{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,20]],"date-time":"2026-03-20T05:24:08Z","timestamp":1773984248012,"version":"3.50.1"},"reference-count":31,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T00:00:00Z","timestamp":1769558400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T00:00:00Z","timestamp":1769558400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Lasers Surg Med"],"published-print":{"date-parts":[[2026,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background and Objectives<\/jats:title>\n                    <jats:p>Accurate visualization of thermal damage zone (TDZ) in the dermis is critical for energy\u2010based device (EBD) and burn research. While nitroblue tetrazolium chloride (NBTC) staining and birefringence imaging are commonly used, both rely on detecting loss\u2010of\u2010signal, thereby only inferring TDZ indirectly. Other methods, such as hematoxylin and eosin and Masson's trichrome, are protocol\u2010sensitive and limited in sufficiency and consistency for TDZ visualization. This study aims to validate a novel staining protocol optimized for direct and selective visualization of thermally denatured collagen, termed Ahn\u2010van Gieson stain (AVG).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Protocol analysis was performed to identify sources of inconsistency in the iron hematoxylin\u2010based Verhoeff\u2010van Gieson stain, followed by comparison of multiple versions of modifications for optimization. Horizontal sections of the human dermis were irradiated with a defocused CO\u2082 laser beam under thermal recording with a 10.6\u202f\u03bcm notch filter. Thermal data were processed to generate peak temperature and Arrhenius integral maps for collagen denaturation. Slides stained with the optimized AVG protocol were overlaid with thermal maps for validation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Iron\u2010hematoxylin stained both TDZ and surrounding collagen regions, but subsequent differentiation using FeCl\n                      <jats:sub>3<\/jats:sub>\n                      led to partial loss of staining within TDZ, contributing to inconsistent results. Replacing the FeCl\n                      <jats:sub>3<\/jats:sub>\n                      differentiation step with 1\u2009min 1% acid alcohol treatment produced the most consistent and high\u2010contrast visualization of TDZ. The protocol required approximately 40\u2009min, functioned reliably on both frozen and paraffin\u2010embedded samples, and was compatible with birefringence imaging. The stained areas corresponded well with thermal maps for collagen denaturation, supporting the specificity of AVG for thermally denatured collagen.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>The novel protocol enables direct, high\u2010contrast, and reproducible visualization of thermally denatured collagen using standard equipment in a short time. It holds promise as a practical histological tool in EBD and burn research, where precise collagen damage visualization is essential.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/lsm.70101","type":"journal-article","created":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T05:43:14Z","timestamp":1769665394000},"page":"239-251","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An Optimized Staining Method for Visualization of Thermally Denatured Dermal Collagen"],"prefix":"10.1002","volume":"58","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5696-4699","authenticated-orcid":false,"given":"Ga Ram","family":"Ahn","sequence":"first","affiliation":[{"name":"Cutaneous Biology Research Center, Harvard Medical School Massachusetts General Hospital Boston Massachusetts USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9776-305X","authenticated-orcid":false,"given":"Sarfaraz Ahmed","family":"Quadri","sequence":"additional","affiliation":[{"name":"Cutaneous Biology Research Center, Harvard Medical School Massachusetts General Hospital Boston Massachusetts USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-9999-6760","authenticated-orcid":false,"given":"Heather M.","family":"Downs","sequence":"additional","affiliation":[{"name":"Cutaneous Biology Research Center, Harvard Medical School Massachusetts General Hospital Boston Massachusetts USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-5612-4044","authenticated-orcid":false,"given":"Parita","family":"T.Suwan","sequence":"additional","affiliation":[{"name":"Cutaneous Biology Research Center, Harvard Medical School Massachusetts General Hospital Boston Massachusetts USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Wang\u2010Evers","sequence":"additional","affiliation":[{"name":"Cutaneous Biology Research Center, Harvard Medical School Massachusetts General Hospital Boston Massachusetts USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John","family":"Warner\u2010Levy","sequence":"additional","affiliation":[{"name":"Cutaneous Biology Research Center, Harvard Medical School Massachusetts General Hospital Boston Massachusetts USA"},{"name":"The Faculty of Biology, Medicine and Health The University of Manchester Manchester UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4508-045X","authenticated-orcid":false,"given":"Dieter","family":"Manstein","sequence":"additional","affiliation":[{"name":"Cutaneous Biology Research Center, Harvard Medical School Massachusetts General Hospital Boston Massachusetts USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,1,28]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1097\/DSS.0000000000003397"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/jocd.16657"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1524-4725.1999.99091.x"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bjps.2012.04.028"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/jbio.201960024"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1088\/0967-3334\/35\/1\/55"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1117\/1.1644116"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2012.07.055"},{"key":"e_1_2_8_10_1","first-page":"289","article-title":"Recent Studies With the Electron Microscope on Ordered Aggregates of the Tropocollagen Molecules","volume":"1963","author":"Hodge A. 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