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Opt. Express"],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:p>Immunostaining of thick biological tissues is often limited by slow antibody transport, resulting in long processing times and depth-dependent labeling. We describe an automated immunostaining approach based on an alternating bidirectional electric field, termed ARDEI, that improves labeling performance in millimeter-scale tissues. Under the applied electric-field conditions, tissue samples undergo reversible cyclic deformation, which may contribute to antibody redistribution by transiently altering tissue geometry. Using ARDEI, 1000 \u03bcm-thick mouse brain sections were labeled within 2.5 h, with improved signal penetration and spatial uniformity compared with passive diffusion and unidirectional electric-field staining under time-matched conditions. The system operates at 40 V under temperature-controlled conditions, minimizing thermal and structural perturbation. ARDEI supports three-dimensional imaging of neuronal and glial structures and can be applied to SHANEL-pretreated human brain tissue, where improved labeling uniformity was observed in 1000 \u03bcm-thick sections within 4 h. Under the tested conditions, repeated deformation cycles did not produce detectable structural distortion or signal degradation. These results show that ARDEI improves thick-tissue immunostaining performance and are consistent with a contribution from coupled electric-field-assisted transport and reversible tissue deformation. ARDEI provides a practical approach that can be integrated with existing optical imaging workflows for volumetric tissue analysis.<\/jats:p>","DOI":"10.1364\/boe.613433","type":"journal-article","created":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T18:00:09Z","timestamp":1787594409000},"page":"4901","update-policy":"https:\/\/doi.org\/10.1364\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Accelerated immunostaining of thick biological tissues using bidirectional electric fields with reversible deformation"],"prefix":"10.1364","volume":"17","author":[{"given":"Ziwen","family":"Zhou","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiao","family":"Xiao","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Nanhu Brain-computer Interface Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lijuan","family":"Du","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiazhu","family":"Zhu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenhong","family":"Du","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9095-5008","authenticated-orcid":true,"given":"Yao","family":"Zheng","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8328-4325","authenticated-orcid":true,"given":"Ke","family":"Si","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Gong","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a2xv884","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Nanhu Brain-computer Interface Institute"},{"name":"Zhejiang Institute of Medical Device Testing"},{"name":"Lingang Laboratory"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"285","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"key":"boe-17-9-4901-R1","doi-asserted-by":"publisher","first-page":"1756","DOI":"10.1038\/s41596-019-0160-8","type":"journal-article","volume":"14","author":"Dekkers","year":"2019","journal-title":"Nat. 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