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Live cell imaging is an essential methodology for studying the structure, dynamics, and functions of cells in a living plant under normal or stressed growth conditions. Arabidopsis thaliana is perfectly amenable to various live microscopy techniques. In this chapter, we provide guidelines to design live-imaging experiments. We discuss specifically the respective advantage of each microscopy technique, the choice of reporter, and the preparation of the sample. Detailed protocols for imaging of shoot and roots are provided.

Identification and characterization of differentially expressed genes is required in many fields of modern biology and medicine. Therefore, efficient methods have been developed for identification and isolation of differentially expressed genes using molecular tools. Differential display of mRNA by PCR (DD-PCR) is a new and powerful procedure for quantitative detection of differentially expressed genes (Liang and Pardee 1992; McClelland et al. 1995). Advantages over alternative procedures include: (1) simultaneous display of all differences in gene expression between different cell fractions; (2) simultaneous detection of both up-regulation and down-regulation of genes; (3) requirement of only small amount of sample tissue; and (4) drastically reduced time of analysis. The DD-PCR procedure is based on reverse transcription of mRNA from different cell fractions using an oligo-dT-NN anchor primer followed by PCR amplification of cDNA using a set of short arbitrary primers and radiolabeled dNTP. After electrophoretic separation of the resulting fragments on sequencing gels, differential gene expression is visualized by autoradiography. Differentially expressed cDNA species can be recovered from the gel using the autoradiogram for band localization.