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Live-cell confocal microscopy and quantitative 4D image analysis of anchor-cell invasion through the basement membrane in Caenorhabditis elegans.

Publication ,  Journal Article
Kelley, LC; Wang, Z; Hagedorn, EJ; Wang, L; Shen, W; Lei, S; Johnson, SA; Sherwood, DR
Published in: Nature protocols
October 2017

Cell invasion through basement membrane (BM) barriers is crucial in development, leukocyte trafficking and the spread of cancer. The mechanisms that direct invasion, despite their importance in normal and disease states, are poorly understood, largely because of the inability to visualize dynamic cell-BM interactions in vivo. This protocol describes multichannel time-lapse confocal imaging of anchor-cell invasion in live Caenorhabditis elegans. Methods presented include outline-slide preparation and worm growth synchronization (15 min), mounting (20 min), image acquisition (20-180 min), image processing (20 min) and quantitative analysis (variable timing). The acquired images enable direct measurement of invasive dynamics including formation of invadopodia and cell-membrane protrusions, and removal of BM. This protocol can be combined with genetic analysis, molecular-activity probes and optogenetic approaches to uncover the molecular mechanisms underlying cell invasion. These methods can also be readily adapted by any worm laboratory for real-time analysis of cell migration, BM turnover and cell-membrane dynamics.

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Published In

Nature protocols

DOI

EISSN

1750-2799

ISSN

1754-2189

Publication Date

October 2017

Volume

12

Issue

10

Start / End Page

2081 / 2096

Related Subject Headings

  • Time-Lapse Imaging
  • Microscopy, Confocal
  • Imaging, Three-Dimensional
  • Cell Membrane
  • Caenorhabditis elegans
  • Bioinformatics
  • Basement Membrane
  • Animals
  • 11 Medical and Health Sciences
  • 06 Biological Sciences
 

Citation

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Kelley, L. C., Wang, Z., Hagedorn, E. J., Wang, L., Shen, W., Lei, S., … Sherwood, D. R. (2017). Live-cell confocal microscopy and quantitative 4D image analysis of anchor-cell invasion through the basement membrane in Caenorhabditis elegans. Nature Protocols, 12(10), 2081–2096. https://doi.org/10.1038/nprot.2017.093
Kelley, Laura C., Zheng Wang, Elliott J. Hagedorn, Lin Wang, Wanqing Shen, Shijun Lei, Sam A. Johnson, and David R. Sherwood. “Live-cell confocal microscopy and quantitative 4D image analysis of anchor-cell invasion through the basement membrane in Caenorhabditis elegans.Nature Protocols 12, no. 10 (October 2017): 2081–96. https://doi.org/10.1038/nprot.2017.093.
Kelley LC, Wang Z, Hagedorn EJ, Wang L, Shen W, Lei S, et al. Live-cell confocal microscopy and quantitative 4D image analysis of anchor-cell invasion through the basement membrane in Caenorhabditis elegans. Nature protocols. 2017 Oct;12(10):2081–96.
Kelley, Laura C., et al. “Live-cell confocal microscopy and quantitative 4D image analysis of anchor-cell invasion through the basement membrane in Caenorhabditis elegans.Nature Protocols, vol. 12, no. 10, Oct. 2017, pp. 2081–96. Epmc, doi:10.1038/nprot.2017.093.
Kelley LC, Wang Z, Hagedorn EJ, Wang L, Shen W, Lei S, Johnson SA, Sherwood DR. Live-cell confocal microscopy and quantitative 4D image analysis of anchor-cell invasion through the basement membrane in Caenorhabditis elegans. Nature protocols. 2017 Oct;12(10):2081–2096.

Published In

Nature protocols

DOI

EISSN

1750-2799

ISSN

1754-2189

Publication Date

October 2017

Volume

12

Issue

10

Start / End Page

2081 / 2096

Related Subject Headings

  • Time-Lapse Imaging
  • Microscopy, Confocal
  • Imaging, Three-Dimensional
  • Cell Membrane
  • Caenorhabditis elegans
  • Bioinformatics
  • Basement Membrane
  • Animals
  • 11 Medical and Health Sciences
  • 06 Biological Sciences