Fluoview fv3000 filter confocal microscope system

Glass slides in a 24-well plate were seeded with about 50, 000 KB cells per well. After left to grow overnight, the cells were incubated with corresponding samples in medium (without FBS) at 37 °C for 1 h. After washing with PBS, the cells were fixed by 4% paraformaldehyde (PFA) and washed 3 times by PBS. The cytoskeleton of the fixed cells was treated with 0.1% Triton-×100 in PBS for 5 min to improve cell membrane permeability and then stained by Alexa Fluor 488 Phalloidin (Life Technologies) for 30 min at room temperature and then rinsed with PBS for 3 × 10 min. The cells were mounted with DAPI (Life Technologies). The slides were then observed under FluoView FV3000-Filter Confocal Microscope System (Olympus Corp.).

KB cells were grown on glass slides and cultured at 37 °C overnight. 100 nM Alexa Fluor 647 labeled G₄ RNA dendrimers were incubated with the cells at 37 °C for 30 min. After washing with PBS, the cells were fixed by 4% paraformaldehyde (PFA) and washed 3 times with PBS. The cytoskeleton and cell nucleus of the fixed cells were stained by Alexa Fluor 488 Phalloidin (Life Technologies) and Fluoroshield Mounting Medium with DAPI (Abcam), respectively. The cells were then assayed for cell binding by FluoView FV3000-Filter Confocal Microscope System (Olympus Corp).