A high throughput, interactive imaging, bright‐field wound healing assay
Open Access
- 9 February 2011
- journal article
- research article
- Published by Wiley in Cytometry Part A
- Vol. 79A (3), 227-232
- https://doi.org/10.1002/cyto.a.21029
Abstract
The wound healing assay is a commonly used technique to measure cell motility and migration. Traditional methods of performing the wound healing assay suffer from low throughput and a lack of quantitative data analysis. We have developed a new method to perform a high‐throughput wound healing assay that produces quantitative data using the LEAP™ instrument. The LEAP™ instrument is used to create reproducible wounds in each well of a 96‐well plate by laser ablation. The LEAP™ then records bright field images of each well at several time points. A custom texture segmentation algorithm is used to determine the wound area of each well at each time point. This texture segmentation analysis can provide faster and more accurate image analysis than traditional methods. Experimental results show that reproducible wounds are created by laser ablation with a wound area that varies by less than 10%. This method was tested by confirming that neuregulin‐2β increases the rate of wound healing by MCF7 cells in a dose dependent manner. This automated wound healing assay has greatly improved the speed and accuracy, making it a suitable high‐throughput method for drug screening. © 2011 International Society for Advancement of CytometryKeywords
This publication has 15 references indexed in Scilit:
- Crossing barriers: The new dimension of 2D cell migration assaysJournal of Cellular Physiology, 2010
- Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblastsJournal of Ethnopharmacology, 2009
- The cancer stem cell marker CD133 has high prognostic impact but unknown functional relevance for the metastasis of human colon cancerThe Journal of Pathology, 2009
- Edge detection in microscopy images using curveletsBMC Bioinformatics, 2009
- Inter-conversion of neuregulin2 full and partial agonists for ErbB4Biochemical and Biophysical Research Communications, 2007
- High‐throughput laser‐mediated in situ cell purification with high purity and yieldCytometry Part A, 2004
- Neuregulin isoforms exhibit distinct patterns of ErbB family receptor activationOncogene, 2002
- In vitro endothelial wound repair. Interaction of cell migration and proliferation.Arteriosclerosis: An Official Journal of the American Heart Association, Inc., 1990
- The reorganization of microfilaments, centrosomes, and microtubules during in vitro small wound reendothelialization.The Journal of cell biology, 1988
- The initiation of cell division in a contact‐inhibited mammalian cell lineJournal of Cellular and Comparative Physiology, 1965