Method for simultaneous tracking of thousands of unlabeled cells within a transparent 3D matrix.

Autor: Nette F; Fraunhofer Research and Development Center for Marine and Cellular Biotechnology, Lübeck, Germany., Guerra de Souza AC; Fraunhofer Research and Development Center for Marine and Cellular Biotechnology, Lübeck, Germany., Laskay T; Department of Infectious Diseases and Microbiology, University of Lübeck, Lübeck, Germany., Ohms M; Research Department Virus Immunology, Leibniz Institute for Experimental Virology, Hamburg, Germany., Dömer D; Department of Infectious Diseases and Microbiology, University of Lübeck, Lübeck, Germany., Drömann D; Medical Clinic III Pneumology, University Medical Center Schleswig-Holstein, Lübeck, Germany., Rapoport DH; Institute for Medical and Marine Biotechnology, University of Lübeck, Lübeck, Germany.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2022 Jun 24; Vol. 17 (6), pp. e0270456. Date of Electronic Publication: 2022 Jun 24 (Print Publication: 2022).
DOI: 10.1371/journal.pone.0270456
Abstrakt: 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.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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