7 - Fluorescence Microscopy in Three Dimensions: Microtomoscopy

Abstract

This chapter discusses the microtomoscopy and its application. In combination with specific labeling of cellular components, confocal fluorescence microscopy is a new method for quantifying molecular biological processes in live cells. The major reason for this is the three-dimensional imaging capability of such a microscope. This means that the three-dimensional distribution of any fluorophore can be measured directly, without the need for cumbersome and noise-producing computational methods. It also means that the microscope produces images that do not contain any out-of-focus information and is called optical sections. Currently this kind of microscopy is moving from the development state, where only groups interested in the development of new methods in microscopy have such instruments, to the state where any biologist can operate it. Apart from the special optics, an image-processing computer is an integral part of the microscope, which opens a large number of applications for analytical microscopy. Various commercial systems are now appearing on the market, both object-scanning systems and beam-scanning systems. All of these systems are suitable for biological applications and bring the possibilities of confocal microscopy within the realm of any cell biologist.