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FITC is a yellow crystal with a maximum absorption wavelength of 490-495nm and a maximum emission wavelength of 520-530nm. It can present bright yellow green fluorescence. FITC can be preserved for many years in a cold dark and dry place, and is currently the most widely used fluorescein. Its main advantage is that the human eye is more sensitive to yellow green, usually the green fluorescence in the section specimen is less than red.

FITC molecule contains isothiocyano. When FITC reacts with antibody protein in alkaline solution, it is mainly the R amino group of lysine on protein combines with thiocarboside of fluorescein to form FITC protein conjugate, i.e. fluorescent antibody or fluorescent conjugate.

Yellow green fluorescence is produced under the excitation of ultraviolet or blue violet light of fluorescent lamp source. The corresponding antigen can be qualitatively, quantitatively detected by observing under fluorescence microscope or analyzing by flow cytometry.