Transfection – 18/07/2011 Data collection and analysis – 20/07/11 HeLa Cell Transfection Experiment 96 well plate, 10,000 cells per well 10,000 HeLa cells were seeded in each of the highlighted wells, 24hours before transfection, allowing the cells to properly attach to the plate. 24hours after seeding complexes of DNA and nanoparticle were formed to the following ratios: 1. nTMag 0.1ul – GFP 0.1ul 2. OzBio 0.1ul – GFP 0.1ul 3. nTMag 0.1ul – LUX 0.1ul 4. OzBio 0.1ul – LUX 0.1ul The DNA and nanoparticles were allowed to complex for 15mins at room temperature. After the DNA and nanoparticles had complexed, the cell culture medium was aspirated and 100ul of new medium (-FCS) and nanoparticle/DNA complex were added to the appropriate wells, as outlined in the diagram below: After the medium and complexes had been added to wells, the plate was placed on a magnefect-nano 96-well oscillating magnet array for 30mins, in an incubator (37ºC,5% CO 2 ). The magnefect-nano system was set at 2Hz frequency, 30min duration and 0.2mm displacement. This process caused the nanoparticle/DNA complexes to be drawn down onto the cells. The oscillation increased the agitation of the nanoparticles onto the surface of the cells, encouraging the cells to take in the nanoparticle/DNA complexes via endocytosis. The hypothesis was that 48hours later, the cells would have transcribed the DNA and translated it into a protein. In the case of GFP, the cells with the protein would glow green under a fluorescent microscope, and the cells that were transfected with LUX DNA would produce light (luminescence). After the 30mins on the magnet array, the cells were taken out of the incubator and off the magnet array. The wells were then aspirated of medium and nanoparticles, so as to make sure the transfection occurred over 30mins only. 100ul of new (+FCS) medium was then added to the wells, which were left for 48hours in an incubator at 37º,5% CO 2 . nTMag 0.1ul – GFP DNA 0.1ug OzBio 0.1ul – GFP DNA 0.1ug nTMag 0.1ul – LUX DNA 0.1ug OzBio 0.1ul – LUX DNA 0.1ug