Multimodal and multiscale correlative elemental imaging: From whole tissues down to organelles

Roudeau, Stéphane; Carmona, Asunción; Ortega, Richard

doi:10.1016/j.cbpa.2023.102372

Cited by 4 publications

(2 citation statements)

References 44 publications

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“…However, to obtain subcellular information, analytical techniques with higher spatial resolution need to be implemented. Multimodal correlative approaches are being developed for this purpose, often based on synchrotron X-ray fluorescence (SXRF) for metal nanoimaging. , SXRF spectrometry is a very powerful analytical tool offering high detection sensitivity, high spatial resolution on dedicated beamlines (<100 nm), and quantitative capability for metal mapping in biological samples . Although SXRF cannot be performed on living cells due to cellular damage caused by intense irradiation, SXRF can be performed under cryogenic conditions, enabling imaging of the native intracellular distribution of elements. − …”

Section: Introductionmentioning

confidence: 99%

“…SXRF imaging can be combined with other microscopy or spectro-imaging techniques to complement the information acquired on metal distribution with biological indicators such as cell compartments and organelles. ,,,− More specifically, knowing the colocalization of metals with proteins at the subcellular level is an important piece of information for understanding metal–protein interactions in cells. ,− However, the protocols usually employed in cell biology for imaging proteins with fluorescently labeled antibodies are based on chemical fixation and are known to alter the quantitative content and distribution of the elements. ,,− To avoid this denaturing step of chemical fixation, protein labeling can be performed with fluorescent dyes developed for live-cell microcopy. We have designed correlative approaches for imaging proteins by fluorescence light microscopy (FLM) in living cells and subsequent imaging of chemical elements by SXRF. ,,,, Live-cell FLM can be performed using epifluorescence, confocal or super-resolution microscopy, characterized by different spatial resolutions ranging from submicron to tenth-nanometer scale.…”

Section: Introductionmentioning

confidence: 99%

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Native Cryo-Correlative Light and Synchrotron X-ray Fluorescence Imaging of Proteins and Essential Metals in Subcellular Neuronal Compartments

Ortega,

Fernández-Monreal,

Pied

et al. 2024

Chemical & Biomedical Imaging

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Essential metals such as iron, copper, and zinc are required for a wide variety of biological processes. For example, they act as cofactors in many proteins, conferring enzymatic activity or structural stability. Interactions between metals and proteins are often difficult to characterize due to the low concentration of metals in biological tissues and the sometimes labile nature of the chemical bonds involved. To better understand the cellular functions of essential metals, we correlate protein localization, using fluorescence light microscopy (FLM), and metal distribution with synchrotron X-ray fluorescence (SXRF), a highsensitivity and high-spatial-resolution technique for metal imaging. Both chemical imaging modalities are implemented under cryogenic conditions to preserve native cell structure and chemical element distribution. As a proof of concept, we applied cryo-FLM and cryo-SXRF correlative imaging to cultured primary hippocampal neurons. Neurons were labeled under live conditions with fluorescent F-actin and tubulin dyes, then samples were flash-frozen and observed in a frozen hydrated state. This methodology, cryo-FLM combined to cryo-SXRF, revealed the distribution of iron, copper and zinc relative to F-actin and tubulin in the growth cones, dendrites, axons, and axonal en passant boutons of developing neurons.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Native Cryo-Correlative Light and Synchrotron X-ray Fluorescence Imaging of Proteins and Essential Metals in Subcellular Neuronal Compartments

Ortega,

Fernández-Monreal,

Pied

et al. 2024

Chemical & Biomedical Imaging

Self Cite

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Atomic spectrometry update: review of advances in X-ray fluorescence spectrometry and its special applications

Vanhoof,

Bacon,

Fittschen

et al. 2024

J. Anal. At. Spectrom.

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Integration of Elemental Imaging and Spatial Transcriptomic Profiling for Proof-of-Concept Metals-Based Pathway Analysis of Colon Tumor Microenvironment

Srivastava,

Shaik,

et al. 2024

Preprint

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The complex interplay between metal abundance, transport mechanisms, cell distribution, and tumor progression-related biological pathways (e.g., metabolism, collagen remodeling) remains poorly understood. Traditionally, genes and metals have been studied in isolation, limiting insights into their interactions. Recent advances in spatial transcriptomics and elemental profiling now enable comprehensive exploration of tissue-wide metal-gene interactions, though integration remains challenging. In this proof-of-concept study, we investigated metal-dependent signaling within the tumor microenvironment of a unique colorectal cancer (CRC) tumor. We implemented a spatial multimodal workflow which integrated elemental imaging, gene expression, cellular composition, and histopathological features to uncover metals-related pathways through spatially resolved differential expression analysis. Preliminary findings revealed significant associations, for instance: elevated iron correlated with mesenchymal phenotypes located at the tumor's proliferative front, reflecting epithelial-to-mesenchymal transition pathways, and extracellular matrix remodeling. High concentrations of copper were predominantly localized in regions of active tumor growth and associated with upregulation of immune response genes. This proof-of-concept workflow demonstrates the feasibility of integrating elemental imaging with spatial transcriptomics to identify metals-based gene correlates. Future application of this workflow to larger patient cohorts will pave the way for expansive comparisons across the metallome and transcriptome, ultimately identifying novel targets for tumor progression biomarkers and therapeutic interventions.

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Multimodal and multiscale correlative elemental imaging: From whole tissues down to organelles

Cited by 4 publications

References 44 publications

Native Cryo-Correlative Light and Synchrotron X-ray Fluorescence Imaging of Proteins and Essential Metals in Subcellular Neuronal Compartments

Native Cryo-Correlative Light and Synchrotron X-ray Fluorescence Imaging of Proteins and Essential Metals in Subcellular Neuronal Compartments

Atomic spectrometry update: review of advances in X-ray fluorescence spectrometry and its special applications

Integration of Elemental Imaging and Spatial Transcriptomic Profiling for Proof-of-Concept Metals-Based Pathway Analysis of Colon Tumor Microenvironment

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