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Established differences in cell\u2010wall structure between Gram\u2010positive and Gram\u2010negative bacteria are expected to alter light scattering, motivating the hypothesis that the two groups differ in optical attenuation. This proof\u2010of\u2010concept study tested that optical prediction using optical coherence tomography. The optical attenuation coefficient was extracted from cross\u2010sectional images. Gram\u2010positive biofilms showed higher attenuation than Gram\u2010negative biofilms, with every cross\u2010group comparison significant, while species within each group were statistically indistinguishable, indicating a Gram\u2010level descriptor. Two deep\u2010learning classifiers separated the groups with over 97% accuracy because the attenuation distributions overlap minimally; this demonstrates the feasibility of automation rather than a requirement for deep learning. The attenuation differences are consistent with the hypothesis presented in the study, suggesting that these structural differences may contribute to the observed optical contrast.<\/jats:p>","DOI":"10.1002\/jbio.70372","type":"journal-article","created":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T00:26:31Z","timestamp":1791764791000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Optical Characterization of Gram\u2010Positive and Gram\u2010Negative Biofilms Using Swept\u2010Source Optical Coherence Tomography and Deep Learning"],"prefix":"10.1002","volume":"19","author":[{"given":"Nafisa","family":"Sultana","sequence":"first","affiliation":[{"name":"Biophotonics Lab, Department of Bioengineering &amp; Biotechnology Birla Institute of Technology\u2010Mesra  Ranchi India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abhishek","family":"Banerjee","sequence":"additional","affiliation":[{"name":"Biophotonics Lab, Department of Bioengineering &amp; Biotechnology Birla Institute of Technology\u2010Mesra  Ranchi India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Priyank","family":"Saxena","sequence":"additional","affiliation":[{"name":"Advanced Image Processing Lab, Department of Electronics &amp; Communication Engineering Birla Institute of Technology\u2010Mesra  Ranchi India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6945-9555","authenticated-orcid":false,"given":"Raju","family":"Poddar","sequence":"additional","affiliation":[{"name":"Biophotonics Lab, Department of Bioengineering &amp; Biotechnology Birla Institute of Technology\u2010Mesra  Ranchi India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,11]]},"reference":[{"key":"e_1_2_12_2_1","volume-title":"The Perfect Slime: Microbial Extracellular Polymeric Substances (EPS)","author":"Flemming H.\u2010C.","year":"2016"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsomega.0c02294"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.000032"},{"key":"e_1_2_12_5_1","first-page":"243","volume-title":"Springer Protocols Handbooks","author":"Mitra M.","year":"2021"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2019.00677"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00084-16"},{"key":"e_1_2_12_8_1","article-title":"High\u2010Resolution\u2010Three\u2010Dimensional\u2010Imaging\u2010of\u2010Biofilm\u2010Development\u2010Using\u2010Optical Coherence Tomogrpahy","volume":"11","author":"Xi C.","year":"2006","journal-title":"Journal of Biomedical Optics"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1117\/1.JMI.3.3.035507"},{"key":"e_1_2_12_10_1","first-page":"69","article-title":"Real\u2010Time Automated Thickness Measurement of the In\u00a0Vivo Human Tympanic Membrane Using Optical Coherence Tomography","volume":"5","author":"Hubler Z.","year":"2015","journal-title":"Quantitative Imaging in Medicine and Surgery"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.1117\/1.2435175"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.1088\/1555-6611\/ab1585"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/9781119246329.ch9"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/es9027937"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2015.01369"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0043-1354(94)90042-6"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.optcom.2025.131515"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.3390\/app8020184"},{"key":"e_1_2_12_19_1","unstructured":"M. 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