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P2055
High-Throughput Cell Migration Assay with the IsoCyte™ Laser Scanning Platform
Presenter Jayne Hesley, Blueshift Biotechnologies, USA
Additional Authors: Jayne Hesley, Brian Lee, Kui Lin, Steven C. Miller, Paul B. Comita, and Evan F. Cromwell
The biology of cell migration involves complex protein signaling mechanisms leading to changes in the cells cytoskeleton and ultimately to critical biological functions.  Assays that measure cell migration enable a more detailed understanding of the mechanisms involved and are important for drug and therapeutic discovery efforts.  One method commonly used to measure cell migration in the absence of a chemoattractant is the wound healing assay.  This assay consists of scratched or “wounded” cell monolayers and there are many challenges associated with performing this assay in high-throughput fashion; one major problem being the lack of uniformity of the wound area.  The Lin laboratory (Genentech) has demonstrated that cell migration in the PC-3 cell line is inhibited by specific drug and shRNA gene knockdown treatment.  Here we report on the use of the IsoCyte-DL™ (Blueshift Biotechnologies Inc.) laser scanning platform combined with the Oris™ cell migration assay kit (Platypus Technologies). The kit format consists of a 96-well plate where cell monolayers are formed in the presence of silicone inserts that mask only the center of each well.  After the inserts are removed, cell migration into the masked area is measured using the live cell fluorophore calcein-AM (Invitrogen) to label the cells for fluorescence detection. The IsoCyte™ scans whole well areas making it ideal for rapidly measuring cell migration within the uniform 2mm diameter unseeded region in the center of the well.  The results demonstrated an integrated image acquisition and image analysis process that enabled a simple, robust and high-throughput cell migration assay with the sensitivity required for the detection of drug or shRNA effect.