Methods for qualitative and quantitative analysis of cells and related optical bio-disc systems

• US 20030129665A1
• Filed: 08/29/2002
• Published: 07/10/2003
• Est. Priority Date: 08/30/2001
• Status: Abandoned Application

• Current Assignee: Vindur Technologies, Inc.
• Original Assignee: Vindur Technologies, Inc.
• Inventors: Selvan, Gowri Pyapali, Gordon, John Francis, Urcia, Joseph Roby Iringan, Brazil, Karen Jean
• Application Number:
  US10/230,959
• Publication Number:
  US 20030129665A1
• Time in Patent Office: Days
• US Class Current: 435/7.2
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 35/00069 whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk

Abstract

A clinical diagnostic assay is performed on an optical bio disc and is read with a disc drive. Methods and apparatus for detecting and quantifying specific blood cell analytes in biological samples using optical bio disc is disclosed. The method for determining the quantity of a specific type of blood cell in a biological sample includes binding an antibody to a capture zone on the disc, providing a sample to the capture zone, remove portions of the sample that are not bound in the capture zone, and counting bound cells. Also described is method and apparatus for performing a cluster designation count with an optical disc and disc drive and method for making an optical assay disc for performing such cluster designation count.

Claims

1. A system for detecting blood cell analytes in biological samples, said system comprising:

an optical disc having a substrate;

a cap parallel to said substrate;

a chamber between said substrate and cap, said chamber including capture zones;

a capture layer associated with said substrate at the capture zones, such that a first capture zone has first cell capture agents and a second capture zone has a second cell capture agents;

a light source for directing light to the disc at the capture zones;

a detector for detecting light reflected from or transmitted through the disc at the capture zones and providing a signal; and

a processor for using the signal to count items in the sample bound to the capture agents.

2. A method for detecting blood cell analytes in biological samples employing an optical disc and disc drive, said method comprising the steps of:

providing a sample of cells on a disc surface in a chamber in of the disc, the chamber including at least one capture zone with a capture agent;

loading the disc into an optical reader;

rotating the optical disc;

directing an incident beam of electromagnetic radiation toward the capture zone;

detecting a beam of electromagnetic radiation formed after interacting with the disc at the capture zone;

converting the detected beam into an output signal; and

analyzing the output signal to extract therefrom information relating to the number of cells captured at the capture zone.

3. The method according to claim 2 wherein the chamber with the disc surface supporting the sample is internal to the disc and is bounded on opposite sides by a substrate and cap.

4. The method according to claim 2 wherein the optical disc is constructed with a reflective layer such that light directed to the capture zone and not striking a cell is reflected.

5. The method according to claim 2 wherein the optical disc is constructed such that light directed to the capture zone and not striking a cell is transmitted through the optical disc, the disc being between the light source and the detector.

6. The method according to claim 2 wherein the disc surface is coated with a first group of cell capture agents.

7. The method according to claim 6 wherein the cell capture agents define a discrete capture zone.

8. The method according to claim 7 wherein a second group of cell capture agents define a second discrete capture zones in a predetermined pattern.

9. The method according to claim 8 wherein the first and second capture zones are in one chamber.

10. The method according to claim 6 wherein the cell capture agents are for binding with cell surface antigens.

11. The method according to claim 10 wherein the cell surface antigen is selected from the CD family of antigens.

12. A method of performing a cluster designation count with an optical disc and disc drive, said method comprising the steps of:

providing a blood sample in a first tube having a separation gradient;

rotating the first tube at a time and speed sufficient to separate the blood sample into layers;

resuspending an MNC layer containing T-cells to thereby form an MNC suspension;

providing a sample of the MNC suspension on an optical disc surface including at least one capture zone with at least one capture agent;

loading the optical disc into an optical reader;

rotating the optical disc;

directing an incident beam of electromagnetic radiation toward said at least one capture zone;

detecting a beam of electromagnetic radiation formed after interacting with the disc at the capture zone;

converting the detected beam into an output signal; and

analyzing the output signal to extract therefrom information relating to the number of cells captured at the capture zone.

13. The method according to claim 12 wherein the separation gradient is provided by a matrix that forms a density gradient upon centrifugation.

14. The method according to claim 12 wherein the first tube further contains an anticoagulant.

15. The method according to claim 14 wherein the anticoagulant is EDTA.

16. The method according to claim 14 wherein the anticoagulant is acid citrate dextran.

17. The method according to claim 14 wherein the anticoagulant is heparin.

18. The method according to claim 12 wherein the surface is internal to the disc and is bounded on opposite sides by a substrate and a cap.

19. The method according to claim 12 wherein the optical disc is constructed with a reflective layer such that light directed to the capture zone and not striking a cell is reflected.

20. The method according to claim 12 wherein the optical disc is constructed such that light directed to the capture zone and not striking a cell is transmitted through the optical disc, the disc being between the light source and the detector.

21. The method according to claim 12 wherein the disc surface is coated with a first group of capture agents.

22. The method according to claim 21 wherein the capture agents are antibodies.

23. The method according to claim 22 wherein the antibodies are selected from the group including monoclonal, polyclonal, and recombinantly created antibodies.

24. The method according to claim 21 wherein the capture agents are immobilized on the disc surface by a cross-linking system.

25. The method according to claim 20 wherein the capture agents are immobilized directly on the disc surface.

26. The method according to claim 20 wherein the capture agents define one or more discrete capture zones.

27. The method according to claim 26 wherein the one or more capture zones are located within one or more chambers within the optical disc.

28. The method according to claim 20 wherein the capture agents have a selective affinity for cell surface antigens.

29. The method according to claim 28 wherein the cell surface antigens are selected from the CD family of antigens.

30. The method according to claim 29 wherein the cell surface antigens are selected from the group consisting of CD3, CD4, CD8, and CD45.

31. The method according to claim 12 wherein the blood sample comprises normal blood cells, leukemic blood cells, or lymphomic blood cells.