{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T13:52:06Z","timestamp":1785937926809,"version":"3.56.0"},"reference-count":34,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["advanced.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Advanced Science"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Single\u2010microbe RNA sequencing (smRNA\u2010seq) emerges as a powerful approach for resolving microbial heterogeneity at single\u2010cell resolution, but current high\u2010throughput workflows still depend on chemical fixation to preserve cell\u2010of\u2010origin transcriptomic information during lysis, permeabilization, and downstream processing. Although fixation is integral to current smRNA\u2010seq workflows, its broader impact on overall performance remains unresolved because a practical alternative is lacking. Here, we present an smRNA\u2010seq strategy that integrates aqueous two\u2010phase system microcapsules with our previously developed smRandom\u2010seq method. Using polyethylene glycol diacrylate\u2010 and dextran\u2010based microcapsules, our method physically confines bacterial cells and released nucleic acids within individual microcompartments, thereby enabling single\u2010cell transcript capture in both fixation\u2010based and fixation\u2010free workflows. With two model species,\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    and\n                    <jats:italic>Acinetobacter baumannii<\/jats:italic>\n                    , we show that fixation\u2010free processing improves smRNA\u2010seq performance, increasing per\u2010cell gene detection sensitivity by approximately 50% relative to fixed controls while preserving similar overall transcriptional profiles. To our knowledge, this microcapsule\u2010integrated smRandom\u2010seq platform achieves the highest transcriptomic capture sensitivity among current high\u2010throughput smRNA\u2010seq platforms. This framework provides a practical alternative to fixation in high\u2010throughput smRNA\u2010seq and opens new prospects for the development of single\u2010microbe omics technologies.\n                  <\/jats:p>","DOI":"10.1002\/advs.77013","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T13:34:00Z","timestamp":1785936840000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Microcapsule\u2010Integrated smRandom\u2010seq Platform Enables Fixation\u2010Free Single Microbe RNA Sequencing"],"prefix":"10.1002","author":[{"given":"Mengdi","family":"Song","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine of the First Affiliated Hospital &amp; Liangzhu Laboratory Zhejiang University School of Medicine  Hangzhou 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China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1096-4706","authenticated-orcid":false,"given":"Zhengmin","family":"Tang","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine of the First Affiliated Hospital &amp; Liangzhu Laboratory Zhejiang University School of Medicine  Hangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2820-4243","authenticated-orcid":false,"given":"Yongcheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine of the First Affiliated Hospital &amp; Liangzhu Laboratory Zhejiang University School of Medicine  Hangzhou China"},{"name":"College of Biomedical Engineering and Instrument Science Zhejiang University  Hangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41564\u2010018\u20100201\u2010z"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41579\u2010019\u20100222\u20105"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.34133\/bdr.0052"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2011.10.004"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41422\u2010020\u20100332\u20107"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41392\u2010024\u201001946\u20106"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/d41586\u2010025\u201000075\u20106"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41564\u2010020\u20100729\u20106"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aba5257"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2023.01.002"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41564\u2010023\u201001348\u20104"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1093\/procel\/pwae027"},{"key":"e_1_2_11_14_1","doi-asserted-by":"crossref","DOI":"10.1002\/imt2.70035","article-title":"Single\u2010Microbe 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