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Open Access Highly Accessed Methodology article

A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods

Justin C Yarrow12, Zachary E Perlman12, Nicholas J Westwood23 and Timothy J Mitchison12*

Author Affiliations

1 Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA

2 The Institute of Chemistry and Cell Biology (ICCB), Harvard Medical School, Boston, MA 02115, USA

3 School of Chemistry and Centre for Biomolecular Sciences, University of St Andrews, North Haugh, St Andrews, UK

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BMC Biotechnology 2004, 4:21  doi:10.1186/1472-6750-4-21

Published: 9 September 2004

Abstract

Background

Cell migration is a complex phenomenon that requires the coordination of numerous cellular processes. Investigation of cell migration and its underlying biology is of interest to basic scientists and those in search of therapeutics. Current migration assays for screening small molecules, siRNAs, or other perturbations are difficult to perform in parallel at the scale required to screen large libraries.

Results

We have adapted the commonly used scratch wound healing assay of tissue-culture cell monolayers to a 384 well plate format. By mechanically scratching the cell substrate with a pin array, we are able to create characteristically sized wounds in all wells of a 384 well plate. Imaging of the healing wounds with an automated fluorescence microscope allows us to distinguish perturbations that affect cell migration, morphology, and division. Readout requires ~1 hr per plate but is high in information content i.e. high content. We compare readouts using different imaging technologies, automated microscopy, scanners and a fluorescence macroscope, and evaluate the trade-off between information content and data acquisition rate.

Conclusions

The adaptation of a wound healing assay to a 384 well format facilitates the study of aspects of cell migration, tissue reorganization, cell division, and other processes that underlie wound healing. This assay allows greater than 10,000 perturbations to be screened per day with a quantitative, high-content readout, and can also be used to characterize small numbers of perturbations in detail.