Biology subjects

Kristensen, E. V.

Publications and source records attributed to Kristensen, E. V..

3 recordsLinked to original sources

bioRxiv2025

Achieving Micrometer-Scale 4D X-ray tomography of Living Leaf Tissue in the Laboratory

A methodology for achieving micrometer-scale 4D X-ray lab microscopy of living leaf tissue was developed to overcome challenges associated with delicate tissues, radiation damage, and motion artifacts during in vivo imaging. The study focused on optimizing laboratory based X-ray micro-computed tomography (microCT) parameters to balance high-resolution imaging with minimized physiological stress and radiation dose quantification. Assessing the dose-safe imaging window required comparing vertical and horizontal leaf mounting setups. Results demonstrated that the horizontal setup provided greater stability, preventing tissue degradation and maintaining sample viability during continuous acquisitions lasting up to 22 hours ([~]15600 Gy). MicroCT capacities were clearly able to resolve microstructures at the cellular level, achieving a pixel size down to 1 {micro}m. Furthermore, this optimized methodology confirmed the ability to track the spatiotemporal dynamics of applied compounds such as iohexol and aggregated nanoparticles within the leaf tissue. This work establishes that accessible laboratory based microCT enables the in vivo 4D monitoring of anatomical and physiological changes in living plants.

plant biology↗

bioRxiv2025

Foliar Application of Polymer-coated Manganese Dioxide Nanoparticles: Mechanisms of Uptake and Metabolic Responses in Manganese Deficient Barley

The application of nanotechnology in plant science is unlocking innovative approaches to enhance nutrient use efficiency in crops, particularly through foliar fertilization. This study demonstrates that colloidally stable, pH-responsive polyacrylic acid (PAA)-coated manganese dioxide (MnO2) nanoparticles (nPAA-MnO2) can be designed to significantly restore key metabolic functionalities in manganese (Mn)-deficient barley (Hordeum vulgare) within a few days. Using a combination of advanced bioimaging techniques - including confocal laser scanning microscopy (CLSM), laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS), and X-ray nano-computed tomography (nano-CT), we mapped the uptake and distribution pathways of nPAA-MnO2 compared to ionic Mn. While soluble Mn2+ ions primarily enter through hydrophilic cuticular pores, nPAA-MnO2 penetrates leaves via stomata, facilitated by the application of an organosilicone surfactant and glycerol to enhance wetting and hydraulic activation of stomatal pores. Within a few hours, nPAA-MnO2 accumulated in the sub-stomatal cavity and mesophyll apoplast, gradually releasing bioavailable Mn ions in the acidic apoplast environment. Moreover, labeling experiments with tracer ions revealed nPAA-MnO2 hot-spots around the vascular bundles and a limited but significant basipetal translocation of intact nanoparticles out of the foliar application zone, a pivotal step towards converting immobile nutrients such as Mn into mobile ones. Importantly, and unlike ionic Mn solutions, nPAA-MnO2 could be applied at high doses without causing leaf scorching and cytotoxicity, paving the way for more sustainable and efficient foliar fertilization practices. These novel aspects of nanoparticle uptake, translocation, and assimilation underscores the potential of nanotechnology to address nutrient mobility challenges in agriculture, representing an important contribution to the green transition of modern crop production.

plant biology↗

bioRxiv2025

3D X-ray microscopy lights up nanoparticles in plants

The discovery of novel plant fertilization strategies heavily relies on our capabilities to probe physiological processes in living plants with sub-cellular precision. State-of-the-art microscopy techniques are in general limited to surface investigation or they require elaborated tissue preparation and often destruction. X-ray microscopy has the potential to resolve some of these limitations by generating micro-to nanometer-scale 3D images deep into the tissue. We introduce experimental designs and the quantitative analysis methodologies, pioneering nanoscale ({approx}150 nm resolution) in-vivo 3D microscopy of plant tissue. We show the first direct in-vivo visualization of foliar-applied untagged nanoparticulate fertilizers deep under the leaf surface, not accessible by other microscopy methods. Ultimately, our approach provides the means for a direct observation of nanoparticle transport and dissolution in living plant tissue, a step critical for developing sustainable plant fertilization approaches.

plant biology↗