Two-dimensional spectral imaging system

• US 20040197816A1
• Filed: 04/26/2004
• Published: 10/07/2004
• Est. Priority Date: 04/06/2000
• Status: Abandoned Application

• Current Assignee: Invitrogen Corp. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Quantum Dot Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Empedocles, Stephen A., Watson, Andrew R.
• Application Number:
  US10/832,635
• Publication Number:
  US 20040197816A1
• Time in Patent Office: Days
• US Class Current: 435/6.13
• 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

Improved devices, systems, and methods for sensing and/or identifying signals from within a signal detection region are well-suited for identification of spectral codes. Large numbers of independently identifiable spectral codes can be generated by quite small bodies, and a plurality of such bodies or probes may be present within a detection region. Simultaneously imaging of identifiable spectra from throughout the detection region allows the probes to be identified. As the identifiable spectra can be treated as being generated from a point source within a much larger detection field, a prism, diffractive grading, holographic transmissive grading, or the like can spectrally disperse the images of the labels across a sensor surface. A CCD can identify the relative wavelengths of signals making up the spectra. Absolute signal wavelengths may be identified by determining positions of the labels, by an internal wavelength reference within the spectra, or the like.

Claims

1. A method for identifying signals of differing strengths, the method comprising:

generating a plurality of signals in response to excitation energy, the signals comprising higher intensity signals and lower intensity signals;

sensing the lower intensity signals by simultaneously imaging the signals on a sensor; and

sequentially sensing at least some of the higher intensity signals.

2. The method of claim 1, wherein at least one of the signals is generated by a semiconductor nanocrystal.

3. The method of claim 1, wherein sensing the lower intensity signals comprises imaging for a first integration time, and wherein sequentially sensing the higher intensity signals comprises sequentially imaging for a second integration time shorter than the first integration time.

4. The method of claim 1, further comprising filtering the higher intensity signals from the simultaneously imaged signals.

5. The method of claim 4, wherein the higher intensity signals have wavelengths that are different than wavelengths of the lower intensity signals, and wherein the filtering step comprises wavelength filtering the higher intensity signals.

6. The method of claim 1, wherein the higher intensity signals are sequentially sensed by scanning markers generating the signals, and wherein the markers generating the higher intensity signals are spatially intermingled with the markers generating the lower intensity signals.

7. The method of claim 6, wherein the scanning step comprises scanning an aperture relative to the markers.

8. The method of claim 7, wherein the scanning step comprises scanning a slit relative to the markers.

9. The method of claim 1, wherein the excitation energy comprises a first energy, the first energy exciting high-energy markers to generate the high energy signals, and a second energy, the second energy exciting low-energy markers to generate the lower energy signals.

10. The method of claim 9, wherein the second energy is less than the first energy, and wherein the second energy selectively excites the low energy markers.

11. The method of claim 1, wherein the high intensity signals are generated by label markers and define an identifiable spectral code, and wherein the low intensity signals are generated by assay markers and indicate results of a plurality of assays, each assay having an associated spectral code.

12. The method of claim 11, wherein the markers are supported by probe bodies to define probes, each probe comprising a label with at least one label marker to generate the spectral code, wherein at least one assay marker is associated with the probe to indicate results of an associated assay, and further comprising determining each assay result by identifying each label and correlating the label with the associated marker signal.