An improved chamber for direct visualisation of chemotaxis

Gullberg, D., Muinonen-Martin, A.J., Veltman, D.M., Kalna, G. and Insall, R. (2010) An improved chamber for direct visualisation of chemotaxis. PLoS ONE, 5(12), e15309. (doi:10.1371/journal.pone.0015309)

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Publisher's URL: http://dx.doi.org/10.1371/journal.pone.0015309

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.

Item Type:Articles
Additional Information:This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Insall, Professor Robert
Authors: Gullberg, D., Muinonen-Martin, A.J., Veltman, D.M., Kalna, G., and Insall, R.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
Journal Name:PLoS ONE
Publisher:Public Library of Science
ISSN:1932-6203
ISSN (Online):1932-6203
Published Online:01 January 2010
Copyright Holders:© 2010 Muinonen-Martin et al
First Published:First published in PLoS ONE 2010 5(12): e15309
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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