Nitric oxide (NO) and calcium ions (Ca2+) play critical role as molecular mediator in many physiological processes. However, their low concentration and instability in specimen make them difficult to be detected directly in neurons. We developed a method for imaging nitric oxide and calcium ions using Laser Scanning Confocal Microscopy (LSCM). Cultured hippocampal neuron is dyed and observed under Zeiss LSM 510 laser scanning confocal microscope. Excited by laser the emission light from the labeled nitric oxide and calcium ions in the neutron are detected. In this way, the nitric oxide and calcium ions are imaged and their intracellular kinetic change in monitored. Furthermore, image processing and visualization techniques are employed to help analyze the image data.