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Consistent, Repeatable Wound Healing Assays Incorporating a Novel, Automated Scratch Tool and Kinetic Label-Free ImagingDownload
February 06, 2019
Author: Brad Larson, BioTek Instruments, Inc., Winooski, VT, USA
Collective cell migration plays a role in numerous critical physiological processes, including morphogenesis and tissue regeneration. This movement has been shown to be critical in wound healing and cancer metastasis, therefore fully characterizing this activity is critical in identifying treatments for a variety of injury and disease states. One of the most widely used methods to measure collective cell migration is the wound healing or “scratch” assay. Following creation of a wound, or cell free zone, within the confluent cell layer, cell movement back into the wound area is monitored over time using cellular imaging. Kinetic and endpoint data then allow for quantification of cell migration, either when uninhibited or under the influence of a test molecule. For wound creation, commonly a pipette tip is manually dragged through the cells, which can lead to wounds that vary drastically in width, orientation, and in placement within the well. This yields increased variability in calculated measurements within replicate wells and across titrations, complicating final conclusions regarding the migratory ability of test cell models and treatments, especially when comparing assay-to-assay data. To increase the robustness of generated data, a consistent wound creation method is necessary.
Here we demonstrate use of a novel, automated tool to create scratch wounds in cell monolayers formed on the bottom of a microplate. With the single push of a button, and using a 4- or 8-pin head, consistent scratches of equivalent size and area are made in either 24- or 96-well plates. A multi-reservoir cleaning trough is incorporated on the deck of the tool. Using the onboard programmed procedure, unattended cleaning and decontamination of each pin is accomplished before and after use. This small tool fits in any size laminar flow hood, enabling wound creation in a sterile manner. Following washing, the plate can then be transferred to one of BioTek’s automated imagers or the BioSpa™ Live Cell Imaging System to kinetically monitor cell migration.