A novel approach for monitoring the spatial distribution and quantitative analysis of micronutrients in plant tissues...

Quantitative imaging of plant tissues by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is hindered by the lack of matrix-matched calibration standards. Established approaches, such as gelatine or homogenised tissue blocks, do not replicate plant matrices accurately. Here, we introduce a nano-dispenser-based calibration strategy that deposits nanolitre volumes of elemental standards directly onto paraffin-embedded grain sections, exploiting the low endogenous metal content of the endosperm to generate in situ calibration curves. Calibration performance for Mg, Mn, Cu, Zn, and Mo was assessed using LA-ICP-MS imaging and Iolite 4 data processing. The method demonstrated excellent linearity (R² > 0.98), reproducibility across multiple grains, and sub-ppm limits of detection. Comparative analysis with in-house homogenised blocks and NIST wheat reference material confirmed superior accuracy and reproducibility of the nano-dispenser approach. As a proof-of-concept, we have applied the method for the quantitative imaging of metals in a whole barley grain section, and the results show excellent agreement with published data obtained by conventional liquid-mode ICP MS. The technique reported here provides a robust, scalable, solution for quantitative metallomics studies of plant tissues, enabling improved assessment of nutrient distribution and supporting the development of standardised protocols for LA-ICP-MS imaging.