ABSTRACT
The development of the laser scanning confocal microscopy revolutionized the field of life sciences, since this technology allows the generation of sharper images with significant lower background. This chapter focuses on the description of two common applications of confocal microscopy within the life science field such as colocalization and fluorescence recovery after photobleaching (FRAP) and a summary of commonly used fluorophores. The observation of biological samples by confocal microscopy should, ideally, be carried out in living specimens. After adequate sample preparation appropriate image acquisition begins with a clear idea of the purpose of the image and the purpose of showing the most representative images of the sample. The permeabilization of the sample has the objective of granting access of labeling agents to intracellular molecules or components. The use of fluorescently conjugated antibodies is a widespread practice for the labeling of specific macromolecules to perform immunofluorescent assays.
