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. 2023 Aug 18;13(1):124. doi: 10.1038/s41408-023-00899-3

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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 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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Fig. 7 — A Stained images from multiplex imaging mass cytometry (MxIMC) using Hyperion from a region of interest (ROI) from a representative FL specimen. The whole slide was stained with 26 metal-conjugated antibodies and ROI were selected for Hyperion to scan. The images were viewed and merged using the MCD viewer (Fluidigm). B Dot plots showing surface marker expression by CyTOF from the same patient in (A). C, D The UMAP and Heatmap plots of cells from ROI of a representative FL patient. The data was extracted from segmentation process and analyzed with the R program (Seurat). 11 clusters were generated and annotated based on phenotype. E A spatial point plot with each color representing and indicating the origin of clusters from (C, D). The point is the X & Y coordinate of the centroid of each cell. This plot was analyzed and generated with the Seurat package. F CD19 staining (in Green) (left) alongside a spatial point plot produced using SciMap and color matching (right). G Distance plots, log of micron boxplots, using all the CD19⁺/CD45RA⁺ cells as the basis of pairwise comparisons by distance. Each boxplot was the distribution of that population’s distance away from all the CD19⁺/CD45RA⁺ cells, within each sub-regional annotation.