{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:18:04Z","timestamp":1785417484104,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,17]]},"group-title":"Biochemistry","reference-count":42,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T00:00:00Z","timestamp":1784246400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"award":["AI179891"],"award-info":[{"award-number":["AI179891"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,17]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Tuberculosis (TB) remains the world\u2019s leading infectious cause of death. Trehalose-based fluorogenic probes have emerged as powerful tools for labeling and studying mycobacteria, including\n                  <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                  (Mtb), the causative agent of tuberculosis. However, existing probes occupy a limited spectral range and require compromise between brightness, specificity, and functional readouts. Here, we report the design and characterization of two trehalose conjugates derived from Janelia Fluor\u00ae dyes, JF\n                  <jats:sub>635<\/jats:sub>\n                  -Tre and JF\n                  <jats:sub>646<\/jats:sub>\n                  -Tre, which extend the trehalose-based platform into the far-red region. Following NHS ester-mediated synthesis, the trehalose-conjugated analogs displayed strong far-red fluorescence, with excitation\/emission maxima at 638\/654 nm and 648\/663 nm for JF\n                  <jats:sub>635<\/jats:sub>\n                  -Tre and JF\n                  <jats:sub>646<\/jats:sub>\n                  -Tre, respectively. Both probes exhibited concentration- and time-dependent labeling of\n                  <jats:italic>Mycobacterium smegmatis<\/jats:italic>\n                  (Msmeg) and Mtb with minimal background fluorescence from the corresponding unconjugated dyes. Furthermore, we observed reduced labeling in heat-killed cells compared to live Msmeg, particularly for JF\n                  <jats:sub>646<\/jats:sub>\n                  -Tre, consistent with sensitivity to metabolic activity. Both JF-Tre derivatives produced significant cellular labeling and JF\n                  <jats:sub>635<\/jats:sub>\n                  -Tre distinguished untreated from INH-treated samples in drug susceptible Mtb, demonstrating the potential of the JF-Tre probes to report on INH susceptibility and resistance. Together, these findings expand the toolkit of trehalose-based probes and highlight how fluorophore identity influences probe performance in mycobacterial fluorescence imaging and drug susceptibility testing.\n                <\/jats:p>","DOI":"10.64898\/2026.07.16.737568","type":"posted-content","created":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T16:10:12Z","timestamp":1784304612000},"source":"Crossref","is-referenced-by-count":0,"title":["Expanding the palette of trehalose-based fluorophores for live mycobacterial detection"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-3084-0024","authenticated-orcid":false,"given":"Adriann","family":"Brodeth","sequence":"first","affiliation":[{"name":"UCLA Department of Bioengineering, Los Angeles, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-0185-3633","authenticated-orcid":false,"given":"Reena","family":"Dosanjh","sequence":"additional","affiliation":[{"name":"UCLA Department of Chemistry and Biochemistry, Los Angeles, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-3439-790X","authenticated-orcid":false,"given":"Lilith A.","family":"Schwartz","sequence":"additional","affiliation":[{"name":"UCLA Department of Chemistry and Biochemistry, Los Angeles, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6979-1655","authenticated-orcid":false,"given":"Sasha","family":"Kramer","sequence":"additional","affiliation":[{"name":"Bio-Techne (Tocris), The Watkins Building, Atlantic Road, Bristol, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7549-9446","authenticated-orcid":false,"given":"Sean","family":"Goggins","sequence":"additional","affiliation":[{"name":"Bio-Techne (Tocris), The Watkins Building, Atlantic Road, Bristol, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joel Cresser","family":"Brown","sequence":"additional","affiliation":[{"name":"Bio-Techne (Tocris), The Watkins Building, Atlantic Road, Bristol, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdulai","family":"Zigli","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, Central Michigan University, Mount Pleasant, MI, USA"},{"name":"Biochemistry, Cellular, and Molecular Biology Program, Central Michigan University, Mount Pleasant, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8402-359X","authenticated-orcid":false,"given":"Benjamin M.","family":"Swarts","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry, Central Michigan University, Mount Pleasant, MI, USA"},{"name":"Biochemistry, Cellular, and Molecular Biology Program, Central Michigan University, Mount Pleasant, MI, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1255-0803","authenticated-orcid":false,"given":"Mireille","family":"Kamariza","sequence":"additional","affiliation":[{"name":"UCLA Department of Bioengineering, Los Angeles, CA, USA"},{"name":"UCLA Molecular Biology Institute, Los Angeles, CA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026072010101022000_2026.07.16.737568v1.1","unstructured":"Global Tuberculosis Report 2024. 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