Detection of reticulocytes, RNA or DNA

• US 4,883,867 A
• Filed: 08/03/1987
• Issued: 11/28/1989
• Est. Priority Date: 11/01/1985
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Lee, Linda G., Chen, Chia-Huei
• Primary Examiner(s): Sneed, H. M. S.
• Application Number:
  US07/081,097
• Time in Patent Office: 848 Days
• Field of Search: 250/461, 546/152, 546/166, 546/176, 548/146, 435/4, 435/6, 436/63, 436/72, 436/800, 436/10, 436/94, 436/172, 424/3, 424/7.1, 424/9, 356/39, 536/28
• US Class Current: 536/25.4
• CPC Class Codes: G01N 33/80 involving blood groups or blood types or red blood cells white blood cells G01N33/56972
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

Reticulocytes, RNA or DNA are stained with a dye for detection in a flow cytometer. The dye has the formula: ##STR1## Wherein: X=O,S, Se, N -alkyl (having 1-6 carbons) or C (CH₃)_n ; R₁ =alkyl having from 1-6 carbons;

R₂ =alkyl having from 1-6 carbons;

R₃ =fused benzene, alkyl (having 1-6 carbons), methoxy or is absent;

R₄ =alkyl having 1-6 carbons, methoxy or is absent; and

n=zero or an integer from 1-6.

Claims

1. A compound composed of reticulocytes bound to a dye having the formula:

##STR7## Wherein;

X═

O,S, Se or C (CH₃)₂ ;

R₁ =alkyl having from 1-6 carbons;

R₂ =alkyl having from 1-6 carbons;

R₃ =fused benzene, alkyl (having 1-6 carbons), methoxy or hydrogen;

R₄ =alkyl having 1-6 carbons, methoxy or hydrogen; and

n=zero or an integer from 1-6.

2. A compound of claim 1 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ =hydrogen, R₄ =hydrogen, X═

S and n=1.

3. A compound in accordance with claim 1 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ ═

fused benzene, R₄ ═

hydrogen, X═

S and n=0.

4. A compound in accordance with claim 1 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ ═

hydrogen, R₄ ═

hydrogen, X=0 and n=zero.

5. A compound composed of RNA bound to a dye having the formula:

##STR8## Wherein;

X═

O,S, Se or C (CH₃)₂ ;

R₁ =alkyl having from 1-6 carbons;

R₂ =alkyl having from 1-6 carbons;

R₃ =fused benzene, alkyl (having 1-6 carbons), methoxy or is hydrogen;

R₄ =alkyl having 1-6 carbons, methoxy or hydrogen; and

n=zero or an integer from 1-6.

6. A compound in accordance with claim 5 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ ═

hydrogen, R₄ ═

hydrogen, X═

S, and n=1.

7. A compound in accordance with claim 5 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ ═

fused benzene, R₄ ═

hydrogen, X═

S and n=0.

8. A compound in accordance with claim 5 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ ═

hydrogen, R₄ ═

hydrogen, X=0 and n=zero.

9. A compound composed of DNA bound to a dye having the formula:

##STR9## wherein;

X═

O,S, Se or C (CH₃)₂ ;

R₁ =alkyl having from 1-6 carbons;

R₂ =alkyl having from 1-6 carbons;

R₃ =fused benzene, alkyl (having 1-6 carbons), methoxy or is hydrogen;

R₄ =alkyl having 1-6 carbons, methoxy or hydrogen; and

n=zero or an integer from 1-6.

10. A compound in accordance with claim 9 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ ═

hydrogen, R₄ ═

hydrogen, X═

S and n=1.

11. A compound in accordance with claim 9 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ =fused benzene, R₄ =hydrogen, X═

S and n=0.

12. A compound in accordance with claim 9 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ =hydrogen, R₄ =hydrogen, X=0 and n=zero.

13. A compound in accordance with claim 9 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃ =hydrogen, R₄ =hydrogen, X═

S and n=1.

14. A compound in accordance with claim 9 wherein R₁ ═

CH₂ C₂ H₅, R₂ ═

CH₂ CH₃, R₃ =hydrogen, R₄ =hydrogen, X═

S and n=0.

15. A compound in accordance with claim 9 wherein R₁ ═

CH₃, R₂ ═

CH₃, R₃,═

OCH₃, R₄ ═

OCH₃, X═

S and n=0.