A fluorescent dye comprising a xanthene-derived fluorophore having the formula (I) wherein R1, R2, R3, R4, R5 and R6 are independently selected from H, alkyl, alkoxy, alcohol, ether, alkenyl, alkenoxy, aryl, alkaryl, aralkyl and amido, except that R1, R4 and/or R5 is not H when bonded to Y, Y1 or Y2, respectively; X is either O− or S−; and at least one of Y, Y1, Y2, Y3, Y4 and Y5 is a group for covalently bonding the dye, optionally through. the use of a coupling agent, to a target molecule, and is otherwise H. The dye may be covalently attached to a target molecule to form a complex, and the dye and/or complex finds use in cell analysis techniques, particularly pH measurement and analysis of kinetics of migration.