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.

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

Smith, W., Lam, K.-P., Dempsey, K., Mazzocchi-Jones, D., Richardson, J., & Yang, Y. Label free cell tracking in 3-D tissue engineering constructs with high resolution imaging. Presented at Dynamics and Fluctuations in Biomedical Photonics XI

• Presentation Conference Type: Conference Paper (published)
• Conference Name: Dynamics and Fluctuations in Biomedical Photonics XI
• Acceptance Date: Feb 24, 2014
• Publication Date: Feb 26, 2014
• Journal: Proceedings of SPIE
• Print ISSN: 0277-786X
• Electronic ISSN: 1996-756X
• 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
• Public URL: https://keele-repository.worktribe.com/output/409161
• Publisher URL: http://dx.doi.org/10.1117/12.2039354