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Cytoskeleton Methods and Protocols pp 265–292Cite as

Quantitative Motion Analysis in Two and Three Dimensions

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1365))

Abstract

This chapter describes 2D quantitative methods for motion analysis as well as 3D motion analysis and reconstruction methods. Emphasis is placed on the analysis of dynamic cell shape changes that occur through extension and retraction of force generating structures such as pseudopodia and lamellipodia. Quantitative analysis of these structures is an underutilized tool in the field of cell migration. Our intent, therefore, is to present methods that we developed in an effort to elucidate mechanisms of basic cell motility, directed cell motion during chemotaxis, and metastasis. We hope to demonstrate how application of these methods can more clearly define alterations in motility that arise due to specific mutations or disease and hence, suggest mechanisms or pathways involved in normal cell crawling and treatment strategies in the case of disease. In addition, we present a 4D tumorigenesis model for high-resolution analysis of cancer cells from cell lines and human cancer tissue in a 3D matrix. Use of this model led to the discovery of the coalescence of cancer cell aggregates and unique cell behaviors not seen in normal cells or normal tissue. Graphic illustrations to visually display and quantify cell shape are presented along with algorithms and formulae for calculating select 2D and 3D motion analysis parameters.

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Acknowledgments

This work was supported by the Developmental Studies Hybridoma Bank (DSHB), a National Resource created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. We thank Brett Hanson, Joseph Ambrose, Kanoe Russell, Emma Buchele, Brian Kroll, Michele Livitz, Benjamin Soll, and Nicole Richardson for technical assistance. The monoclonal antibodies AIIB2, developed by C.H. Damsky, and P1B5, developed by E.A. Wayner and W.G. Carter, were obtained from the DSHB.

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Authors and Affiliations

  1. W.M. Keck Dynamic Image Analysis Facility, Department of Biological Sciences, University of Iowa, 302 BBE, Iowa City, IA, 52242, USA

    Deborah J. Wessels, Daniel F. Lusche, Spencer Kuhl, Amanda Scherer, Edward Voss & David R. Soll

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  1. Deborah J. Wessels
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  2. Daniel F. Lusche
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  3. Spencer Kuhl
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  4. Amanda Scherer
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  5. Edward Voss
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  6. David R. Soll
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Correspondence to David R. Soll .

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  1. Brooklyn College, Brooklyn, New York, USA

    Ray H. Gavin

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Wessels, D.J., Lusche, D.F., Kuhl, S., Scherer, A., Voss, E., Soll, D.R. (2016). Quantitative Motion Analysis in Two and Three Dimensions. In: Gavin, R. (eds) Cytoskeleton Methods and Protocols. Methods in Molecular Biology, vol 1365. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3124-8_14

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  • DOI: https://doi.org/10.1007/978-1-4939-3124-8_14

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