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Evaluation of Dynamic Cell Processes and Behavior Using Video Bioinformatics Tools

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Video Bioinformatics

Part of the book series: Computational Biology ((COBO,volume 22))

Abstract

Just as body language can reveal a person’s state of well-being, dynamic changes in cell behavior and morphology can be used to monitor processes in cultured cells. This chapter discusses how CL-Quant software, a commercially available video bioinformatics tool, can be used to extract quantitative data on: (1) growth/proliferation, (2) cell and colony migration, (3) reactive oxygen species (ROS) production, and (4) neural differentiation. Protocols created using CL-Quant were used to analyze both single cells and colonies. Time-lapse experiments in which different cell types were subjected to various chemical exposures were done using Nikon BioStations. Proliferation rate was measured in human embryonic stem cell colonies by quantifying colony area (pixels) and in single cells by measuring confluency (pixels). Colony and single cell migration were studied by measuring total displacement (distance between the starting and ending points) and total distance traveled by the colonies/cells. To quantify ROS production, cells were pre-loaded with MitoSOX Red™, a mitochondrial ROS (superoxide) indicator, treated with various chemicals, then total intensity of the red fluorescence was measured in each frame. Lastly, neural stem cells were incubated in differentiation medium for 12 days, and time lapse images were collected daily. Differentiation of neural stem cells was quantified using a protocol that detects young neurons. CL-Quant software can be used to evaluate biological processes in living cells, and the protocols developed in this project can be applied to basic research and toxicological studies, or to monitor quality control in culture facilities.

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Acknowledgments

Work presented in this chapter was supported by the following grants: TRDRP 22RT-0127, CIRM NE-A0005A-1E, NSF IGERT DGE 093667, a Cornelius Hopper Award from TRDRP, and a TRDRP postdoctoral fellowship 20FT-0084. We thank Dr. Evan Snyder for providing the mNSC and Randy Myers and Ned Jastromb for their help with the BioStations and CL-Quant.

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Authors and Affiliations

  1. UCR Stem Cell Center, Department of Cell Biology and Neuroscience, University of California, Riverside, 92521, USA

    Sabrina C. Lin, Henry Yip, Rattapol Phandthong, Barbara Davis & Prue Talbot

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  1. Sabrina C. Lin
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  2. Henry Yip
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  3. Rattapol Phandthong
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  5. Prue Talbot
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Correspondence to Prue Talbot .

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Editors and Affiliations

  1. University of California, Riverside, California, USA

    Bir Bhanu

  2. University of California, Riverside, California, USA

    Prue Talbot

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Lin, S.C., Yip, H., Phandthong, R., Davis, B., Talbot, P. (2015). Evaluation of Dynamic Cell Processes and Behavior Using Video Bioinformatics Tools. In: Bhanu, B., Talbot, P. (eds) Video Bioinformatics. Computational Biology, vol 22. Springer, Cham. https://doi.org/10.1007/978-3-319-23724-4_9

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  • DOI: https://doi.org/10.1007/978-3-319-23724-4_9

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  • Publisher Name: Springer, Cham

  • Print ISBN: 978-3-319-23723-7

  • Online ISBN: 978-3-319-23724-4

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