% IMPORTANT: The following is UTF-8 encoded. This means that in the presence % of non-ASCII characters, it will not work with BibTeX 0.99 or older. % Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or % “biber”. @ARTICLE{Li:298584, author = {M. J. Li and L. C. Meyer and N. Meier and J. Witte and M. Maldacker and A. Seredynska$^*$ and J. Schueler and O. Schilling$^*$ and M. C. Föll$^*$}, title = {{S}patial {P}roteomics by {P}arallel {A}ccumulation-{S}erial {F}ragmentation {S}upported {MALDI} {MS}/{MS} {I}maging: {A} {F}irst {G}lance {I}nto {M}ultiplexed and {S}patial {P}eptide {I}dentification.}, journal = {Rapid communications in mass spectrometry}, volume = {39}, number = {9}, issn = {0951-4198}, address = {New York, NY}, publisher = {Wiley Interscience}, reportid = {DKFZ-2025-00301}, pages = {e10006}, year = {2025}, abstract = {In spatial proteomics, matrix-assisted laser desorption/ionization (MALDI) imaging enables rapid and cost-effective peptide measurements. Yet, in situ peptide identification remains challenging. Therefore, this study aims to integrate the trapped ion mobility spectrometry (TIMS)-based parallel accumulation-serial fragmentation (PASEF) into MALDI imaging of tryptic peptides to enable multiplexed MS/MS imaging.An initial MALDI TIMS MS1 survey measurement was performed, followed by a manual generation of a precursor list containing mass over charge values and ion mobility windows. Inside the dual TIMS system, submitted precursors were trapped, separately eluted by their ion mobility and analyzed in a quadrupole time-of-flight device, thereby enabling multiplexed MALDI MS/MS imaging. Finally, precursors were identified by peptide to spectrum matching.This study presents the first multiplexed MALDI TIMS MS/MS imaging (iprm-PASEF) of tryptic peptides. Its applicability was showcased on two histomorphologically distinct tissue specimens in a four-plex and five-plex setup. Precursors were successfully identified by the search engine MASCOT in one single MALDI imaging experiment for each respective tissue. Peptide identifications were corroborated by liquid-chromatography tandem mass spectrometry experiments and fragment colocalization analyses.In this study, we present a novel pipeline, based on iprm-PASEF, that allows the multiplexed and spatial identification of tryptic peptides in MALDI imaging. Hence, it marks a first step towards the integration of MALDI imaging into the emerging field of spatial proteomics.}, keywords = {Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization: methods / Proteomics: methods / Tandem Mass Spectrometry: methods / Animals / Peptides: chemistry / Peptides: analysis / Ion Mobility Spectrometry: methods / Peptide Fragments: analysis / Peptide Fragments: chemistry / MALDI imaging (Other) / MS/MS imaging (Other) / TIMS (Other) / iprm‐PASEF (Other) / peptide imaging (Other) / spatial proteomics (Other) / Peptides (NLM Chemicals) / Peptide Fragments (NLM Chemicals)}, cin = {FR01}, ddc = {530}, cid = {I:(DE-He78)FR01-20160331}, pnm = {899 - ohne Topic (POF4-899)}, pid = {G:(DE-HGF)POF4-899}, typ = {PUB:(DE-HGF)16}, pubmed = {pmid:39910729}, doi = {10.1002/rcm.10006}, url = {https://inrepo02.dkfz.de/record/298584}, }