Overview of Fluorophores

Since its inception in the 1940s, the technique of immunofluorescence has provided a sensitive, high-resolution method for analyzing the binding of antibody to antigen. Fluorescent molecules, termed fluorophores or fluorochromes, can be conjugated directly to antibodies by covalent linkage, or coupled indirectly to antibodies via conjugation to proteins A and G or through an avidin-biotin bridge (see Chapter 6 ,Chapter 7 ,Chapter 21 , and Chapter 22 ). The basic features of immunofluorescence are straightforward, but a working knowledge of the commonly used fluorophores is of value in obtaining maximum performance from immunofluorescence microscopy and flow cytometry.