{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T16:07:33Z","timestamp":1778774853403,"version":"3.51.4"},"posted":{"date-parts":[[2026,5,12]]},"group-title":"Microbiology","reference-count":37,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T00:00:00Z","timestamp":1778544000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["AI081704"],"award-info":[{"award-number":["AI081704"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["AI141893"],"award-info":[{"award-number":["AI141893"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["AI175177"],"award-info":[{"award-number":["AI175177"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["AI168222"],"award-info":[{"award-number":["AI168222"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Here, we develop CandiChrome, a multiplex labeling toolkit for\n                  <jats:italic>Candida albicans<\/jats:italic>\n                  , through combined\n                  <jats:italic>in vitro<\/jats:italic>\n                  and\n                  <jats:italic>in vivo<\/jats:italic>\n                  characterization of fluorescent proteins in a standard strain background. To this end, we screened 13 candidate fluorophores across the visible spectrum and assessed their practical performance based on brightness, stability, and usability. This analysis identified a seven-fluorophore set that achieved the most effective balance of signal strength, robustness, and compatibility. We used this optimized panel to build a modular multicolor platform that enables strain labeling, mixed-population imaging, and competition assays in\n                  <jats:italic>C. albicans<\/jats:italic>\n                  . This platform could resolve up to 21 distinct populations by flow cytometry and microscopy. Importantly, CandiChrome supported the resolution of differentially labeled populations both\n                  <jats:italic>in vitro<\/jats:italic>\n                  and in the murine host, supporting the simultaneous tracking of multiple strains in complex settings. Together, these results establish CandiChrome as a flexible platform for multiplex fungal imaging in a pathogenic species where multicolor tools remain underdeveloped.\n                <\/jats:p>","DOI":"10.64898\/2026.05.11.723596","type":"posted-content","created":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T06:35:12Z","timestamp":1778654112000},"source":"Crossref","is-referenced-by-count":0,"title":["A CandiChrome toolkit for multicolor labeling of\n                  <i>Candida<\/i>\n                  cells"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-2977-4842","authenticated-orcid":false,"given":"Cai-Ling","family":"Ke","sequence":"first","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Brown University, Providence, RI, 02912, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2084-2040","authenticated-orcid":false,"given":"Jingcheng","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Brown University, Providence, RI, 02912, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7660-6610","authenticated-orcid":false,"given":"Corey","family":"Frazer","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Brown University, Providence, RI, 02912, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7563-484X","authenticated-orcid":false,"given":"Richard J.","family":"Bennett","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology and Immunology, Brown University, Providence, RI, 02912, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026051408100461000_2026.05.11.723596v1.1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00038.2009"},{"key":"2026051408100461000_2026.05.11.723596v1.2","doi-asserted-by":"publisher","DOI":"10.1039\/b901966a"},{"key":"2026051408100461000_2026.05.11.723596v1.3","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201200408"},{"issue":"10","key":"2026051408100461000_2026.05.11.723596v1.4","first-page":"3437","article-title":"Near-infrared fluorescent probes: a next-generation tool for protein-labeling applications","volume":"12","year":"2020","journal-title":"Chem Sci"},{"key":"2026051408100461000_2026.05.11.723596v1.5","unstructured":"Tschanz, A. , D.Q. Matus , and I. Swinburne , Systematic comparison of mStayGold and mGold2 variants for live imaging in zebrafish. MicroPubl Biol, 2025. 2025."