Co-localization of GFP-tagged Proteins with Markers of Cellular Components and Live Cell Imaging

  1. Dilute the marker (e.g., Lysotracker, mitotracker, or Golgi marker) in serum-free HEPES-MEM in a tube and mix it well. (1-2 uM final concentration; 1.5 - 2 ml per 35 mm dish)
  2. Gently rinse the cells after transfection with serum-free HEPES-MEM and add the medium with the markers into the plate carefully.
  3. Incubate the culture for 20-25 minutes at room temperature.
  4. Change the medium (rinse) the cells with 5 ml HEPES-MEM once to remove the dye solution. Add 6-7 ml of the same medium to the dish.
  5. Examine the data under the confocal microscope and collect the images using the FITC/CY3/Phase program.