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Subcellular Mass Spectrometry Imaging and Absolute Quantitative Analysis across Organelles

Journal article
Authors Aurélien Thomen
Neda Najafinobar
Florent Penen
Emma Kay
Pratik P. Upadhyay
Xianchan Li
Nhu TN Phan
Per Malmberg
Magnus Klarqvist
Shalini Andersson
Michael E. Kurczy
Andrew G Ewing
Published in ACS nano
Volume 14
Issue 4
Pages 4316-4325
Publication year 2020
Published at Department of Chemistry and Molecular Biology
National Center for Imaging Mass Spectrometry
Pages 4316-4325
Language en
Keywords mass spectrometry imaging, nanoscale, nanoSIMS, organelles, subcellular concentration
Subject categories Analytical Chemistry


Mass spectrometry imaging is a field that promises to become a mainstream bioanalysis technology by allowing the combination of single-cell imaging and subcellular quantitative analysis. The frontier of single-cell imaging has advanced to the point where it is now possible to compare the chemical contents of individual organelles in terms of raw or normalized ion signal. However, to realize the full potential of this technology, it is necessary to move beyond this concept of relative quantification. Here we present a nanoSIMS imaging method that directly measures the absolute concentration of an organelle-associated, isotopically labeled, pro-drug directly from a mass spectrometry image. This is validated with a recently developed nanoelectrochemistry method for single organelles. We establish a limit of detection based on the number of isotopic labels used and the volume of the organelle of interest, also offering this calculation as a web application. This approach allows subcellular quantification of drugs and metabolites, an overarching and previously unmet goal in cell science and pharmaceutical development.

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