26 February 2014 Label free cell tracking in 3D tissue engineering constructs with high resolution imaging
Author Affiliations +
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.
© (2014) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
W. A. Smith, W. A. Smith, K.-P. Lam, K.-P. Lam, K. P. Dempsey, K. P. Dempsey, D. Mazzocchi-Jones, D. Mazzocchi-Jones, J. B. Richardson, J. B. Richardson, Y. Yang, 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); doi: 10.1117/12.2039354; https://doi.org/10.1117/12.2039354
PROCEEDINGS
8 PAGES


SHARE
Back to Top