A High Throughput, Interactive Imaging, Bright-Field Wound Healing Assay

Michael Zordan, Birck Nanotechnology Center, Purdue University
Christopher P. Mill, Purdue University; Auburn University
David J. Riese II, Purdue University; Auburn University
James F. Leary, Birck Nanotechnology Center, Purdue University

Date of this Version



Cytometry Part A: Volume 79A, Issue 3 March 2011, Pages 227–232


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 (TM) instrument. The LEAP (TM) instrument is used to create reproducible wounds in each well of a 96-well plate by laser ablation. The LEAP (TM) 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-2 beta 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. (C) 2011 International Society for Advancement of Cytometry


Nanoscience and Nanotechnology