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However, achieving high spatial resolution for both molecular classes remains challenging. Here, we combine tissue expansion with sequential imaging of lipids via matrix-assisted laser desorption\/ionization (MALDI) and intact proteins via nanospray desorption electrospray ionization (nano-DESI) on the same tissue section to achieve comprehensive high-resolution molecular maps. Physical stretching of the tissue on Parafilm \u201cM\u201d produced a 4.5-fold effective resolution enhancement in both the X and Y dimensions (a \u223c20-fold increase in area) in the mouse cerebellum for both MALDI and nano-DESI measurements. We further leveraged MALDI data to sharpen nano-DESI images using a partial least-squares (PLS) regression framework with a data consistency constraint. Finally, we applied midlevel fusion of MALDI and nano-DESI data to identify lipids and proteins localized to the same anatomical structures. Together, this straightforward workflow enables multimodal imaging of lipids and proteins from the same tissue section with improved spatial resolution and integrated analysis.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c02482","type":"journal-article","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T16:02:07Z","timestamp":1786723327000},"page":"24249-24258","source":"Crossref","is-referenced-by-count":0,"title":["Sequential\nMALDI and Nano-DESI MS Imaging of Lipids\nand Proteins on Stretched Tissue Samples"],"prefix":"10.1021","volume":"98","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3475-5981","authenticated-orcid":true,"given":"Timothy J.","family":"Trinklein","sequence":"first","affiliation":[{"name":"University of Illinois Urbana\u2212Champaign , , , ,","place":["Urbana, Illinois, United States, 61801-3681"]},{"name":"University of Illinois Urbana\u2212Champaign , , , ,","place":["Urbana, Illinois, United States, 61801-3681"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8464-3799","authenticated-orcid":true,"given":"Mushfeqa","family":"Iqfath","sequence":"additional","affiliation":[{"name":"Purdue University , , , ,","place":["West Lafayette, Indiana, United States, 47907-2050"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily A.","family":"Clayton","sequence":"additional","affiliation":[{"name":"University of Illinois Urbana\u2212Champaign , , , ,","place":["Urbana, Illinois, United States, 61801-3681"]},{"name":"University of Illinois Urbana\u2212Champaign , , , ,","place":["Urbana, Illinois, United States, 61801-3681"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0437-1493","authenticated-orcid":true,"given":"Stanislav S.","family":"Rubakhin","sequence":"additional","affiliation":[{"name":"University of Illinois Urbana\u2212Champaign , , , ,","place":["Urbana, Illinois, United States, 61801-3681"]},{"name":"University of Illinois Urbana\u2212Champaign , , , ,","place":["Urbana, Illinois, United States, 61801-3681"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4533-9644","authenticated-orcid":true,"given":"Julia","family":"Laskin","sequence":"additional","affiliation":[{"name":"Purdue University , , , ,","place":["West Lafayette, Indiana, United States, 47907-2050"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3107-9922","authenticated-orcid":true,"given":"Jonathan V.","family":"Sweedler","sequence":"additional","affiliation":[{"name":"University of Illinois Urbana\u2212Champaign , , , ,","place":["Urbana, Illinois, United States, 61801-3681"]},{"name":"University of Illinois Urbana\u2212Champaign , , , ,","place":["Urbana, Illinois, United States, 61801-3681"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,14]]},"reference":[{"issue":"4","key":"2026083118461609400_cit1","doi-asserted-by":"publisher","first-page":"2309","DOI":"10.1021\/cr3004295","article-title":"Analysis of Tissue Specimens by Matrix-Assisted Laser\nDesorption\/Ionization Imaging Mass Spectrometry in Biological and\nClinical Research","volume":"113","author":"Norris","year":"2013","journal-title":"Chem. Rev."},{"issue":"29","key":"2026083118461609400_cit2","doi-asserted-by":"publisher","first-page":"15517","DOI":"10.1021\/acs.analchem.4c05249","article-title":"Mass Spectrometry Imaging","volume":"97","author":"Ko\u0308rber","year":"2025","journal-title":"Anal.\nChem."},{"issue":"1","key":"2026083118461609400_cit3","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1021\/acs.analchem.5b04188","article-title":"Ambient Mass Spectrometry Imaging\nUsing Direct Liquid\nExtraction Techniques","volume":"88","author":"Laskin","year":"2016","journal-title":"Anal. Chem."},{"issue":"5","key":"2026083118461609400_cit4","doi-asserted-by":"publisher","first-page":"475","DOI":"10.1021\/acsmeasuresciau.4c00028","article-title":"Nanospray\nDesorption Electrospray Ionization Mass Spectrometry Imaging (Nano-DESI\nMSI): A Tutorial Review","volume":"4","author":"Iqfath","year":"2024","journal-title":"ACS Meas. Sci. 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