Infinite m200 pro fluorescence plate reader

Transwell cell migration assays were performed using the CHEMICON^® QCM™ fluorimetric Cell Migration Assay Kit (Merck-Millipore, ECM 509), following the manufacturer’s instructions. Cells were serum starved for 24 h, then 300 μl of 5 × 10⁴ cells supplemented with 0,1% BSA were seeded into the migration chamber. A total of 500 μl of complete RPMI medium (with 10% FBS as a chemoattractant for LNCaP cells) was added to the feeder tray. The migration chamber plate was incubated in the cell detachment solution for 30 min at 37 °C, then 75 μl of Lysis Buffer/Dye Solution was added to the well and incubated for 15 min at RT, and fluorescence was measured with an Infinite M-200 Pro fluorescence plate reader (Tecan) using 480/520 nm a filter set. Transwell migration experiments were run in triplicates, quantified and reported graphically as bar charts. For the scratch motility experiments, cells were cultured to confluence in six-well plates at 37 °C. A 200 μl pipette tip was used to scratch the cells monolayer across the wells and images were captured using a Leica DFC 450 C microscope after the scratch (0 h) and after 24, 48, and 72 h. The migration rates of the LNCaP cells were estimated using ImageJ software (ImageJ 1.46r NIH). Experiments were performed in triplicates.