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Correlative Imaging of Structural Biochemistry in Plant and Food Quality Research Within an Interoperable Data Acquisition Platform

Marjana Regvar · Boštjan Jenčič · Martin Šala · Aleš Kladnik · Iztok Dogša · Maja Koblar · Mitja Kelemen · Primož Vavpetič · Katarina Vogel-Mikuš · Ivan Kreft · Primož Pelicon · Paula Pongrac

Microscopy and Microanalysis · 15 Jul 2025 · 10.1093/mam/ozaf057

Abstract

Abstract Correlative imaging is a powerful tool for revealing information on cell-type structures and their biochemistry, with the potential to inform healthier food choices and improved dietary recommendations. Determination of plant structures and their structural biochemistry advances our understanding of specific structures designed to store different biomolecules within cells and tissues. Compared to the classical biochemical separation techniques, the key advantage of sequential correlative imaging techniques is in relating spatial plant (micro)structures to their biochemistry in a nondestructive manner. Sequential imaging reported here comprises six methodologies on a single sample, a cross-section of a Tartary buckwheat (Fagopyrum tataricum) grain, namely, bright-field and autofluorescence microscopy, fluorescence microspectroscopy, MeV-secondary ion mass spectrometry, micro-particle-induced X-ray emission, scanning electron microscopy coupled with energy dispersive X-ray spectroscopy, and laser ablation-inductively coupled plasma-mass spectrometry. Results confirm that the stepwise addition of the desired information across several classes of biomolecules and several spatial scales informs the quality and safety of plant-based produce across scales. Therefore, a viable workflow is proposed, enabling sequential spatial analysis of grain and highlighting plant structures' in situ specificity. The advantages and disadvantages of the selected methodologies were critically evaluated.

Code and data availability

The paper explicitly points to a public Zenodo deposit containing the correlative imaging data (SEM, micro-PIXE, MeV-SIMS maps) used in its analyses, with instructions for reproducing image fusion in Wolfram Mathematica or ImageJ.

Datasetpublic

sed to reveal the allocation of K to cotyledons (Supplementary Fused Image 1). Similarly, on the same SEM image, MeV-SIMS distribution maps under the selected peak were overlaid (Supplementary Fused Image 2). Custom combinations can be done in the Wolfram Mathematica program or in ImageJ (Merge Channels) using data available at https://doi.org/10.5281/zenodo.14628251, fol­ lowing the instructions in the Materials and Methods. Conclusions The low emission properties of fluorescence biomolecules, when excited with 405 nm light, inherently limit the informa­ tion acquired using fluorescence imaging. At this excitation wavelength, catechin may be the primary fluorophore in Tartary buckwheat cot

Open resource ↗zenodo · 10.5281/zenodo.14628251 · pdf-raw-page:13 lines:1-89