{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,27]],"date-time":"2025-07-27T07:30:32Z","timestamp":1753601432921,"version":"3.37.3"},"reference-count":52,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2022,6,24]],"date-time":"2022-06-24T00:00:00Z","timestamp":1656028800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008530","name":"European Regional Development Fund","doi-asserted-by":"publisher","award":["123 17 024"],"award-info":[{"award-number":["123 17 024"]}],"id":[{"id":"10.13039\/501100008530","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Three-dimensional tracking of cells is one of the most powerful methods to investigate multicellular phenomena, such as ontogenesis, tumor formation or wound healing. However, 3D tracking in a biological environment usually requires fluorescent labeling of the cells and elaborate equipment, such as automated light sheet or confocal microscopy. Here we present a simple method for 3D tracking large numbers of unlabeled cells in a collagen matrix. Using a small lensless imaging setup, consisting of an LED and a photo sensor only, we were able to simultaneously track ~3000 human neutrophil granulocytes in a collagen droplet within an unusually large field of view (&gt;50 mm<jats:sup>2<\/jats:sup>) at a time resolution of 4 seconds and a spatial resolution of ~1.5 \u03bcm in xy- and ~30 \u03bcm in z-direction. The setup, which is small enough to fit into any conventional incubator, was used to investigate chemotaxis towards interleukin-8 (IL-8 or CXCL8) and N-formylmethionyl-leucyl-phenylalanine (fMLP). The influence of varying stiffness and pore size of the embedding collagen matrix could also be quantified. Furthermore, we demonstrate our setup to be capable of telling apart healthy neutrophils from those where a condition of inflammation was (I) induced by exposure to lipopolysaccharide (LPS) and (II) caused by a pre-existing asthma condition. Over the course of our experiments we have tracked more than 420.000 cells. The large cell numbers increase statistical relevance to not only quantify cellular behavior in research, but to make it suitable for future diagnostic applications, too.<\/jats:p>","DOI":"10.1371\/journal.pone.0270456","type":"journal-article","created":{"date-parts":[[2022,6,24]],"date-time":"2022-06-24T17:57:50Z","timestamp":1656093470000},"page":"e0270456","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Method for simultaneous tracking of thousands of unlabeled cells within a transparent 3D matrix"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8143-8597","authenticated-orcid":true,"given":"Falk","family":"Nette","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana Cristina","family":"Guerra de Souza","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tam\u00e1s","family":"Laskay","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mareike","family":"Ohms","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"D\u00f6mer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Dr\u00f6mann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2817-338X","authenticated-orcid":true,"given":"Daniel Hans","family":"Rapoport","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2022,6,24]]},"reference":[{"issue":"1","key":"pone.0270456.ref001","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/0012-1606(83)90201-4","article-title":"The embryonic cell lineage of the nematode Caenorhabditis elegans","volume":"100","author":"JE Sulston","year":"1983","journal-title":"Developmental Biology"},{"issue":"4","key":"pone.0270456.ref002","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1089\/adt.2014.573","article-title":"Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors","volume":"12","author":"R Edmondson","year":"2014","journal-title":"ASSAY and Drug Development Technologies"},{"issue":"7791","key":"pone.0270456.ref003","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1038\/s41586-019-1875-y","article-title":"A developmental landscape of 3D-cultured human pre-gastrulation embryos","volume":"577","author":"L Xiang","year":"2020","journal-title":"Nature"},{"key":"pone.0270456.ref004","doi-asserted-by":"crossref","first-page":"jcs.079509","DOI":"10.1242\/jcs.079509","article-title":"Deconstructing the third dimension\u2014how 3D culture microenvironments alter cellular cues","author":"BM Baker","year":"2012","journal-title":"Journal of Cell Science"},{"issue":"1","key":"pone.0270456.ref005","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.molonc.2007.02.004","article-title":"The morphologies of breast cancer cell lines in three-dimensional assays correlate with their profiles of gene expression","volume":"1","author":"PA Kenny","year":"2007","journal-title":"Molecular Oncology"},{"issue":"5","key":"pone.0270456.ref006","doi-asserted-by":"crossref","first-page":"836","DOI":"10.1364\/AO.45.000836","article-title":"Digital in-line holographic microscopy","volume":"45","author":"J Garcia-Sucerquia","year":"2006","journal-title":"Applied Optics"},{"issue":"4098","key":"pone.0270456.ref007","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1038\/161777a0","article-title":"A new microscopic principle","volume":"161","author":"D Gabor","year":"1948","journal-title":"Nature"},{"key":"pone.0270456.ref008","unstructured":"soft-matter\/trackpy: Trackpy v0.4.2; 2019. 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