Mr. William A. Smith Profile

William A. Smith

at Keele Univ

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Proceedings Article | 26 February 2014 Paper

Label free cell tracking in 3D tissue engineering constructs with high resolution imaging

W. Smith, K.-P. Lam, K. Dempsey, D. Mazzocchi-Jones, J. Richardson, Y. Yang

Proceedings Volume 8942, 894209 (2014) https://doi.org/10.1117/12.2039354

KEYWORDS: Nanofibers, 3D image processing, Phase contrast, Tissue engineering, Algorithm development, Microscopy, Neurons, Objectives, Image resolution, Microscopes

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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.

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