Method for labeling and detecting materials employing arylsulfonate cyanine dyes

• US 5,486,616 A
• Filed: 11/29/1993
• Issued: 01/23/1996
• Est. Priority Date: 04/18/1986
• Status: Expired due to Term

• Current Assignee: Carnegie Mellon University
• Original Assignee: Carnegie Mellon University
• Inventors: Ernst, Lauren A., Mujumdar, Ratnakar B., Waggoner, Alan S.
• Primary Examiner(s): Gerstl, Robert
• Application Number:
  US08/158,952
• Time in Patent Office: 785 Days
• Field of Search: 548/217, 548/219, 548/150, 548/159
• US Class Current: 548/217
• CPC Class Codes: C09B 23/0008 substituted on the polymethine chain
• CPC Class Codes: C09B 23/02 the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
• CPC Class Codes: C09B 23/10 The polymethine chain containing an even number of >CH- groups styryl dyes C09B23/14, C09B23/14 takes precedence
• CPC Class Codes: C09B 23/145 the ethylene chain carrying an heterocyclic residue, e.g. heterocycle-CH=CH-C6H5
• CPC Class Codes: C09B 62/0025 Specific dyes not provided for in groups C09B62/004 - C09B62/018
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

A method for detecting a component of an aqueous liquid comprising adding to the liquid a luminescent dye selected from the group consisting of cyanine, merocyanine and styryl dyes containing at least one sulfonic acid or sulfonate group attached to an aromatic nucleus and reacting the dye with the component. The labeled component is then detected by an optical detection method.

Claims

1. A water soluble luminescent dye consisting of:

a cyanine having the structure ##STR11## wherein;

the dotted lines each represent carbon atoms necessary for the formation of one ring to three fused rings having 5 to 6 atoms in each ring and said R₃, R₄, R₈ and R₉ groups are attached to said rings;

at least one of X and Y is oxygen and the other is selected from the group consisting of O, S and CH₃ --C--CH₃ ;

m is an integer selected from the grouping consisting of 1, 2 and 3;

at least one of said R₁, R₂, R₃, R₄ and R₇ groups reacts with amino, hydroxy or sulfhydryl nucleophiles and is a reactive moiety selected from the group consisting of ##STR12## wherein Q is Br or Cl, and n=0, 1, 2, 3, 4, 5, 6, 7 or 8 for R₃, R₄ and R₇, and n=1, 2, 3, 4, 5, 6, 7 or 8 for R₁ and R₂ ;

and when any one of said R₃, R₄, and R₇ groups is not selected from one said reactive moiety, said remaining R₃, R₄, and R₇ are hydrogen or E--F, a polar group wherein E is a spacer group having the structure --(CH₂)_n -- and n of said spacer group=0, 1, 2, 3, or 4, and F is hydroxy, sulfonate, sulfate, carboxylate, or a lower alkyl substituted amino;

and when any one of said R₁ and R₂ groups is not selected from one said reactive moiety, said remaining R₁ or R₂ are a lower alkyl or said E--F and n of said spacer group=1, 2, 3, or 4;

at least one said R₈ and R₉ groups is selected from the group consisting of a sulfonic acid and sulfonate moiety attached directly to one of said one ring to three fused rings for conferring luminescence to said dye, wherein said ring or rings to which said sulfonic acid or sulfonate moiety is attached is an aryl ring;

and when one of said R₈ or R₉ group is not a sulfonic acid or sulfonate, said remaining R₈ or R₉ group is a hydrogen.

2. The luminescent dye of claim 1 which in water has a molar extinction coefficient of at least 50,000 liters per mole centimeter, a quantum yield of at least 2 percent and absorbs and emits light in the 400 to 900 nm spectral range.

3. The luminescent dye of claim 1 which in water has a molar extinction coefficient of at least 100,000 liters per mole centimeter, a quantum yield of at least 10 percent and absorbs and emits light in the 600 to 900 nm spectral range.

4. The luminescent dye of claim 1 wherein said cyanine has the structure:

##STR13##

5. The luminescent dye recited in claim 1 wherein the cyanine has the structure:

##STR14## wherein R₁₀ is selected from the group consisting of said reactive moiety, hydrogen and a sulfonic acid or sulfonate moiety.

