Cyanine-labeling reagents: sulfobenzindocyanine succinimidyl esters.

A synthetic method for shifting the absorption and emission wavelengths of cyanine dye labels by 15-30 nm to the red has been developed. This step significantly increases the potential for preparing fluorescent probes for multiparameter analysis in cytometry and diagnostics. The new sulfobenzindocyanine dyes contain succinimidyl esters as reactive groups and can be readily conjugated to antibodies, avidin, modified DNA, and other amino group-containing materials. The labeling reagents are water soluble, and their fluorescence is not sensitive to pH. One of the reagents, Cy3.205, can be optimally excited with the 568 nm Kr laser line and is useful for confocal microscopy and flow cytometry. Another dye, Cy5.205, can be excited with the 633 He-Ne or 647 nm Kr laser lines available with many flow cytometers and laser-scanning microscopes. New laser diodes emitting near 660-690 nm should also be excellent excitation sources for Cy5.205.