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Proceedings Paper

Label free cell tracking in 3D tissue engineering constructs with high resolution imaging
Author(s): W. A. Smith; K.-P. Lam; K. P. Dempsey; D. Mazzocchi-Jones; J. B. Richardson; Y. Yang
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Paper 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.

Paper Details

Date Published: 26 February 2014
PDF: 8 pages
Proc. SPIE 8942, Dynamics and Fluctuations in Biomedical Photonics XI, 894209 (26 February 2014); doi: 10.1117/12.2039354
Show Author Affiliations
W. A. Smith, Keele Univ. (United Kingdom)
K.-P. Lam, Keele Univ. (United Kingdom)
K. P. Dempsey, Keele Univ. (United Kingdom)
D. Mazzocchi-Jones, Institute for Science & Technology in Medicine (United Kingdom)
J. B. Richardson, The Robert Jones and Agnes Hunt Orthopaedic Hospital NHS Foundation Trust (United Kingdom)
Y. Yang, Institute for Science & Technology in Medicine (United Kingdom)

Published in SPIE Proceedings Vol. 8942:
Dynamics and Fluctuations in Biomedical Photonics XI
Valery V. Tuchin; Donald D. Duncan; Kirill V. Larin; Martin J. Leahy; Ruikang K. Wang, Editor(s)

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