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. 2024 Jun 25;26:104. doi: 10.1186/s13058-024-01844-3

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© The Author(s) 2024

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — Autoradiography and immunofluorescence (IF) staining show that spatial heterogeneity and overall uptake are distinct measurements. A-B) Representative central slice of PET images showed high and low heterogeneity of [⁸⁹Zr]Zr-DFO-CD4 and [⁸⁹Zr]Zr-DFO-CD8 distribution, despite differences in total levels of uptake. C-D) Topographic map of [⁸⁹Zr]Zr-DFO-CD4 and [⁸⁹Zr]Zr-DFO-CD8 PET slices indicated regional hotspots. E-F) Corresponding autoradiography images immediately after PET imaging. G-H) Hematoxylin and eosin (H&E) staining of 4T1 tumor sections revealed that areas of high [⁸⁹Zr]Zr-DFO-CD4 or [⁸⁹Zr]Zr-DFO-CD8 uptake displayed a substantial concentration of densely packed tumor regions. I-J) IF staining for CD4 or CD8 in 4T1 tumor sections displayed a notable presence of CD4 + or CD8 + cells in proximity to high cellular tumor regions, a pattern consistent with the uptake pattern of the PET tracer