Employment of a mereyanine dye for the detection of malignant leukocytic cells

• US 4,424,201 A
• Filed: 11/28/1978
• Issued: 01/03/1984
• Est. Priority Date: 11/28/1978
• Status: Expired due to Term

• Current Assignee: Rockefeller University
• Original Assignee: Rockefeller University
• Inventors: Valinsky, Jay E., Easton, Thomas G., Reich, Edward
• Primary Examiner(s): Fagelson, Anna P.
• Application Number:
  US05/964,359
• Time in Patent Office: 1,862 Days
• Field of Search: 424/1, 424/1.5, 424/9, 424/3, 424/7, 435/34, 435/35, 436/64, 436/800, 436/804, 436/813
• US Class Current: 435/34
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/804 Radioisotope, e.g. radioimmunoassay
• CPC Class Codes: Y10S 436/813 Cancer

Abstract

A method for the detection of malignant leukocytic cells which comprises:

contacting a fluid containing malignant leukocytic cells with a merocyanine dye of the formula: ##STR1## wherein n=1-5; X=0 or S; R₁ and R₂ are the same or different C₂ -C₅ alkyl groups:

wherein said dye is incorporated into said cells; separating the cells from the incubation mixture; and detecting said dyed cells. Fluorescence or radioactivity detection methods are preferred detecting means.

Claims

1. A method for the detection of malignant leukocytic cells which comprises contacting a fluid solution containing malignant leukocytic cells with a merocyanine dye of the formula ##STR5## wherein n=1-5;

X=0 or S and R₁ and R₂ are the same or different C₂ -C₅ alkyl groups;

wherein said dye is incorporated into said cells;

separating the cells from the incubation mixture and then detecting said dyed cells.

2. The method of claim 1 wherein n=3;

X=S;

R₁ =R₂ =--C₄ H₉.

3. The method of claim 1 wherein said detection is by fluorescence.

4. The method of claim 3 wherein said detection of fluorescence is carried out by darkfield-fluorescence microscopy.

5. The method of claim 3 wherein said detection of fluorescence is carried out by flow microfluorimetric methods.

6. The method of claim 3 wherein said fluorescence is produced by excitation radiation of a wavelength in the range 400-700 nm.

7. The method of claim 1 wherein said contact is effected in a saline solution.

8. The method of claim 7 wherein said saline solution contains autologous plasma.

9. The method of claim 1 wherein the concentration of said cells in said fluid is in the range 10⁵ to 5×

10⁷ cells/ml.

10. The method of claim 1 wherein the concentration of said dye in said solution is in the range 0.1 to 200 μ

g/ml.

11. The method of claim 1 wherein the temperature of contacting is 0°

to 40°

C.

12. The method of claim 1 wherein said merocyanine dye is radioactive.

13. The method of claim 12 wherein said merocyanine dye is that wherein X=³⁵ S.

14. The method of claim 11 wherein said detection is done by determining radioactivity uptake of dye by said leukocytic cells present in said fluid solution.

15. The method of claim 11 wherein the specific activity of said radioactive dye in said solution is 0.1 to 100 Ci/mole.

16. The method of claim 1 wherein said contacting is carried out in the presence of ambient illumination.

17. The method of claim 1, wherein the incubation of said merocyanine dye with said fluid solution is carried out in the presence of trypan blue dye.

18. The method of claim 1 wherein said malignant leukocytic cells are leukemic cells which are selected from the group consisting of chronic and acute leukemias of the lymphocytic, myelocytic, myelocyticmonocytic, monocytic, nonlymphoblastic, hystocytic and lymphoma types.

19. A method of detecting solid tumors which comprises contacting a fluid solution containing leukocytic cells derived from the blood of patients having, suspected of having or having had a solid malignant tumor, with a merocyanine dye of the formula ##STR6## wherein n=1-5;

X=0 or S, and R₁ and R₂ are the same or different C₂ -C₅ alkyl groups;

wherein said dye is incorporated into said cells;

separating the cells from the incubation mixture and detecting said dyed cells.