Capture layer assemblies for cellular assays including related optical analysis discs and methods

• US 20030143637A1
• Filed: 08/30/2002
• Published: 07/31/2003
• Est. Priority Date: 08/31/2001
• Status: Abandoned Application

• Current Assignee: Nagaoka & Co., Ltd.
• Original Assignee: Nagaoka & Co., Ltd.
• Inventors: Brazil, Karen Jean, Selvan, Gowri Pyapali, Gordon, John Francis, Urcia, Joseph Roby Iringan
• Application Number:
  US10/233,322
• Publication Number:
  US 20030143637A1
• Time in Patent Office: Days
• US Class Current: 435/7.9
• 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 15/1475 using image analysis for extracting features of the particle
• CPC Class Codes: G01N 2015/1486 Counting the particles
• CPC Class Codes: G01N 2021/0325 Cells for testing reactions, e.g. containing reagents
• CPC Class Codes: G01N 2021/1742 either absorption or reflection
• CPC Class Codes: G01N 2021/551 Retroreflectance
• CPC Class Codes: G01N 2021/5915 Processing scan data in densitometry
• CPC Class Codes: G01N 21/07 Centrifugal type cuvettes G01N21/09 takes precedence
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/55 Specular reflectivity
• CPC Class Codes: G01N 21/59 Transmissivity G01N21/25 takes precedence
• CPC Class Codes: G01N 21/5911 of the scanning type scanning per se G02B
• CPC Class Codes: G01N 2201/104 Mechano-optical scan, i.e. object and beam moving
• 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. More specifically this invention relates to capture layer assemblies for cellular assays using optical bio-discs. The current invention includes methods for determining the quality and quantity of a specific type of cell in a biological sample. These methods includes binding capture agents 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. The current invention also includes methods and apparatus for performing a cluster designation marker assay using an optical disc and disc drive and method for making optical analysis discs for performing such cluster designation assays.

Claims

1. A method of conducting an assay with an optical disc and disc drive, the method comprising:

providing a sample of cells on a disc surface in a chamber in a 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 to 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.

2. A method of conducting an assay with an optical disc and disc drive, the method comprising:

providing a sample of cells on a disc surface in a chamber in a disc, the chamber including capture zone with a capture layer assembly;

rotating the optical disc;

directing an incident beam of electromagnetic radiation to 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 capture layer assembly comprises a first layer;

a second layer;

a secondary capture antibody and a primary capture antibody.

4. The method according to claim 3 wherein the first layer is made of polystyrene.

5. The method according to claim 3 wherein the first layer is made of streptavidin.

6. The method according to claim 3 wherein the second layer is streptavidin.

7. The method according to claim 3 wherein the secondary capture antibody is B-antimouse IgG.

8. The method according to claim 7 wherein the secondary capture antibody binds with a cross linking agent.

9. The method according to claim 8 wherein the cross-linking agent is aldehyde activated dextran.

10. The method according to claim 3 wherein the primary capture antibody is selected from the group comprising anti-CD3 antibody, biotinylated-anti-CD3 antibody, anti-CD4 antibody, biotinylated-anti-CD4 antibody, anti-CD8 antibody, biotinylated-anti-CD8 antibody, anti-CD45 antibody, and biotinylated-anti-CD45 antibody.

11. An optical disc and drive system for receiving a sample, the system comprising:

a disc including;

a substrate;

a cap parallel to the substrate, a chamber defined therebetween and including capture zones;

a capture layer over the 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 molecules.

12. A method of making an optical assay disc for performing a cluster designation count, said method comprising:

providing a cross-linker in a tube;

adding a capture agent to said tube;

allowing said cross-linker and said capture agent to combine, thereby forming a complex;

providing a substrate;

coating said substrate with an active layer;

depositing said complex onto said active layer; and

attaching a cap portion to said active layer using an adhesive member.

13. A method of conducting an assay with an optical disc and drive system, said method comprising:

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

rotating the optical disc;

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

detecting the 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.

14. The method according to claim 13 wherein said disc surface is provided by a metal layer.

15. The method according to claim 13 wherein said disc surface is provided by a polycarbonate substrate.

16. The method according to claim 14 wherein said capture layer assembly is associated with said metal layer.

17. The method according to claim 16 wherein said capture layer assembly includes an active layer.

18. The method according to claim 17 wherein said active layer is formed from a material selected from the group comprising polystyrene, glass, nylon, polycarbonate, streptavidin, neutravidin, avidin, biotin, and lectin.

19. The method according to claim 16 wherein said capture layer assembly includes a capture agent.

20. The method according to claim 19 wherein said capture agent is a secondary capture agent.

21. The method according to claim 19 wherein said capture agent is a primary capture agent.

22. The method according to claim 19 wherein said capture agent includes a primary capture agent and a secondary capture agent.

23. The method according to claim 22 wherein said secondary capture agent binds said primary capture agent.

24. The method according to claim 23 wherein said secondary capture agent and primary capture agent is an antibody.

25. The method according to claim 22 wherein said secondary capture agent has specific affinity to said primary capture agent.

26. The method according to any one of claims 22, 23, 24, or 25 wherein said primary capture agent has specific affinity to surface markers on target cells.

27. The method according to claim 20 wherein said secondary capture agent is a secondary antibody.

28. The method according to claim 21 wherein said primary capture agent is a primary antibody.

29. The method according to claim 20 wherein said secondary capture agent is bound to said metal layer.

30. The method according to claim 21 wherein said primary capture agent is bound to said metal layer.

31. The method according to claim 15 wherein said capture layer assembly is associated with said polycarbonate substrate.

32. The method according to claim 31 wherein said capture layer assembly includes an active layer.

33. The method according to claim 32 wherein said active layer is formed from a material selected from the group comprising polystyrene, glass, nylon, streptavidin, neutravidin, avidin, biotin, and lectin.

34. The method according to claim 31 wherein said capture layer assembly includes a capture agent.

35. The method according to claim 34 wherein said capture agent is a secondary capture agent.

36. The method according to claim 34 wherein said capture agent is a primary capture agent.

37. The method according to claim 34 wherein said capture agent includes a primary capture agent and a secondary capture agent.

38. The method according to claim 37 wherein said secondary capture agent binds said primary capture agent.

39. The method according to claim 38 wherein said secondary capture agent and primary capture agent is an antibody.

40. The method according to claim 37 wherein said secondary capture agent has specific affinity to said primary capture agent.

41. The method according to any one of claims 37, 38, 39, or 40 wherein said primary capture agent has specific affinity to surface markers on target cells.

42. The method according to claim 35 wherein said secondary capture agent is a secondary antibody.

43. The method according to claim 36 wherein said primary capture agent is a primary antibody.

44. The method according to claim 35 wherein said secondary capture agent is bound to said polycarbonate substrate.

45. The method according to claim 36 wherein said primary capture agent is bound to said polycarbonate substrate.

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

providing a blood sample in a first tube, the first tube containing a separation gradient;

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

resuspending a MNC layer, the layer containing T-cells, thereby forming a MNC suspension;

providing a sample of the MNC suspension on an optical disc surface, the 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 to a 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.