Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) is an important technique for the spatially resolved molecular analysis of tissue sections. The selection of matrices influences the resulting mass spectra to a high degree. For extensive and simultaneous analysis, the application of different matrices to one tissue sample is desirable. To date, only a single matrix could be applied to a tissue section per experiment. However, repetitive removal of the matrix makes this approach time-consuming and damaging to tissue samples. To overcome these drawbacks, we developed a multigrid MALDI MSI technique (mMALDI MSI) that relies on automated inkjet printing to place differing matrices onto predefined dot grids. We used a cooled printhead to prevent cavitation of low viscosity solvents in the printhead nozzle. Improved spatial resolution of the dot grids was achieved by using a triple-pulse procedure that reduced droplet volume. The matrices can either be applied directly to the thaw-mounted tissue sample or by precoating the slide followed by mounting of the tissue sample. During the MALDI imaging process, we were able to precisely target different matrix point grids with the laser to simultaneously produce distinct mass spectra. Unlike the standard method, the prespotting approach optimizes the spectra quality, avoids analyte delocalization, and enables subsequent hematoxylin and eosin (H&E) staining.
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The authors thank Bruker Daltonics for its cooperation and Dr. Karl-Gunther Glowalla, (Friedrich Schiller University Jena, Experimental Service Unit) for providing the lab animals. This study was supported by the German Research Foundation [Deutsche Forschungsgemeinschaft (DFG)] Grant no. EG 102/4-1 and EG 102/5-1 as well as by the Carl-Zeiss Foundation (USS).
Conflict of interest
The authors declare that they incur no conflicts of interest.
Ulrich S. Schubert and Ferdinand von Eggeling contributed equally to this work.
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Urbanek, A., Hölzer, S., Knop, K. et al. Multigrid MALDI mass spectrometry imaging (mMALDI MSI). Anal Bioanal Chem 408, 3769–3781 (2016). https://doi.org/10.1007/s00216-016-9465-4
- Mass spectrometry imaging
- Inkjet printing
- Simultaneous analysis