Spatial positioning of spectrally labeled beads

• US 9,228,948 B2
• Filed: 04/30/2015
• Issued: 01/05/2016
• Est. Priority Date: 09/24/1998
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Empedocles, Stephen, Watson, Andrew, Jin, Jian
• Primary Examiner(s): Stock, Jr., Gordon J
• Application Number:
  US14/700,248
• Publication Number:
  US 20150247801A1
• Time in Patent Office: 250 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00574 radioactive
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00578 electrophoric
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00657 One-dimensional arrays
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2400/0454 radiation pressure, optical tweezers
• CPC Class Codes: B01L 2400/0457 : passive flow or gravitation
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: B01L 3/5085 : for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/545 : for laboratory containers
• 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 60/14 : for creating libraries
• CPC Class Codes: G01N 2015/0092 : Monitoring flocculation or agglomeration
• CPC Class Codes: G01N 2015/1472 : with colour
• CPC Class Codes: G01N 2015/1497 : Particle shape
• CPC Class Codes: G01N 2021/6441 : with two or more labels
• CPC Class Codes: G01N 2021/6482 : Sample cells, cuvettes
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/00742 : Type of codes
• CPC Class Codes: G01N 2035/00752 : bar codes
• CPC Class Codes: G01N 21/25 : Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
• CPC Class Codes: G01N 21/253 : for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/272 : for following a reaction, e.g. for determining photometrically a reaction rate (photometric cinetic analysis)
• CPC Class Codes: G01N 21/274 : Calibration, base line adjustment, drift correction
• CPC Class Codes: G01N 21/278 : Constitution of standards
• CPC Class Codes: G01N 21/6428 : Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 : Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/6456 : Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/6486 : Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 21/6489 : Photoluminescence of semiconductors
• CPC Class Codes: G01N 2201/04 : Batch operation; multisample devices
• CPC Class Codes: G01N 2201/06113 : Coherent sources; lasers
• CPC Class Codes: G01N 2201/12 : Circuits of general importance; Signal processing
• CPC Class Codes: G01N 33/54373 : involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

Devices, systems, kits, and methods for detecting and/or identifying a plurality of spectrally labeled bodies well-suited for performing multiplexed assays. By spectrally labeling the beads with materials which generate identifiable spectra, a plurality of beads may be identified within the fluid. Reading of the beads is facilitated by restraining the beads in arrays, and/or using a focused laser.

Claims

1. A method comprising:

restraining a plurality of spectrally labeled bodies with affinity binding so as to define a multiplexed array, wherein the spectrally labeled bodies are spatially resolved and comprise a plurality of fluorescent labels used to encode discrete and different emission spectra wherein each emission spectrum has a plurality of signals at differing wavelengths;

directing an image of the multiplexed array of bodies onto a sensor to sense spectra generated by the bodies; and

identifying the bodies from the spectra sensed by the sensor.

2. The method of claim 1, wherein the bodies are affixed to each other.

3. The method of claim 1, wherein the bodies are restrained within an array of openings affixed in a support structure.

4. The method of claim 1, wherein the sensor is a CCD camera.

5. A method of claim 1, wherein the spectrally labeled bodies include polymers.

6. A method of claim 1, wherein affinity binding is facilitated by a chemical moiety.

7. A method of claim 1, wherein the affinity binding is specific.

8. A method of claim 1, wherein the plurality of signals include a reference signal.

9. A method of claim 1, wherein the affinity binding is facilitated with a binding pair.

10. The method of claim 1, wherein the bodies are restrained to a support structure.

11. The method of claim 1, wherein the bodies are restrained in the multiplexed array by an array of discrete binding sites.

12. A method of claim 8, wherein the reference signal identifies a spectral code family.

13. A method of claim 9, wherein one member of the binding pair is affixed to at least one of the plurality of spectrally labeled bodies and the other member is affixed to a site of the multiplexed array.

14. The method of claim 10, wherein the support structure has a substantially planar surface.

15. The method of claim 10, wherein the support structure contains a plurality of openings sized to accommodate one of the spectrally labeled bodies.

16. The method of claim 10, wherein a plurality of unbound bodies are washed away.

17. A method of claim 11, wherein the discrete binding sites are arranged so as to inhibit a presence of more than a single body at each binding site.

18. A method of claim 11, wherein the binding sites comprise a material capable of binding to the bodies.

19. A method of claim 18, wherein the material comprises a selective affinity molecule which selectively binds to one or more particular bodies.

20. The method of claim 19, wherein the selective affinity molecule is streptavidin.

21. The method of claim 19, wherein a selective binding material is a biotinylated structure.