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Conventional silane\u2010based methods suffer from poor stability, drastically hindering their reproducibility and performance. Here, we present an ethanol\u2010based aminopropyl\u2010silatrane functionalization protocol that achieves superior performance in a similar process timeframe. In particular, combining this approach with bacterial adhesin coupling, we demonstrated a significant twofold improvement in the fraction of attempted SMFS experiments yielding usable single\u2010molecule data. The method can be generally applied to silicon\u2010nitride and glass cantilevers and surfaces, including mechanically modified cantilevers optimized for fast time response, low stiffness, and low drift. Furthermore, we demonstrate a simple and cost\u2010efficient method to clean these functionalized cantilevers using common solvents and a benchtop UV\u2010ozone cleaner, allowing for refunctionalization and prolonged cantilever lifespans. Together, this allows for a reliable, reusable, and versatile functionalization protocol for high\u2010quality SMFS experiments.<\/jats:p>","DOI":"10.1002\/cbic.202500853","type":"journal-article","created":{"date-parts":[[2026,5,10]],"date-time":"2026-05-10T18:34:56Z","timestamp":1778438096000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Silanes to Silatranes: Robust Functionalization for Single\u2010Molecule Force Spectroscopy"],"prefix":"10.1002","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-0009-1726","authenticated-orcid":false,"given":"Thomas D.","family":"Courtney","sequence":"first","affiliation":[{"name":"Department of Chemistry Clemson University  Clemson South Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-2783-7525","authenticated-orcid":false,"given":"Christopher B.","family":"Hatchell","sequence":"additional","affiliation":[{"name":"Department of Chemistry Clemson University  Clemson South Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuliana","family":"O","sequence":"additional","affiliation":[{"name":"Department of Chemistry Clemson University  Clemson South Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0033-5874","authenticated-orcid":false,"given":"David R.","family":"Jacobson","sequence":"additional","affiliation":[{"name":"Department of Chemistry Clemson University  Clemson South Carolina USA"},{"name":"Medical Biophysics Graduate Program Clemson University  Clemson South Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,5,10]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201812072"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.1218"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/wnan.1253"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2021.167207"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.276.5315.1109"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-biophys-051013-022754"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1404446111"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2020083118"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.3934\/molsci.2015.3.190"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar2094"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmolb.2020.00085"},{"key":"e_1_2_7_13_1","first-page":"e50950","article-title":"Investigating Receptor\u2010ligand Systems of the Cellulosome with AFM\u2010based Single\u2010molecule Force Spectroscopy","author":"Jobst M. 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