Nitric Oxide Imaging in Neurons Using Confocal Microscopy

The instability and low concentration of nitric oxide (NO) in specimens make it difficult to be detected directly. In this chapter, a method for imaging nitric oxide using laser scanning confocal microscopy (LSCM) is presented. A cultured hippocampal neuron is dyed with DAF-2 and observed under a Zeiss LSM 510 laser scanning confocal microscope. A 488-nm laser power is applied as excitation and the emission light from the labeled nitric oxide in the neuron is detected. In this way, nitric oxide is imaged and its intracellular kinetic change is monitored. Furthermore, image processing and visualization techniques are employed to help analyze the image data.