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. 2017 Apr 4;7:603. doi: 10.1038/s41598-017-00703-w

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Insight into the complexity of the high mass resolution MSI data: (a) Comparison of the average mass spectra obtained from consecutive coronal mouse brain tissue sections with MALDI–ToF MSI and MALDI–FTICR MSI. (b) Scanned image of the Nissl-stained mouse brain tissue, the solid black line highlights the tumor. (c) From left to right: average MALDI TOF mass spectrum (left), average MALDI FTICR mass spectrum (center), and overlay of the average MALDI FTICR mass spectra from the tumor (pink) and healthy (blue) regions. The shaded region highlights tumor specific proteoforms that could only be revealed with the MALDI-FTICR. (d) The ultra-high resolution of the MALDI-FTICR instrument distinguished two doubly charged proteoforms of Histone H2B and a different singly charged protein. (e) MALDI-FTICR spectra showing the differentiation of two protein ions of similar mass with interspersed and isobaric isotopomers, and which have differential localization. (f) Overlaid average mass spectra of tumor and healthy regions showing the specific distribution of three ions that could not be detected with MALDI-TOF MSI (MALDI-TOF mass spectrum shown in insert).