{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T18:06:40Z","timestamp":1789236400995,"version":"build-2803163510"},"posted":{"date-parts":[[2026,9,10]]},"group-title":"Biophysics","reference-count":14,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"UKRI Biotechnology and Biological Sciences Research Council","award":["BB\/T008369\/1"],"award-info":[{"award-number":["BB\/T008369\/1"]}]},{"award":["Nottingham Research Fellowship"],"award-info":[{"award-number":["Nottingham Research Fellowship"]}],"id":[{"id":"https:\/\/ror.org\/01ee9ar58","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Three-dimensional imaging of live\n                  <jats:italic>Caenorhabditis elegans<\/jats:italic>\n                  commonly relies on sequential z-stack acquisition, which can be slow and susceptible to movement and photobleaching. Here, we used light-field microscopy to capture dye and delivery-dependent fluorescence patterns across intact adult worms in a single exposure per channel. With a 40\u00d7\/1.20 NA water-immersion objective, each reconstructed dataset contained 121 axial slices spanning 109 \u00b5m at 0.91 \u00b5m spacing. In a representative adult, nematode signal extended from 40.88 to 85.4 \u00b5m, corresponding to approximately 44.5 \u00b5m of captured axial depth without sequential z scanning. Three fluorescence channels were acquired with a summed exposure time of 112 ms across a 685 \u00d7 1010 \u00b5m field of view. FM1-43 labelled intestinal, cuticular and vesicular structures, FM4-64 highlighted ingested bacteria and intestinal compartments, and Nile Red revealed vesicular and broader whole-organism fluorescence. Intrinsic blue and green autofluorescence signals were also resolved. These observations demonstrate rapid single-shot acquisition of whole-organism three-dimensional fluorescence information and show how delivery route influences the spatial distribution of commonly used fluorescent probes in\n                  <jats:italic>C. elegans<\/jats:italic>\n                  .\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Highlights<\/jats:title>\n                  <jats:list list-type=\"bullet\">\n                    <jats:list-item>\n                      <jats:p>Light-field microscopy captures whole-worm 3D fluorescence in a single exposure<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>A 109 \u00b5m axial range is rendered as 121 slices at 0.91 \u00b5m spacing<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>FM1-43, FM4-64 and Nile Red show distinct volumetric staining patterns<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>\n                        Delivery route alters fluorescence distribution across intact\n                        <jats:italic>C. elegans<\/jats:italic>\n                      <\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>Three-channel volumetric imaging uses a summed exposure time of 112 ms<\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                  <jats:fig id=\"ufig1\" position=\"float\" fig-type=\"figure\" orientation=\"portrait\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"749827v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.07.749827","type":"posted-content","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T21:35:15Z","timestamp":1789076115000},"source":"Crossref","is-referenced-by-count":0,"title":["Single-shot light-field microscopy captures delivery-dependent dye and autofluorescence patterns in\n                  <i>Caenorhabditis elegans<\/i>"],"prefix":"10.64898","author":[{"given":"Robert","family":"Markus","sequence":"first","affiliation":[{"name":"School of Life Sciences Imaging (SLIM), Biodiscovery Institute, University of Nottingham NG7 2UH, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward","family":"Rea","sequence":"additional","affiliation":[{"name":"ZEISS Research Microscopy Solutions, Cambourne, Cambridge, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nana A.","family":"Berfi","sequence":"additional","affiliation":[{"name":"School of Chemistry, University of Nottingham, Nottingham, NG7 2RD, UK"},{"name":"School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vincenzo","family":"Taresco","sequence":"additional","affiliation":[{"name":"School of Chemistry, University of Nottingham, Nottingham, NG7 2RD, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chris","family":"Hudson","sequence":"additional","affiliation":[{"name":"ZEISS Research Microscopy Solutions, Cambourne, Cambridge, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dominic R.A.","family":"White","sequence":"additional","affiliation":[{"name":"ZEISS Research Microscopy Solutions, Cambourne, Cambridge, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2067-4630","authenticated-orcid":false,"given":"Veeren M.","family":"Chauhan","sequence":"additional","affiliation":[{"name":"School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026091211000921000_2026.09.07.749827v1.1","doi-asserted-by":"publisher","DOI":"10.1145\/1141911.1141976"},{"key":"2026091211000921000_2026.09.07.749827v1.2","doi-asserted-by":"publisher","DOI":"10.1364\/OE.21.025418"},{"key":"2026091211000921000_2026.09.07.749827v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2964"},{"key":"2026091211000921000_2026.09.07.749827v1.4","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.12-02-00363.1992"},{"key":"2026091211000921000_2026.09.07.749827v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2009.10.002"},{"key":"2026091211000921000_2026.09.07.749827v1.6","doi-asserted-by":"publisher","DOI":"10.1051\/medsci\/20031967725"},{"key":"2026091211000921000_2026.09.07.749827v1.7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1001613"},{"key":"2026091211000921000_2026.09.07.749827v1.8","doi-asserted-by":"publisher","DOI":"10.18632\/aging.100936"},{"key":"2026091211000921000_2026.09.07.749827v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)81029-6"},{"key":"2026091211000921000_2026.09.07.749827v1.10","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.128.5.779"},{"key":"2026091211000921000_2026.09.07.749827v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-024-06126-4"},{"key":"2026091211000921000_2026.09.07.749827v1.12","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0912308107"},{"key":"2026091211000921000_2026.09.07.749827v1.13","unstructured":"Carl 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Available at: https:\/\/www.zeiss.com\/microscopy\/en\/products\/light-microscopes\/confocal-microscopes\/lightfield-4d.html (xaccessed 6 August 2026)."},{"key":"2026091211000921000_2026.09.07.749827v1.14","doi-asserted-by":"publisher","DOI":"10.1038\/s41592-019-0352-8"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.09.07.749827","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T18:00:18Z","timestamp":1789236018000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.09.07.749827"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,10]]},"references-count":14,"URL":"https:\/\/doi.org\/10.64898\/2026.09.07.749827","relation":{},"subject":[],"published":{"date-parts":[[2026,9,10]]},"subtype":"preprint"}}