Skip to main content
Article thumbnail
Location of Repository

An improved chamber for direct visualisation of chemotaxis

By D. Gullberg, A.J. Muinonen-Martin, D.M. Veltman, G. Kalna and R. Insall

Abstract

There has been a growing appreciation over the last decade that chemotaxis plays an important role in cancer migration, invasion and metastasis. Research into the field of cancer cell chemotaxis is still in its infancy and traditional investigative tools have been developed with other cell types and purposes in mind. Direct visualisation chambers are considered the gold standard for investigating the behaviour of cells migrating in a chemotactic gradient. We therefore drew up a list of key attributes that a chemotaxis chamber should have for investigating cancer cell chemotaxis. These include (1) compatibility with thin cover slips for optimal optical properties and to allow use of high numerical aperture (NA) oil immersion objectives; (2) gradients that are relatively stable for at least 24 hours due to the slow migration of cancer cells; (3) gradients of different steepnesses in a single experiment, with defined, consistent directions to avoid the need for complicated analysis; and (4) simple handling and disposability for use with medical samples. Here we describe and characterise the Insall chamber, a novel direct visualisation chamber. We use it to show GFP-lifeact transfected MV3 melanoma cells chemotaxing using a 60x high NA oil immersion objective, which cannot usually be done with other chemotaxis chambers. Linear gradients gave very efficient chemotaxis, contradicting earlier results suggesting that only polynomial gradients were effective. In conclusion, the chamber satisfies our design criteria, most importantly allowing high NA oil immersion microscopy to track chemotaxing cancer cells in detail over 24 hours

Publisher: Public Library of Science
Year: 2010
OAI identifier: oai:eprints.gla.ac.uk:49477
Provided by: Enlighten

Suggested articles

Citations

  1. (2010). A microfluidic imaging chamber for the direct observation of chemotactic transmigration.
  2. (1991). A new direct-viewing chemotaxis chamber.
  3. (2006). A parallel-gradient microfluidic chamber for quantitative analysis of breast cancer cell chemotaxis.
  4. (1977). Ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors.
  5. (1997). Analyzing chemotaxis using the Dunn directviewing chamber.
  6. (1999). Biophysical integration of effects of epidermal growth factor and fibronectin on fibroblast migration.
  7. (2007). Cell motility and cytoskeletal regulation in invasion and metastasis.
  8. (1981). Chemotactic reorientation of granulocytes stimulated with micropipettes containing fMet-Leu-Phe.
  9. (2007). Chemotaxis in shallow gradients is mediated independently of PtdIns 3-kinase by biased choices between random protrusions.
  10. (2009). Chemotaxis: finding the way forward with Dictyostelium.
  11. (2004). Chemotaxis: signalling the way forward.
  12. (2009). CircStat: A MATLAB Toolbox for Circular Statistics.
  13. (1998). Deconstructing (and reconstructing) cell migration.
  14. (1998). Epidermal growth factor alters fibroblast migration speed and directional persistence reciprocally and in a matrix-dependent manner.
  15. (1991). Establishment and characterization of a human melanoma cell line (MV3) which is highly metastatic in nude mice.
  16. (2008). Host nuclear factor-kappaB activation potentiates lung cancer metastasis.
  17. Insall RH (2006) Nap1 regulates Dictyostelium cell motility and adhesion through SCAR-dependent and -independent pathways.
  18. (1982). Measurement of chemotaxis in Boyden chamber filter assays. Is the checkerboard correction valid?
  19. (1997). Migration of highly aggressive MV3 melanoma cells in 3-dimensional collagen lattices results in local matrix reorganization and shedding of alpha2 and beta1 integrins and CD44.
  20. (2002). Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device.
  21. (2008). PDK1 regulates cancer cell motility by antagonising inhibition of ROCK1 by RhoE.
  22. (1998). Regulation of protrusion shape and adhesion to the substratum during chemotactic responses of mammalian carcinoma cells.
  23. (2002). Single cell behavior in metastatic primary mammary tumors correlated with gene expression patterns revealed by molecular profiling.
  24. (2005). The great escape: when cancer cells hijack the genes for chemotaxis and motility.
  25. (2000). Tumor invasion: role of growth factor-induced cell motility.
  26. (2010). Understanding eukaryotic chemotaxis: a pseudopod-centred view.

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.