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Optimizing Robustness of the Membrane-free, Oris™ Cell Migration Assay for High Throughput Screening using the BioTek Synergy™ HT Multi-Mode Microplate ReaderTélécharger
Related Products: Synergy HTX
February 02, 2009
Authors: Keren I. Hulkower, Renee L. Herber and Scott Gehler, Platypus Technologies, Madison; Paul Held, Ph.D and Xavier Amouretti, BioTek Instruments
Dysregulated cellular migration has been implicated in the failure of diabetic wound healing and in metastasis of cancer cells. Identification of potential therapeutic compounds that regulate cell motility would benefit from improved methods for high throughput screening (HTS).
The Oris™ Cell Migration Assay comprises a 96-well plate with silicone stoppers in each well that facilitate seeding cells annularly while excluding them from a 2 mm diameter centrally located detection zone. Following cell seeding and cell attachment, the stoppers are removed and cells migrate into the detection zone. Cells are then stained and an opaque mask, providing apertures that align with the detection zones, is attached to the bottom of the plate. The fluorescent signal is measured by using BioTek Instruments Synergy™ HT Multi-Mode Microplate Reader. Capture of fluorescence is limited to cells that have migrated into the detection zone based upon restrictive apertures of the opaque mask.
We demonstrate here the differential effects of (i) cell seeding on fibronectin, collagen I and tissue culture treated polystyrene substrates; (ii) staining with cytoplasmic, nuclear and cytoskeletal fluorescent dyes; and (iii) using a variety of mask aperture sizes on the robustness of the Oris™ Cell Migration Assay with HT-1080, MDA-MB-231 and NMuMG cells. Z-factors of > 0.5 were achieved for some combinations of these test parameters demonstrating the value of the Oris™ Cell Migration Assay for HTS applications.