Fluorochromes and fluorescence

Abstract

To facilitate flow cytometry, there is a need for cells to be labeled with fluorochromes. However, because of the spectral properties of fluorochromes, the fluorescence associated with one fluorochrome may be detected by more than one detector, producing spectral overlap which requires compensation. However, the number of useful fluorochromes has increased steadily over the years, and more recently there have been significant advances in techniques for compensating for spectral overlap between several fluorescent signals, so it is likely that the number of fluorochromes used in analysis will increase further. The basic principles of how light and matter interact, the nature of fluorescence, and the properties of various fluorochromes that are used for different purposes are described in this chapter. Some examples of the many parameters that can be measured using fluorescence and the fluorescent probes that can be used are also given.