{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T16:59:03Z","timestamp":1791219543569,"version":"4.1.0"},"reference-count":27,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T00:00:00Z","timestamp":1791158400000},"content-version":"vor","delay-in-days":4,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T00:00:00Z","timestamp":1790812800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    We present a simple, low\u2010cost, and efficient protocol for fluorescent staining of\n                    <jats:italic>Bacillus<\/jats:italic>\n                    spp. endospores using the thioflavin T (ThT) dye. The article provides step\u2010by\u2010step procedures for culturing\n                    <jats:italic>Bacillus paranthracis<\/jats:italic>\n                    ,\n                    <jats:italic>Bacillus thuringiensis<\/jats:italic>\n                    (2 strains), and\n                    <jats:italic>Bacillus subtilis<\/jats:italic>\n                    , purifying endospores from pure cultures, and staining them with ThT. ThT\u2010stained endospores retain fluorescence for at least 2 years when stored at 4\u00b0C in the dark and remain fluorescent following formaldehyde fixation. Importantly, ThT staining does not impair spore viability, and stained endospores germinate normally. In contrast, vegetative\n                    <jats:italic>Bacillus<\/jats:italic>\n                    cells exhibit minimal to no ThT fluorescence. Overall, this robust and straightforward staining protocol is well suited for applications requiring fluorescently labeled endospores, including fluorescence microscopy, and provides a convenient plate\u2010based readout for germinant screening and related assays. \u00a9 2026 The Author(s).\n                    <jats:italic>Current Protocols<\/jats:italic>\n                    published by Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Endospore purification\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Fluorescent staining of endospores\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol<\/jats:bold>\n                    : Fluorescence microscopy of ThT\u2010stained spores\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70480","type":"journal-article","created":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T16:29:16Z","timestamp":1791217756000},"source":"Crossref","is-referenced-by-count":0,"title":["Purification and Thioflavin T Staining of\n                    <i>Bacillus<\/i>\n                    spp. Endospores for Fluorescence\u2010Based Assays"],"prefix":"10.1002","volume":"6","author":[{"given":"Ephrem Debebe","family":"Zegeye","sequence":"first","affiliation":[{"name":"Department of Paraclinical Sciences, Faculty of Veterinary Medicine Norwegian University of Life Sciences (NMBU)  \u00c5s Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Aspholm","sequence":"additional","affiliation":[{"name":"Department of Paraclinical Sciences, Faculty of Veterinary Medicine Norwegian University of Life Sciences (NMBU)  \u00c5s Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,5]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.00944\u201025"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10895\u2010014\u20101498\u20105"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01242\u201007"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780471729259.mca02as00"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro750"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/1462\u20102920.16678"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2005.06.006"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2025.1603957"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1093\/bbb\/zbac195"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.5560020312"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/mbio.02220\u201023"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574\u20106941.1995.tb00277.x"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574\u20106968.1996.tb08377.x"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2921"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.tbs\u20100014\u20102012"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/MMBR.64.3.548\u2010572.2000"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tim.2010.10.004"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2020106887"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.TBS\u20100003\u20102012"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010025\u201062896\u20103"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010024\u201051804\u2010w"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/mmbr.00050\u201015"},{"key":"e_1_2_8_24_1","first-page":"487","article-title":"Fluorescent stains, with special reference to amyloid and connective tissues","volume":"68","author":"Vassar P. 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