Comparison of Mid‐Infrared and Ultraviolet Lasers Coupled to the MALDESI Source for the Detection of Secondary Metabolites and Structural Lipids in <scp><i>Arabidopsis thaliana</i></scp>

Matrix-assisted laser desorption electrospray ionization (MALDESI) conventionally utilizes a mid-infrared (IR) laser for the desorption of neutrals, allowing for detection of hundreds to thousands of analytes simultaneously. This platform enables mass spectrometry imaging (MSI) capabilities to not only detect specific molecules but also reveal the distribution and localization of a wide range of biomolecules across an organism. However, an IR laser comes with its disadvantages when imaging plants. At a mid-IR wavelength (2970 nm), the compartmentalized endogenous water within the leaf structure acts as an internal matrix, causing rapid heating, and, in turn, degrades the spatial resolution and signal quality. An ultraviolet (UV) laser operates at wavelengths that overlap with the absorption bands of secondary metabolites allowing them to serve as sacrificial matrix molecules. With the integration and optimization of a 355 nm UV laser into the MALDESI-MSI NextGen source for the analysis of plants, we were able to detect diverse molecular classes including flavonoids, fatty acid derivatives, galactolipids, and glucosinolates, at higher ion abundances when compared to the mid-IR laser. These results show that re-visiting UV-MALDESI-MSI, without the need for an exogenous matrix, provides a promising approach for the detection and imaging of important analytes in plants.