Patent ID: 8367358

Claim:
A reagent for differentiating and counting leukocytes, said reagent comprises: (1) cationic cyanine compounds selected from those having the following general formulae I and II; wherein n is 1, 2 or 3; X is C(CH 3 ) 2 , O, S or Se; R 1 and R 2 are each independently selected from H, C 1-18 alkyl, —C 1-6 alkyl-OR 5 or a halogen; R 3 is H, C 1-18 alkyl, OR 5 , —C 1-6 alkyl-OR 5 , COOR 5 , NO 2 , CN or a halogen; R 4 is C 1-18 alkyl, —C 1-6 alkyl-OR 5 , benzyl or a halogen, wherein said benzyl is optionally substituted with a substituent selected from a halogen, hydroxyl, sulfhydryl, cyano, nitro, alkyl, aryl, alkyloxy, haloalkyl, amino, alkylamino, amido or carboxyl; R 5 is H or C 1-18 alkyl; and Y − is an anion; or wherein n is 1, 2 or 3; X is C(CH 3 ) 2 , O, S or Se; R 1 ′ and R 2 ′ are each independently selected from H, OH, C 1-18 alkyl, C 1-6 alkylOR 5 ′, C 1-18 alkylsulfonyl, phenyl or a halogen; R 3 ′ and R 4 ′ are each independently selected from C 1-18 alkylCOOR 6 ′, C 1-18 alkylOR 6 ′ or benzyl, wherein said benzyl is optionally substituted with a substituent selected from a halogen, hydroxyl, sulfhydryl, cyano, nitro, alkyl, aryl, alkyloxy, haloalkyl, amino, alkylamino, amido or carboxyl, provided that R 3 ′ and R 4 ′ are not simultaneously benzyl, and R 4 ′ is not C 1-18 alkylOR 6 ′ when R 3 ′ is benzyl; R 5 ′ is C 1-18 alkyl or H; R 6 ′ is C 1-18 alkyl, H or phenyl, wherein said phenyl is optionally substituted with a substituent selected from a halogen, hydroxyl, sulfhydryl, cyano, nitro, alkyl, aryl, alkyloxy, haloalkyl, amino, alkylamino, amido or carboxyl; and Y − is an anion; (2) cationic surfactants that are quinolinium salt-type cationic surfactants having the following general formulae III and/or IV: wherein R 1 and R 2 are each independently selected from C 6-18 alkyl and C 6-18 haloalkyl; R 3 to R 16 are each independently selected from H, OH, C 1-4 alkyl, C 1-4 alkyloxy and sulphonyl; and Z − is a halogen ion; (3) at least one nonionic surfactant; and (4) at least one anionic compound selected from those having one or more carboxyl or sulphonyl groups.