Cy5 alkyne, with low aqueous solubility is a dye ready for the use in Click Chemistry reaction. Various molecules via Click Chemistry reaction can be attached by this deeply colored, and photostable Cy5 fluorophore. Most derivatives of non-sulfonated cyanines have low aqueous solubility except for hydrochlorides of hydrazides and amines. For biomolecule labeling, using of organic co-solvent to dissolve this molecular is necessary for efficient reaction. First, Cyanine dye should be dissolved in organic solvent and then added to a solution of biomolecule in appropriate aqueous buffer. Various substrates such as biomolecules, polymers, and solid surfaces which bearing azides can be used for the labeling.