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

W. A. Smith; K.-P. Lam; K. P. Dempsey; D. Mazzocchi-Jones; J. B. Richardson; Y. Yang

doi:10.1117/12.2039354

26 February 2014 Label free cell tracking in 3D tissue engineering constructs with high resolution imaging

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

Proceedings Volume 8942, Dynamics and Fluctuations in Biomedical Photonics XI; 894209 (2014) https://doi.org/10.1117/12.2039354
Event: SPIE BiOS, 2014, San Francisco, California, United States

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.

Citation Download Citation

W. A. Smith, K.-P. Lam, K. P. Dempsey, D. Mazzocchi-Jones, J. B. Richardson, and Y. Yang "Label free cell tracking in 3D tissue engineering constructs with high resolution imaging", Proc. SPIE 8942, Dynamics and Fluctuations in Biomedical Photonics XI, 894209 (26 February 2014); https://doi.org/10.1117/12.2039354

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