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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.

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Authors

W.A. Smith

K.P. Dempsey

D. Mazzocchi-Jones

J.B. Richardson



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

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