Methods in Molecular Biology Volume 913, 2012, pp 191-200

In vivo Imaging of Nitric Oxide and Reactive Oxygen Species Using Laser Scanning Confocal Microscopy

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Abstract

Both nitric oxide (NO) and reactive oxygen species (ROS) are versatile molecules that mediate a variety of cellular responses in plants. In this chapter, methods for imaging NO and ROS using laser scanning confocal microscopy (LSCM) are presented. Arabidopsis roots, dyed with DAF-FM or H2DCF, are observed using the Leica TCS-SP2 LSCM. NO or ROS production are imaged and their kinetic changes monitored with the laser excitation and emission wavelengths at 488 nm and between 500 and 530 nm, respectively. In addition, Leica software is employed to visualize and calculate the fluorescence intensity data.