Method for simultaneous tracking of thousands of unlabeled cells within a transparent 3D matrix.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2022
2022
Historique:
received:
25
03
2022
accepted:
09
06
2022
entrez:
24
6
2022
pubmed:
25
6
2022
medline:
29
6
2022
Statut:
epublish
Résumé
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 (>50 mm2) at a time resolution of 4 seconds and a spatial resolution of ~1.5 μm in xy- and ~30 μm 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.
Identifiants
pubmed: 35749549
doi: 10.1371/journal.pone.0270456
pii: PONE-D-22-08894
pmc: PMC9232129
doi:
Substances chimiques
N-Formylmethionine Leucyl-Phenylalanine
59880-97-6
Collagen
9007-34-5
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0270456Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
Références
Technology (Singap World Sci). 2013 Oct 2;1(1):49
pubmed: 24809064
Mutat Res. 2013 Jan-Mar;752(1):10-24
pubmed: 22940039
Opt Express. 2020 Aug 31;28(18):26935-26952
pubmed: 32906958
Proc Natl Acad Sci U S A. 2012 Oct 2;109(40):16018-22
pubmed: 22988076
J Cell Sci. 2012 Jul 1;125(Pt 13):3015-24
pubmed: 22797912
Appl Opt. 2006 Jun 1;45(16):3893-901
pubmed: 16724155
Lab Chip. 2016 Aug 16;16(17):3304-16
pubmed: 27436197
J Biomed Opt. 2006 Sep-Oct;11(5):054032
pubmed: 17092181
Sci Adv. 2021 Jul 14;7(29):
pubmed: 34261650
Annu Rev Immunol. 2012;30:459-89
pubmed: 22224774
Opt Lett. 2003 Feb 1;28(3):164-6
pubmed: 12656319
PLoS One. 2010 Jul 30;5(7):e11921
pubmed: 20689600
Biomed Opt Express. 2016 Feb 23;7(3):949-62
pubmed: 27231600
Opt Lett. 2004 May 15;29(10):1132-4
pubmed: 15182009
Mol Oncol. 2007 Jun;1(1):84-96
pubmed: 18516279
Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11301-5
pubmed: 11572982
Front Immunol. 2019 Mar 29;10:635
pubmed: 30984197
Sci Signal. 2016 Dec 13;9(458):ra122
pubmed: 27965424
Nat Rev Immunol. 2013 Mar;13(3):159-75
pubmed: 23435331
Proc Natl Acad Sci U S A. 2007 Oct 30;104(44):17512-7
pubmed: 17959778
Nature. 2019 Apr;568(7753):546-550
pubmed: 30944468
Opt Lett. 2005 Mar 1;30(5):468-70
pubmed: 15789705
J Cell Biol. 1997 Dec 1;139(5):1349-60
pubmed: 9382879
J Biomed Opt. 2012 Dec;17(12):126018
pubmed: 23235893
Cytometry A. 2017 Jul;91(7):675-685
pubmed: 28544679
J Biomed Opt. 2009 Jan-Feb;14(1):014018
pubmed: 19256706
Nature. 1948 May 15;161(4098):777
pubmed: 18860291
Proc Natl Acad Sci U S A. 2005 May 10;102(19):6884-9
pubmed: 15860582
Assay Drug Dev Technol. 2014 May;12(4):207-18
pubmed: 24831787
Proc Natl Acad Sci U S A. 2014 Apr 22;111(16):5813-8
pubmed: 24711384
Phys Chem Chem Phys. 2016 May 14;18(18):12860-76
pubmed: 27104814
Dev Biol. 1983 Nov;100(1):64-119
pubmed: 6684600
Nat Immunol. 2008 Sep;9(9):960-9
pubmed: 18711433
Sensors (Basel). 2019 Mar 11;19(5):
pubmed: 30862101
Front Immunol. 2019 Mar 12;10:359
pubmed: 30915068
Sci Rep. 2018 Oct 31;8(1):16135
pubmed: 30382136
Cytometry A. 2017 May;91(5):433-442
pubmed: 28240818
Nat Med. 2010 Jun;16(6):708-12
pubmed: 20473304
Appl Opt. 2006 Feb 10;45(5):836-50
pubmed: 16512525
Nature. 2020 Jan;577(7791):537-542
pubmed: 31830756
Blood. 2012 Oct 4;120(14):e45-53
pubmed: 22915642
J Clin Periodontol. 2015 Jan;42(1):1-11
pubmed: 25360483
J Immunol. 2002 Jul 15;169(2):898-905
pubmed: 12097394
J Biol Chem. 1982 Jun 10;257(11):6280-6
pubmed: 6281264
J Appl Physiol (1985). 1990 Nov;69(5):1767-78
pubmed: 2272970
PLoS One. 2011;6(11):e27315
pubmed: 22087288