataset containing spatial information for metabolites, peptides or proteins, offering a potent hypothesis generation tool to enhance knowledge obtained from current spatial metabolite profiling technologies.https://elifesciences.org/articles/96892brainembryoadipose
spellingShingle Huiyong Cheng
Dawson Miller
Nneka Southwell
Paola Porcari
Joshua L Fischer
Isobel Taylor
J Michael Salbaum
Claudia Kappen
Fenghua Hu
Cha Yang
Kayvan R Keshari
Steven S Gross
Marilena D'Aurelio
Qiuying Chen
Untargeted pixel-by-pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imaging
eLife
brain
embryo
adipose
title Untargeted pixel-by-pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imaging
title_full Untargeted pixel-by-pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imaging
title_fullStr Untargeted pixel-by-pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imaging
title_full_unstemmed Untargeted pixel-by-pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imaging
title_short Untargeted pixel-by-pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imaging
title_sort untargeted pixel by pixel metabolite ratio imaging as a novel tool for biomedical discovery in mass spectrometry imaging
topic brain
embryo
adipose
url https://elifesciences.org/articles/96892
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