Methods and apparatus for detecting and quantifying lymphocytes with optical biodiscs

• US 20030104486A1
• Filed: 11/20/2001
• Published: 06/05/2003
• Est. Priority Date: 11/16/2000
• Status: Abandoned Application

• Current Assignee: Nagaoka & Co., Ltd.
• Original Assignee: Nagaoka & Co., Ltd.
• Inventors: Selvan, Gowri Pyapali
• Application Number:
  US09/988,728
• Publication Number:
  US 20030104486A1
• Time in Patent Office: Days
• US Class Current: 435/7.2
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00536 in the shape of disks
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00675 In-situ synthesis on the substrate
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00695 Synthesis control routines, e.g. using computer programs
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0806 Standardised forms, e.g. compact disc [CD] format
• CPC Class Codes: B01L 2300/168 : Specific optical properties, e.g. reflective coatings
• CPC Class Codes: B01L 2400/0409 : centrifugal forces
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• 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: B01L 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 50/14 : Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 15/1433 : using image recognition
• CPC Class Codes: G01N 2015/1486 : Counting the particles
• CPC Class Codes: G01N 21/07 : Centrifugal type cuvettes G01N21/09 takes precedence
• 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 33/56972 : White blood cells
• 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. A 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.

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. The method according to claim 1, 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.

3. The method according to claim 1, 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.

4. The method according to claim 1, 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.

5. The method according to any one of claim 1, wherein the disc surface is coated with a first group of cell capture agents.

6. The method according to claim 5, wherein the cell capture agents define a discrete capture zone.

7. The method according to claim 6, wherein a second group of cell capture agents define a second discrete capture zones in a predetermined pattern.

8. The method according to claim 7, wherein the first and second captures zones are in one chamber.

9. The method according to claim 5, wherein the cell capture agents are for binding with cell surface antigen.

10. The method according to claim 9, wherein the cell surface antigen is selected from the CD family of antigens.

11. The method according to claim 10, wherein the cell surface antigen is selected from the group consisting of CD3, CD4, CD8, and CD45.

12. The method according to claim 1, further including:

directing the sample of cells into proximity with the cell capture agents;

incubating the cells in the presence of the capture agents; and

allowing the cells to specifically bind to the capture agents.

13. The method according to claim 12, further including analyzing the number of cells captured to thereby determine a cell concentration in the sample.

14. The method of claim 13, wherein the analyzing includes detecting sufficiently large changes in the level of light reflected from or transmitted through the disc.

15. The method of claim 13, wherein the analyzing includes using image recognition to count captured cells.

16. The method of claim 15, wherein the image recognition distinguished one type of white blood cell from another.

17. The method of claim 1, wherein the chamber has a plurality of capture zones, each having a different cell capture agent.

18. The method of claim 17, wherein the rotating includes rotating for a sufficient period of time at a sufficient speed so that the cells have an opportunity to bind with the capture molecules.

19. The method of claim 18, wherein the rotating includes rotating for a sufficient period of time at a sufficient speed so that unbound cells are moved away from the capture zones.

20. The method of claim 19, wherein the rotating is done at a single speed.

21. The method of claim 17, further comprising counting the captured cells in each of the capture zones and providing an output including the counts.

22. The method of claim 21, wherein the output includes counts for CD4 cells and CD8 cells, and a ratio of CD4 to CD8 cells.

23. An optical disc comprising:

a substrate;

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

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.

24. The disc of claim 23, wherein the agents are antibodies for cell surface antigens on white blood cells.

25. The disc of claim 24, wherein the agents are antibodies for CD4 and CD8.

26. 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.

27. The disc of claim 26, wherein the detector is on the same side of the disc as the light source for detecting light reflected from the captures zones.

28. The disc of claim 26, wherein the detector is on the opposite side of the disc as the light source for detecting light transmitted through the capture zones.

29. The disc of claim 26, wherein the processor includes image recognition software for detecting imaged cells.