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. 2023 Oct 16;415(28):7011–7024. doi: 10.1007/s00216-023-04983-2

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Combination of MALDI-MSI and IMC on human breast cancer tissue. A Whole tissue H&E and MALDI-MSI images of breast samples processed by MALDI-MSI first (left column) and IMC first (right column) both taken with 80-µm step size. Images from two N-glycan structures (1809.625 and 2174.763 m/z) are shown followed by two extracellular matrix molecules (1034.572 m/z and 1125.529 m/z). B Magnified H&E and higher resolution (step size of 20 µm) MALDI-MSI images of area analyzed by IMC that shows the spot of tissue ablation from the IMC laser for matched N-glycan/peptide structures from A. C IMC images from slides processed by IMC first (left column) and MALDI-MSI first (right column). Markers visualized are labeled by row. D Comparison of N-glycan spectra showing MALDI-MSI first on top and IMC first flipped on the bottom. E Differences in peak intensity relative to MALDI-MSI first. F Top 10 peaks of highest intensity in IMC first are compared for N-glycans. G Comparisons of the whole spectra collagens. H The percent difference in peak intensity for a subset of selected peaks with respect to MALDI-MSI first. I Ten highest intensity peaks ranked by IMC first compared to intensities of MALDI-MSI first. (D–I are measured from the whole tissue)