6. A water soluble luminescent dye consisting of a cyanine having the structure ##STR15## wherein:

at least one of X and Y is oxygen and the other is selected from the group consisting of O, S and CH₃ --C--CH₃ ;

m is an integer selected from the group consisting of 1, 2 and 3;

at least one of said R₁, R₂, R₃, R₄, R₇, R₁₀ and R₁₁ groups reacts with amino, hydroxy or sulfhydryl nucleophiles and is a reactive moiety selected from the group consisting of;

##STR16## wherein Q is Br or Cl, and n=0, 1, 2, 3, 4, 5, 6, 7 or 8 for R₃, R₄, R₇, R₁₀ and R₁₁, and n=1, 2, 3, 4, 5, 6, 7 or 8 for R₁ and R₂ ;

and when any one of said R₃, R₄, and R₇ groups is not selected from one said reactive moiety, said remaining R₃, R₄, and R₇ are hydrogen or E--F, a polar group wherein E is a spacer group having the structure --(CH₂)_n -- and n=0, 1, 2, 3, or 4, and F is hydroxy, sulfonate, sulfate, carboxylate, or lower alkyl substituted amino;

and when any one of said R₁ and R₂ groups is not selected from one said reactive group, said remaining R₁ and R₂ are lower alkyl or said E--F and n=1, 2, 3, or 4;

said R₈ and R₉, and said R₁₀ and R₁₁ groups when said R₁₀ and R₁₁ groups are not said reactive moiety, are selected from the group consisting of hydrogen and a sulfonic acid or sulfonate moiety, wherein at least one of said R₈, R₉, R₁₀ and R₁₁ groups is one of said sulfonic acid or said sulfonate moiety attached directly to an aryl ring of said dye structure for conferring luminescence to said dye.

7. A water soluble luminescent dye consisting of:

##STR17## wherein at least one of X and Y is oxygen and the other is selected from the group consisting of O, S and CH₃ --C--CH₃ ;

m is an integer selected from the group consisting of 1, 2 and 3;

at least one of said R₁ and R₂ is ##STR18## and the other is selected from the group consisting of ##STR19## where n=1, 2, 3, 4, 5, 6, 7 or 8 and k=2, 3, 4 or 5; and

at least one of said R₈ and R₉ groups is a sulfonic acid or sulfonate moiety attached directly to an aryl ring of said dye for conferring luminescence to said dye;

and when one of said R₈ and R₉ group is not said sulfonic acid or sulfonate moiety, said remaining R₈ or R₉ group is a hydrogen.

8. A water soluble luminescent dye consisting of:

##STR20## wherein at least one of X and Y is oxygen and the other is selected from the group consisting of O, S and CH₃ --C--CH₃ ;

m is an integer selected from the group consisting of 1, 2 and 3;

at least one of said R₁ and R₂ is ##STR21## and the other is selected from the group consisting of ##STR22## where n=1, 2, 3, 4, 5, 6, 7, or 8 and k=2, 3, 4, or 5;

at least two of said R₈, R₉, R₁₀ and R₁₁ groups are a sulfonic acid or sulfonate moiety attached directly to an aryl ring of said dye for conferring luminescence to said dye;

and when any one of said R₈, R₉, R₁₀ and R₁₁ groups is not said sulfonic acid or sulfonate moiety, it is a hydrogen.

9. A water soluble luminescent dye consisting of:

##STR23## wherein at least one of X and Y is oxygen and the other is selected from the group consisting of O, S and CH₃ --C--CH₃ ;

m is an integer selected from the group consisting of 1, 2, and 3;

at least one of said R₁ and R₂ is ##STR24## and the other is selected from the group consisting of ##STR25## where n=1, 2, 3, 4, 5, 6, 7 or 8 and k=2, 3, 4 or 5;

at least two of said R₈, R₉ and R₁₀ groups are a sulfonic acid or sulfonate moiety attached directly to an aryl ring of said dye for conferring luminescence to said dye;

and when any one of said R₈, R₉ or R₁₀ is not said sulfonic acid or sulfonate moiety, it is a hydrogen.