Abstract
AbstractMatrix‐assisted laser/desorption ionisation‐mass spectrometry imaging (MALDI‐MSI) enables label‐free imaging of biomolecules in biological tissues. However, many molecules remain undetected due to their poor ionisation efficiencies. These poor ionisation efficiencies practically limit spatial resolution. Herein, we address this challenge for aromatic antioxidants by reporting an innovative approach involving sequential matrix‐assisted laser desorption and two‐photon ionisation of desorbed neutrals. It is shown that ion yields increase with reduced sampling areas obtained using sub‐threshold primarily laser fluence. This counterintuitive observation could arise from a reduction in radical/ion neutralisation reactions within the sparse plume and/or favorable molecular desorption under low fluence conditions. The utility of this approach is demonstrated for imaging tocopherols and ubiquinols in mouse brain and prostate cancer tissue. This can pave the way for improved sensitivity in MSI experiments at cellular and sub‐cellular resolutions.
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