Plants produce a wide variety of metabolites that are heterogeneously distributed throughout the plant body. Accurate information on their spatial distribution, in addition to their overall abundance, is essential for understanding plant physiological processes and developmental regulation. Conventional mass spectrometric analyses provide precise quantification of metabolites but require tissue homogenization, resulting in the loss of spatial information. Mass spectrometry imaging (MSI) enables the direct, label-free visualization of molecules in plant tissue sections. By acquiring mass spectra in a spatially resolved manner, MSI allows the simultaneous detection and localization of multiple metabolites on a single tissue section, even in the absence of target-specific probes such as antibodies. Recent methodological advances have expanded the applicability of MSI in plant science, including the visualization of low-molecular-weight compounds such as plant hormones and secondary metabolite. In this chapter, we review recent progress in MSI-based approaches for spatial metabolite analysis in plants, with a particular focus on plant hormone imaging. We discuss the principles and applications of matrix-assisted laser desorption/ionization (MALDI) and nanoparticle-assisted laser desorption/ionization (Nano-PALDI) MSI, highlighting their advantages and limitations. Finally, we outline current technical challenges and future perspectives for the application of MSI in plant metabolomics and hormone research.