Features & Benefits of the Oris Cell Migration Assay

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1 Adherent cell migration without cell culture inserts The Oris Cell Migration Assay* is designed with a unique cell seeding stopper, detection mask, and stopper tool. There is no Transwell membrane insert needed. This unique plate design generates highly reproducible results using a microscope, digital imaging system or fluorescence plate reader. Simply adhere the seeded cells in each well, remove the stopper to create a detection zone, and measure the migration (see Figure 1: Assay Schematic). Oris assays are available with tissue culture treated plates, Collagen I coated plates, Fibronectin coated plates, or a TriCoated version that contains all 3 surfaces. Wound healing assays are normally performed using a scratch assay, which has high variability. The Oris Cell Migration Assay is an ideal alternative to the scratch assay since the cell seeding stoppers create a consistent detection zone to monitor 2-D closure. Features & Benefits of the Oris Cell Migration Assay Membrane-free migration - no cumbersome cell culture inserts; stain and view cells in the same well. Reproducible results - the unique design provides well-to-well CV's < 12% Preserve cell morphology - changes in cell structure can be monitored in real time Versatile - analyze data using multiple probes in a single well using a microscope, digital imager, or fluorescence plate reader Flexible - design kinetic or endpoint based cell migration assays without the use of special instrumentation

2 Sample Data Figure 2: Oris Cell Migration Analysis - Endpoint and Kinetic Detection: Wells were seeded with 50,000 HT-1080 cells. Cells adhered after 4 hours and the Oris stoppers were removed. All wells received CellTracker Green to fluorescently stain the cells. The assay was then incubated for 28 hours to permit cell migration and at various time points the fluorescence signals in the detection zones were measured using a plate reader. The images above illustrate representative data from pre-migration (t=0 hrs) and post-migration (t = 21 hrs) wells. The graph depicts a real-time analysis of cell migration that was prepared by transposing the fluorescent signal into cell numbers.

3 Figure 3: Oris Cell Migration Analysis - Inhibition: 3T3-Swiss albino fibroblast cells were pretreated with 5 ug/ml MMC to inhibit proliferation, and then 25,000 cells/well were seeded and allowed to adhere. Stoppers were then removed, Latrunculin A ( nm) was added to the wells, and the plate was incubated to permit cell migration. The cells were labeled with Calcein AM and the fluorescence quantified in the detection zone using a Synergy Bio-Tek plate reader. Latrunculin A inhibition of 3T3-Swiss albino cell migration was demonstrated by using the Oris Cell Migration Assay. The inhibitory effect of Latrunculin A was dose-dependent and complete inhibition of 3T3- Swiss albino cell migration was detected in the presence of at least 500 nm Latrunculin A.

4 ORDER the Oris Cell Migration Assays Product Number CM A Product Description Oris Cell Migration Assay, : Oris 96-well plate with Oris Cell Seeding Stoppers, 1 Package Size CM A Oris Cell Migration Assay, : Oris 96-well plates with Oris Cell Seeding Stoppers, 5 CM AC C Oris Cell Migration Assay - Collagen I Coated, : Oris Collagen I Coated, 96-well plate with Oris Cell Seeding Stoppers, 1 CM AC C Oris Cell Migration Assay - Collagen I Coated, : Oris Collagen I Coated, 96-well plates with Oris Cell Seeding Stoppers, 5 CM AF N Oris Cell Migration Assay - Fibronectin Coated, : Oris Fibronectin Coated, 96-well plate with Oris Cell Seeding Stoppers, 1 CM AF N Oris Cell Migration Assay - Fibronectin Coated, : Oris Fibronectin Coated, 96-well plates with Oris Cell Seeding Stoppers, 5 CM AT R Oris Cell Migration Assay - TriCoated, : Oris TriCoated (Tissue Culture Treated, Collagen I Coated, Fibronectin Coated), 96-well plate with Oris Cell Seeding Stoppers, 1 CM AT R Oris Cell Migration Assay - TriCoated, : Oris TriCoated (Tissue Culture Treated, Collagen I Coated, Fibronectin Coated), 96-well plates with Oris Cell Seeding Stoppers, 5 CM A. I R T Oris Stopper Tool 1 CM A. M M Oris Detection Mask 5

5 * Patent Pending. Oris is a trademark of Platypus Technologies, LLC CellTracker Green is a trademark of Invitrogen Corporation. Transwell is a registered trademark of Corning, Inc. Posters To view electronic versions of Platypus posters presented at the recent ASCB and AACR meetings go to this link