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As the ROI approaches cellular dimensions (\u223c10\u00a0\u00b5m), laser\u2010induced photothermal damage and challenges in capturing microtissues in conventional LCM can compromise protein preservation and quantitative fidelity. This work introduces multi\u2010tiered \u00b5Dicers, fabricated by two\u2010photon polymerization, to mechanically dissect tissue slices into uniform microtissues down to 10\u00a0\u00b5m. The hierarchical blade architecture limits instantaneous blade\u2010tissue engagement and lowers the cutting force relative to single\u2010tier designs. For benchmarking, proteomic analysis is performed on ethanol\u2010fixed human squamous cell carcinoma microtissues generated by \u00b5Dicers and by LCM. Under identical Nanodroplet Processing in One pot for Trace Samples (nanoPOTS) and liquid chromatography\u2010mass spectrometry (LC\u2010MS) conditions, \u00b5Dicers yield more peptides and proteins than LCM, with the largest gains at 10\u201320\u00a0\u00b5m spatial resolution. Confocal imaging shows catapult\u2010associated cavities in LCM\u2010generated microtissues. This material loss, along with membrane\u2010limited protein extraction, likely reduces protein coverage. In contrast, multi\u2010tiered \u00b5Dicers enable reproducible microdissection down to 10\u00a0\u00b5m while maintaining high protein coverage. With the spatial registration of microtissues under development, \u00b5Dicers have the potential to complement LCM for next\u2010generation spatial proteomic workflows.<\/jats:p>","DOI":"10.1002\/smll.202514816","type":"journal-article","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T09:37:53Z","timestamp":1776245873000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Multi\u2010Tiered \u00b5Dicer Enables Protein\u2010Preserving Microdissection at 10\u00a0\u00b5m Resolution"],"prefix":"10.1002","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3264-4700","authenticated-orcid":false,"given":"Annatoma","family":"Arif","sequence":"first","affiliation":[{"name":"Department of Mechanical Engineering Stanford University  Stanford California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rashmi","family":"Kumar","sequence":"additional","affiliation":[{"name":"Environmental Molecular Sciences Laboratory Pacific Northwest National Laboratory  Richland Washington USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0523-6197","authenticated-orcid":false,"given":"Yumi","family":"Kwon","sequence":"additional","affiliation":[{"name":"Environmental Molecular Sciences Laboratory Pacific Northwest National Laboratory  Richland Washington USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramon","family":"Rodriguez","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Stanford University  Stanford California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seth Charles","family":"Cordts","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Stanford University  Stanford California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saisneha","family":"Koppaka","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Stanford University  Stanford California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Zhu","sequence":"additional","affiliation":[{"name":"Environmental Molecular Sciences Laboratory Pacific Northwest National Laboratory  Richland Washington USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ljiljana","family":"Pa\u0161a\u2010Toli\u0107","sequence":"additional","affiliation":[{"name":"Environmental Molecular Sciences Laboratory Pacific Northwest National Laboratory  Richland Washington USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1465-2432","authenticated-orcid":false,"given":"Sindy Kam Yan","family":"Tang","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering Stanford University  Stanford California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,15]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010018\u201003367\u2010w"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41587\u2010022\u201001302\u20105"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.202000077"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592\u2010024\u201002541\u2010x"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1101\/gr.5799207"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2024.108150"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xcrm.2022.100621"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3641"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010022\u201034824\u20102"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41592\u2010024\u201002454\u20109"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586\u2010025\u201009734\u20100"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0168\u20109525(98)01489\u20109"},{"key":"e_1_2_8_14_1","first-page":"1","article-title":"Beyond Laser Microdissection Technology: Follow the Yellow Brick Road for Cancer Research","volume":"4","author":"Legres L. 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