Abstract
Thanks to a novel three-dimensional imaging platform based on lens-free microscopy, it is possible to perform multi-angle acquisitions and holographic reconstructions of 3D cell cultures directly into the incubator. Being able of reconstructing volumes as large as ~5 mm3 over a period of time covering several days, allows us to observe a broad range of migration strategies only present in 3D environment, whether it is single cell migration, collective migrations of cells and dispersal of cells. In addition we are able to distinguish new interesting phenomena, e.g. large-scale cell-to-matrix interactions (>1 mm), fusion of cell clusters into large aggregate (~10,000 µm2) and conversely, total dissociation of cell clusters into clumps of migrating cells. This work on a novel 3D + time lens-free microscopy technique thus expands the repertoire of phenomena that can be studied within 3D cell cultures.
Highlights
The imaging of 3D cell cultures opened a new window onto the study of many cellular processes as nicely reviewed in1. 3D + time imaging of 3D cell culture is usually performed through optical sectioning microscopy techniques, e.g. light-sheet microscopy and confocal live-cell microscopy
With the aim of developing such a gentle microscope, we developed a novel 3D + time lens-free microscope dedicated to the observation of dynamic biological processes present in 3D cell culture as previously presented in[3]
RWPE-1 cells have been used as a dynamic model of the signaling and interactions between organoids and mesenchyme that are required during organ development[7]
Summary
The imaging of 3D cell cultures opened a new window onto the study of many cellular processes as nicely reviewed in1. 3D + time imaging of 3D cell culture is usually performed through optical sectioning microscopy techniques, e.g. light-sheet microscopy and confocal live-cell microscopy. With the aim of developing such a gentle microscope, we developed a novel 3D + time lens-free microscope dedicated to the observation of dynamic biological processes present in 3D cell culture as previously presented in[3] It is based on the 3D lens-free microscopy setup introduced in[4] which enables a large angular coverage of the 3D scene thanks to its azimuthal acquisition geometry. Monitored the dynamic evolution of the extracellular matrix on a global scale and we were able to isolate the matrix deformations resulting from traction forces generated by large cell aggregates over long distances, up to 1.5 mm All these observations demonstrate that many important features of cell migration and cells-ECM (extra cellular matrix) interactions can be conveniently observed with our novel 3D + time lens-free microscope
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