ABSTRACT
This chapter looks at the possibilities and applications of real-time confocal fluorescence microscopy for basic research and clinical applications in animal and human tissues in vivo. It provides a brief introducion to the main concepts of confocal imaging and lists the original references that contain the detailed theory. A confocal microscope is made up of a small, bright source of light that illuminates a small three-dimensional spot within the object. Real-time high-resolution confocal fluorescence imaging in tissue in vivo is challenging. At fluorophore concentration that is low enough to be nontoxic to the tissue, the very small illuminated confocal spot may not contain enough fluorophore molecules to produce a strong fluorescence signal. Photobleaching occurs when the triplet state in the fluorophore reacts with molecular oxygen, forming a nonfluorescing molecule. Longer detector integration times can be obtained at the expense of resolution with a line-scanning confocal microscope, in which an illumination line is scanned.
