Adding JF549 dyes to cells

Dr. Shao Chun Zen introduces the JF-549 dye single molecule tracking experiment and explains key requirements including working in darkness due to light sensitivity. The procedure will be demonstrated using HEX-293T cells expressing specific markers.

Add 1.5 ml of culture medium to a tube, then add 3.75 microliters of JF-549 dye stock solution. Ensure complete transfer of dye by pipetting up and down thoroughly within the medium.

Carefully remove the culture medium from the imaging disk, handling gently since cells attach loosely to the surface to avoid cell damage.

Thoroughly mix the dye-containing culture medium to ensure uniform concentration, then gently add the solution to the imaging disk containing the cells.

Cover the plate with clean aluminum foil to minimize light exposure, then incubate at 37 degrees Celsius for 15 minutes to allow dye labeling to occur.