Advertisement

Measuring Chemotaxis Using Direct Visualization Microscope Chambers

  • Andrew J. Muinonen-Martin
  • David A. Knecht
  • Douwe M. Veltman
  • Peter A. Thomason
  • Gabriela Kalna
  • Robert H. Insall
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1046)

Abstract

Direct visualization chambers are considered the gold standard for measuring and analyzing chemotactic responses, because they allow detailed analysis of cellular behavior during the process of chemotaxis. We have previously described the Insall chamber, an improved chamber for measuring cancer cell chemotaxis. Here, we describe in detail how this system can be used to perform two key assays for both fast- and slow-moving mammalian and nonmammalian cell types. This allows for the detailed analysis of chemotactic responses in linear gradients at the levels of both overall cell behavior and subcellular dynamics.

Key words

Chemotaxis chamber Dictyostelium chemotaxis Hemopoietic cell chemotaxis Cancer cell chemotaxis Melanoma chemotaxis 

Notes

Acknowledgments

We thank Epigem (http://epigem.co.uk) for manufacturing the Insall chambers, Don MacBean for manufacturing the chamber holder cassettes and drilling the chambers, and Michael Carnell for writing the stitching plugin.

This work has been supported by a Wellcome Trust and a CRUK core grant.

References

  1. 1.
    Zigmond SH (1974) Mechanisms of sensing chemical gradients by polymorphonuclear leukocytes. Nature 249:450–452PubMedCrossRefGoogle Scholar
  2. 2.
    Roussos ET, Condeelis JS, Patsialou A (2011) Chemotaxis in cancer. Nat Rev Cancer 11:573–587PubMedCrossRefGoogle Scholar
  3. 3.
    Insall RH, Machesky LM (2009) Actin dynamics at the leading edge: from simple machinery to complex networks. Dev Cell 17:310–322PubMedCrossRefGoogle Scholar
  4. 4.
    Insall RH (2010) Understanding eukaryotic chemotaxis: a pseudopod-centred view. Nat Rev Mol Cell Biol 11:453–458PubMedCrossRefGoogle Scholar
  5. 5.
    Zigmond SH, Hirsch JG (1973) Leukocyte locomotion and chemotaxis. New methods for evaluation, and demonstration of a cell-derived chemotactic factor. J Exp Med 137:387–410PubMedCrossRefGoogle Scholar
  6. 6.
    Muinonen-Martin AJ, Veltman DM, Kalna G, Insall RH (2010) An improved chamber for direct visualisation of chemotaxis. PLoS One 5:e15309PubMedCrossRefGoogle Scholar
  7. 7.
    Soll DR (1995) The use of computers in understanding how animal cells crawl. Int Rev Cytol 163:43–104PubMedCrossRefGoogle Scholar
  8. 8.
    Zicha D, Dunn GA, Brown AF (1991) A new direct-viewing chemotaxis chamber. J Cell Sci 99:769–775PubMedGoogle Scholar
  9. 9.
    Berens P (2009) CircStat: a MATLAB toolbox for circular statistics. J Stat Software 31:1–21Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Andrew J. Muinonen-Martin
    • 1
  • David A. Knecht
    • 2
  • Douwe M. Veltman
    • 1
  • Peter A. Thomason
    • 1
  • Gabriela Kalna
    • 1
  • Robert H. Insall
    • 1
  1. 1.CRUK Beatson Institute for Cancer ResearchGlasgowUK
  2. 2.Department of Molecular and Cell BiologyUniversity of ConnecticutStorrsUSA

Personalised recommendations