000283220 001__ 283220 000283220 005__ 20260116093310.0 000283220 0247_ $$2doi$$a10.1021/acs.analchem.5c05199 000283220 0247_ $$2pmid$$apmid:41408757 000283220 0247_ $$2ISSN$$a0003-2700 000283220 0247_ $$2ISSN$$a0096-4484 000283220 0247_ $$2ISSN$$a1520-6882 000283220 0247_ $$2ISSN$$a1541-4655 000283220 037__ $$aDZNE-2026-00069 000283220 041__ $$aEnglish 000283220 082__ $$a540 000283220 1001_ $$00009-0004-6998-4690$$aSidorov, Iulia$$b0 000283220 245__ $$aIn-Source Fragmentation Annotation in Sterol Mass Spectrometry Imaging. 000283220 260__ $$aColumbus, Ohio$$bAmerican Chemical Society$$c2026 000283220 3367_ $$2DRIVER$$aarticle 000283220 3367_ $$2DataCite$$aOutput Types/Journal article 000283220 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article$$bjournal$$mjournal$$s1768552242_4731 000283220 3367_ $$2BibTeX$$aARTICLE 000283220 3367_ $$2ORCID$$aJOURNAL_ARTICLE 000283220 3367_ $$00$$2EndNote$$aJournal Article 000283220 520__ $$aSpatial biology has emerged as a pivotal area in many life science fields, with mass spectrometry imaging (MSI) becoming a cornerstone for molecular imaging. Among recent advancements to increase sensitivity, MALDI-2 technology has significantly expanded the molecular space accessible to MSI, increasing the ion yields of neutral metabolites, such as sterols. Sterols have recently taken center stage in numerous (patho-) physiological processes, including neurodegenerative diseases that have attracted significant scientific interest. However, in-source fragmentation (ISF) poses a substantial challenge for accurate biological interpretation of mass spectrometric data. In this study, we observed and investigated the ISF of cholesterol during MSI under MALDI and MALDI-2 conditions. Using a murine intervention model, we demonstrate how ISF can compromise the accuracy of biological interpretations, potentially leading to significant misinterpretations. Our study underscores the critical need to address ISF to ensure accurate molecular annotation, particularly through tandem mass spectrometry of in-source fragments. This is especially important when using MALDI-2 techniques. Furthermore, we introduce a high-resolution (5 μm) MSI technique, enabling the precise spatial analysis of cholesterol distribution. 000283220 536__ $$0G:(DE-HGF)POF4-351$$a351 - Brain Function (POF4-351)$$cPOF4-351$$fPOF IV$$x0 000283220 588__ $$aDataset connected to CrossRef, PubMed, , Journals: pub.dzne.de 000283220 650_7 $$097C5T2UQ7J$$2NLM Chemicals$$aCholesterol 000283220 650_7 $$2NLM Chemicals$$aSterols 000283220 650_2 $$2MeSH$$aAnimals 000283220 650_2 $$2MeSH$$aSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization: methods 000283220 650_2 $$2MeSH$$aMice 000283220 650_2 $$2MeSH$$aCholesterol: analysis 000283220 650_2 $$2MeSH$$aCholesterol: metabolism 000283220 650_2 $$2MeSH$$aSterols: analysis 000283220 650_2 $$2MeSH$$aSterols: chemistry 000283220 650_2 $$2MeSH$$aMolecular Imaging: methods 000283220 7001_ $$00000-0002-0773-0095$$aOgrinc, Nina$$b1 000283220 7001_ $$0P:(DE-2719)9002306$$aSpieth, Lena$$b2$$udzne 000283220 7001_ $$aKopcil, Michal$$b3 000283220 7001_ $$aHeijink, Marieke$$b4 000283220 7001_ $$aDalebout, Hans$$b5 000283220 7001_ $$aSaher, Gesine$$b6 000283220 7001_ $$0P:(DE-2719)9001700$$aBerghoff, Stefan A$$b7$$udzne 000283220 7001_ $$00000-0002-4749-0014$$aSiuzdak, Gary$$b8 000283220 7001_ $$00000-0003-1684-1894$$aGiera, Martin$$b9 000283220 773__ $$0PERI:(DE-600)1483443-1$$a10.1021/acs.analchem.5c05199$$gVol. 98, no. 1, p. 509 - 518$$n1$$p509 - 518$$tAnalytical chemistry$$v98$$x0003-2700$$y2026 000283220 8564_ $$uhttps://pub.dzne.de/record/283220/files/DZNE-2026-00069_Restricted.pdf 000283220 8564_ $$uhttps://pub.dzne.de/record/283220/files/DZNE-2026-00069_Restricted.pdf?subformat=pdfa$$xpdfa 000283220 9101_ $$0I:(DE-588)1065079516$$6P:(DE-2719)9002306$$aDeutsches Zentrum für Neurodegenerative Erkrankungen$$b2$$kDZNE 000283220 9101_ $$0I:(DE-588)1065079516$$6P:(DE-2719)9001700$$aDeutsches Zentrum für Neurodegenerative Erkrankungen$$b7$$kDZNE 000283220 9131_ $$0G:(DE-HGF)POF4-351$$1G:(DE-HGF)POF4-350$$2G:(DE-HGF)POF4-300$$3G:(DE-HGF)POF4$$4G:(DE-HGF)POF$$aDE-HGF$$bGesundheit$$lNeurodegenerative Diseases$$vBrain Function$$x0 000283220 915__ $$0StatID:(DE-HGF)0420$$2StatID$$aNationallizenz$$d2024-12-09$$wger 000283220 915__ $$0StatID:(DE-HGF)0200$$2StatID$$aDBCoverage$$bSCOPUS$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)0300$$2StatID$$aDBCoverage$$bMedline$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)0199$$2StatID$$aDBCoverage$$bClarivate Analytics Master Journal List$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)1150$$2StatID$$aDBCoverage$$bCurrent Contents - Physical, Chemical and Earth Sciences$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)0160$$2StatID$$aDBCoverage$$bEssential Science Indicators$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)1030$$2StatID$$aDBCoverage$$bCurrent Contents - Life Sciences$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)0113$$2StatID$$aWoS$$bScience Citation Index Expanded$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)0150$$2StatID$$aDBCoverage$$bWeb of Science Core Collection$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)0100$$2StatID$$aJCR$$bANAL CHEM : 2022$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)0600$$2StatID$$aDBCoverage$$bEbsco Academic Search$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)0030$$2StatID$$aPeer Review$$bASC$$d2024-12-09 000283220 915__ $$0StatID:(DE-HGF)9905$$2StatID$$aIF >= 5$$bANAL CHEM : 2022$$d2024-12-09 000283220 9201_ $$0I:(DE-2719)1110008$$kAG Simons$$lMolecular Neurobiology$$x0 000283220 980__ $$ajournal 000283220 980__ $$aEDITORS 000283220 980__ $$aVDBINPRINT 000283220 980__ $$aI:(DE-2719)1110008 000283220 980__ $$aUNRESTRICTED