Addressing uncertainties in correlative imaging of exogenous particles with the tissue microanatomy with synchronous imaging strategies

Alexander P Morrell, Richard A Martin, Helen M Roberts, Hiram Castillo-Michel, J Frederick W Mosselmans, Kalotina Geraki, Adrian T Warfield, Paul Lingor, Wasif Qayyum, Daniel Graf, Maria Febbraio, Owen Addison*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Exposure to exogenous particles is of increasing concern to human health. Characterizing the concentrations, chemical species, distribution, and involvement of the stimulus with the tissue microanatomy is essential in understanding the associated biological response. However, no single imaging technique can interrogate all these features at once, which confounds and limits correlative analyses. Developments of synchronous imaging strategies, allowing multiple features to be identified simultaneously, are essential to assess spatial relationships between these key features with greater confidence. Here, we present data to first highlight complications of correlative analysis between the tissue microanatomy and elemental composition associated with imaging serial tissue sections. This is achieved by assessing both the cellular and elemental distributions in three-dimensional space using optical microscopy on serial sections and confocal X-ray fluorescence spectroscopy on bulk samples, respectively. We propose a new imaging strategy using lanthanide-Tagged antibodies with X-ray fluorescence spectroscopy. Using simulations, a series of lanthanide tags were identified as candidate labels for scenarios where tissue sections are imaged. The feasibility and value of the proposed approach are shown where an exposure of Ti was identified concurrently with CD45 positive cells at sub-cellular resolutions. Significant heterogeneity in the distribution of exogenous particles and cells can be present between immediately adjacent serial sections showing a clear need of synchronous imaging methods. The proposed approach enables elemental compositions to be correlated with the tissue microanatomy in a highly multiplexed and non-destructive manner at high spatial resolutions with the opportunity for subsequent guided analysis.

Original languageEnglish
Article numbermfad030
Number of pages9
Issue number6
Early online date16 May 2023
Publication statusPublished - Jun 2023

Bibliographical note

Copyright © The Author(s) 2023. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

Funding: The following funding is acknowledged: Natural Sciences and Engineering Research Council of Canada (grant No. 2017-05862).


  • Synchrotron X-ray fluorescence spectroscopy
  • correlative imaging
  • exogenous metal imaging
  • endogenous elemental imaging
  • metal labelled antibodies
  • lanthanide X-ray fluorescence
  • synchrotron confocal X-ray fluorescence spectroscopy


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