Mass Spectrometry Imaging: Unveiling New Horizons in Antibiotic Research

Mass spectrometry imaging (MSI) emerged as a revolutionary spatial molecular imaging approach for antibiotic spatial distribution and drug resistance mechanism, leveraging its capabilities for label-free, high-resolution and simultaneous multi-molecule detection. This study systematically reviews the progress of MSI technology, with a particular focus on the technological innovation of ionizations sources (MALDI, DESI, SIMS) and high-throughput intelligent data analysis systems; and explores the multi-dimensional applications in antibiotic research, highlighting its potential to reveal spatial and temporal dynamics of antibiotics through spatial pharmacokinetics (drug-metabolite colocalization, barrier penetration assessment, and optimization of nano-delivery systems). Furthermore, this study discusses the incorporation of MSI with tracking metabolic reprogramming within resistant bacteria, and mapping spatiotemporal drug-microbe interactions in resistant bacterial populations; and its expansion into microbial identification, environmental residue monitoring, and novel drug discovery and development. Despite the technical bottlenecks such as sensitivity, quantitative standardization and multimodal integration, MSI is anticipated to break through the paradigm of antibiotic research through the development of probes, the construction of clinical translational standards, and the fusion of multi-omics intelligence, providing precise solutions for the prevention and control of the drug resistance crisis. The aim of this paper is to provide a theoretical framework and technological insights for MSI-driven antibiotic research.