W.A. Smith
Label free cell tracking in 3-D tissue engineering constructs with high resolution imaging
Smith, W.A.; Lam, K.-P.; Dempsey, K.P.; Mazzocchi-Jones, D.; Richardson, J.B.; Yang, Y.
Authors
Ka-Po Lam k.p.lam@keele.ac.uk
K.P. Dempsey
D. Mazzocchi-Jones
J.B. Richardson
Ying Yang y.yang@keele.ac.uk
Abstract
Within the field of tissue engineering there is an emphasis on studying 3-D live tissue structures. Consequently, to investigate and identify cellular activities and phenotypes in a 3-D environment for all in vitro experiments, including shape, migration/proliferation and axon projection, it is necessary to adopt an optical imaging system that enables monitoring 3-D cellular activities and morphology through the thickness of the construct for an extended culture period without cell labeling. This paper describes a new 3-D tracking algorithm developed for Cell-IQ®, an automated cell imaging platform, which has been equipped with an environmental chamber optimized to enable capturing time-lapse sequences of live cell images over a long-term period without cell labeling. As an integral part of the algorithm, a novel auto-focusing procedure was developed for phase contrast microscopy equipped with 20x and 40x objectives, to provide a more accurate estimation of cell growth/trajectories by allowing 3-D voxels to be computed at high spatiotemporal resolution and cell density. A pilot study was carried out in a phantom system consisting of horizontally aligned nanofiber layers (with precise spacing between them), to mimic features well exemplified in cellular activities of neuronal growth in a 3-D environment. This was followed by detailed investigations concerning axonal projections and dendritic circuitry formation in a 3-D tissue engineering construct. Preliminary work on primary animal neuronal cells in response to chemoattractant and topographic cue within the scaffolds has produced encouraging results.
Journal Article Type | Conference Paper |
---|---|
Conference Name | Dynamics and Fluctuations in Biomedical Photonics XI |
Acceptance Date | Feb 24, 2014 |
Publication Date | Feb 26, 2014 |
Publicly Available Date | Mar 28, 2024 |
Journal | Proceedings of SPIE |
Print ISSN | 0277-786X |
Publisher | Society of Photo-optical Instrumentation Engineers |
Peer Reviewed | Peer Reviewed |
Series Title | Dynamics and Fluctuations in Biomedical Photonics XI |
DOI | https://doi.org/10.1117/12.2039354 |
Keywords | depth from focus, topographic surface, reconstruction, 2.5D imaging, multiscale, phase contrast microscopy, neuron proliferation |
Publisher URL | http://dx.doi.org/10.1117/12.2039354 |
Files
8942-11(1).pdf
(2.6 Mb)
PDF
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