Chapter 15 Confocal Microscopy of the Shoot Apex

Abstract

This chapter focuses on confocal laser scanning microscopy (CLSM). In CLSM, a sample is treated with a fluorescent dye, and a laser beam is focused on a point in the sample, causing the emission of fluorescent light. The use of CLSM in the study of plant development has many advantages over conventional histological techniques. Because CLSM data are stored digitally, the data can not only be reconstructed in three dimensions, but also animated or displayed as stereo images, to aid in visual interpretation. Once the sample is reconstructed on a computer, virtual sections can be made from any orientation. Specimen preparation for CLSM is also much less labor intensive compared to serial sectioning, and avoids problems such as recovering and mounting individual sections, a time-consuming and error-prone task. The integrity of the sample is preserved in CLSM, allowing rescanning using different parameters. The chapter describes the method of examining plant tissue using CLSM, particularly in studies of the structure, cell number, and cell pattern of shoot apical meristems and floral meristems. The development of this technique is timely because of the recent explosion of interest in developmental mutants that affect apical and floral meristem structure.