53 - Tissue-Print Hybridization on Membrane for Localization of mRNA in Plant Tissue

Abstract

Publisher Summary This chapter discusses the tissue-print hybridization on membrane for the localization of messenger ribonucleic acid (mRNA) in a plant tissue. When a freshly cut section of the plant tissue is pressed on a nitrocellulose or nylon membrane, the soluble contents of the cut cells are transferred to the membrane. The cell contents on the membrane make a latent print that can then be visualized with appropriate probes. The rigid cell walls leave a physical print that makes the anatomy of the tissue visible without any further treatment. The tissue print technique allows rapid and convenient tissue-level localization of proteins, enzymes, mRNAs, viral DNA, and metabolites. The advantage of the tissue-print technique is that the fresh tissue can be cut and the freshly cut surface can be pressed on the membrane immediately without any treatment. The cellular mRNAs are immobilized on the membrane and are then detected by specific probes. The specific cellular mRNAs show little lateral diffusion during or after printing on the membrane. The precise localization of the mRNAs can then be determined by comparison with the anatomy of corresponding stained sections or by comparison with the physical print on the membrane. Another advantage of the tissue print technique is that large organs or whole seedlings can be printed on the same membrane and the spatial distribution of specific mRNAs be analyzed.