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Three-Dimensional Reconstruction Imaging Method to Study the Function of EFHD2 in Invadopodia Formation

  • Yun Hyun Huh
  • Yu Ra Yeo
  • Woo Keun SongEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1929)

Abstract

Calcium and calcium-binding proteins play crucial roles in the regulation of actin dynamics, which contributes to cancer cell migration and invasion. In this chapter, we have focused on a three-dimensional imaging method to explore the pathophysiological function of EF-hand domain-containing protein D2 (EFHD2), a novel actin-binding protein. To overcome the limitations of two-dimensional imaging on substrate-coated cover glass for examination of invasive protrusions of cancer cells, we suggest three-dimensional reconstruction from optical z-sections of cells cultured on substrate-impregnated membrane filters of Transwell.

Key words

Calcium EFHD2 Actin Invadopodia 

Notes

Acknowledgments

This research was supported by the Bio Imaging and Cell Logistics Research Center (NRF-2016R1A5A1007318), Brain Research Program NRF-2014M3C7A1046050, NRF-2016M3C7A1905475, and NRF-2016R1C1B1013237 to YHH through the National Research Foundation of Korea funded by the Ministry of Science, ICT and Future Planning. The Bio Imaging and Cell Logistics Research Center provided access to microscopes.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Cell Logistics and Silver Health Research Center, School of Life Sciences, Gwangju Institute of Science and TechnologyGwangjuRepublic of Korea

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