},{"key":"2026051408100461000_2026.05.11.723596v1.6","doi-asserted-by":"publisher","DOI":"10.1126\/science.1068539"},{"key":"2026051408100461000_2026.05.11.723596v1.7","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1037"},{"issue":"1","key":"2026051408100461000_2026.05.11.723596v1.8","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1038\/s41592-025-02962-2","article-title":"A highly photostable monomeric red fluorescent protein for dual-color 3D STED and time-lapse 3D SIM imaging","volume":"23","year":"2026","journal-title":"Nat Methods"},{"key":"2026051408100461000_2026.05.11.723596v1.9","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20081949"},{"issue":"11","key":"2026051408100461000_2026.05.11.723596v1.10","doi-asserted-by":"crossref","first-page":"1649","DOI":"10.1016\/j.bbamcr.2018.08.015","article-title":"Far-red acclimating cyanobacterium as versatile source for bright fluorescent biomarkers","volume":"1865","year":"2018","journal-title":"Biochim Biophys Acta Mol Cell Res"},{"key":"2026051408100461000_2026.05.11.723596v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3935"},{"key":"2026051408100461000_2026.05.11.723596v1.12","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-08050-8"},{"key":"2026051408100461000_2026.05.11.723596v1.13","doi-asserted-by":"publisher","DOI":"10.1002\/wdev.166"},{"key":"2026051408100461000_2026.05.11.723596v1.14","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-49455-y"},{"key":"2026051408100461000_2026.05.11.723596v1.15","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-38913-z"},{"key":"2026051408100461000_2026.05.11.723596v1.16","doi-asserted-by":"crossref","unstructured":"Zhu, J. , et al., A Validated Set of Fluorescent-Protein-Based Markers for Major Organelles in Yeast (Saccharomyces cerevisiae). mBio, 2019. 10(5).","DOI":"10.1128\/mBio.01691-19"},{"key":"2026051408100461000_2026.05.11.723596v1.17","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-143-2-303"},{"key":"2026051408100461000_2026.05.11.723596v1.18","doi-asserted-by":"publisher","DOI":"10.1002\/yea.1099"},{"key":"2026051408100461000_2026.05.11.723596v1.19","doi-asserted-by":"publisher","DOI":"10.1002\/yea.738"},{"key":"2026051408100461000_2026.05.11.723596v1.20","doi-asserted-by":"publisher","DOI":"10.1002\/yea.1674"},{"key":"2026051408100461000_2026.05.11.723596v1.21","doi-asserted-by":"publisher","DOI":"10.1128\/EC.00327-09"},{"key":"2026051408100461000_2026.05.11.723596v1.22","doi-asserted-by":"publisher","DOI":"10.1002\/cpmc.76"},{"key":"2026051408100461000_2026.05.11.723596v1.23","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-022-13214-0"},{"issue":"9","key":"2026051408100461000_2026.05.11.723596v1.24","doi-asserted-by":"crossref","first-page":"1412","DOI":"10.1038\/s41587-022-01469-x","article-title":"Author Correction: A highly photostable and bright green fluorescent protein","volume":"40","year":"2022","journal-title":"Nat Biotechnol"},{"key":"2026051408100461000_2026.05.11.723596v1.25","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4074"},{"key":"2026051408100461000_2026.05.11.723596v1.26","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-022-01467-6"},{"issue":"10","key":"2026051408100461000_2026.05.11.723596v1.27","doi-asserted-by":"crossref","first-page":"1608","DOI":"10.1016\/j.bbamcr.2019.06.018","article-title":"Design of small monomeric and highly bright near-infrared fluorescent proteins","volume":"1866","year":"2019","journal-title":"Biochim Biophys Acta Mol Cell Res"},{"key":"2026051408100461000_2026.05.11.723596v1.28","doi-asserted-by":"publisher","DOI":"10.1128\/mBio.02878-21"},{"key":"2026051408100461000_2026.05.11.723596v1.29","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.064097-0"},{"key":"2026051408100461000_2026.05.11.723596v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth819"},{"key":"2026051408100461000_2026.05.11.723596v1.31","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3891"},{"key":"2026051408100461000_2026.05.11.723596v1.32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0003647"},{"issue":"8","key":"2026051408100461000_2026.05.11.723596v1.33","doi-asserted-by":"crossref","first-page":"e1011810","DOI":"10.1371\/journal.pgen.1011810","article-title":"Gene dosage and protein valency impact phase separation and fungal cell fate","volume":"21","year":"2025","journal-title":"PLoS Genet"},{"key":"2026051408100461000_2026.05.11.723596v1.34","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.00780"},{"key":"2026051408100461000_2026.05.11.723596v1.35","doi-asserted-by":"publisher","DOI":"10.1016\/j.gene.2004.06.021"},{"key":"2026051408100461000_2026.05.11.723596v1.36","doi-asserted-by":"crossref","unstructured":"Lo, W.H. , et al., Synergistic Antifungal Activity of Chitosan with Fluconazole against Candida albicans, Candida tropicalis, and Fluconazole-Resistant Strains. Molecules, 2020. 25(21).","DOI":"10.3390\/molecules25215114"},{"key":"2026051408100461000_2026.05.11.723596v1.37","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2006-7-10-r100"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.05.11.723596","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T15:10:17Z","timestamp":1778771417000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.05.11.723596"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,12]]},"references-count":37,"URL":"https:\/\/doi.org\/10.64898\/2026.05.11.723596","relation":{},"subject":[],"published":{"date-parts":[[2026,5,12]]},"subtype":"preprint"}}