Two-dimensional spectral imaging system

• US 20070035819A1
• Filed: 06/30/2006
• Published: 02/15/2007
• Est. Priority Date: 06/30/2005
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Prothro, Derek, Patel, Chirag, Bahatt, Dar, Tan, Roy
• Granted Patent: US 7,663,750 B2
• Time in Patent Office: Days
• US Class Current: 359/366
• CPC Class Codes: G01J 2003/1204 Grating and filter
• CPC Class Codes: G01J 2003/1208 Prism and grating
• CPC Class Codes: G01J 2003/1213 Filters in general, e.g. dichroic, band
• CPC Class Codes: G01J 2003/1226 Interference filters
• CPC Class Codes: G01J 3/10 Arrangements of light sources specially adapted for spectrometry or colorimetry
• CPC Class Codes: G01J 3/12 Generating the spectrum; Monochromators
• CPC Class Codes: G01J 3/18 using diffraction elements, e.g. grating gratings per se G02B
• CPC Class Codes: G01J 3/189 using at least one grating in an off-plane configuration
• CPC Class Codes: G01J 3/2803 using photoelectric array detector
• CPC Class Codes: G01J 3/2823 Imaging spectrometer
• CPC Class Codes: G01J 3/4406 Fluorescence spectrometry
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6432 Quenching
• CPC Class Codes: G01N 21/0332 with temperature control control of temperature G05D23/00; cryostats F17C3/08
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/31 Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry G01N21/72 takes precedence
• 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 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 2201/06113 : Coherent sources; lasers
• CPC Class Codes: G01N 2201/0621 : Supply
• CPC Class Codes: G01N 2201/0697 : Pulsed lasers

Abstract

Systems, including methods, apparatus, and algorithms, for spectrally imaging a two-dimensional array of samples.

Claims

1. A system for sample analysis, comprising:

an examination area including a two-dimensional array of examination sites;

a light detector for imaging;

an optical relay structure that spectrally disperses light received from the examination sites into a corresponding two-dimensional array of spectra directed concurrently onto the light detector in a collectively nonoverlapping relation with nondispersed light from the two-dimensional array of examination sites; and

a thermal control system operatively coupled to the examination area for regulating a temperature thereof.

2. The system of claim 1, further comprising a controller, wherein the thermal control system is operatively coupled to the controller for automated thermal control of the examination area.

3. The system of claim 1, wherein the thermal control system is configured to repetitively follow a thermal profile in an automated manner.

4. The system of claim 1, wherein the thermal control system includes a heater and a thermally conductive member operatively coupled to the heater, further comprising a sample holder having wells and configured to be disposed in the examination area such that the wells are disposed in the examination sites and the sample holder is in contact with the conductive member.

5. The system of claim 4, wherein the conductive member and the sample holder have complementary structure that restricts lateral movement of the sample holder when the sample holder is disposed in the examination area.

6. The system of claim 5, wherein the complementary structure includes one or more projections of the conductive member that are configured to be received in one or more openings of the sample holder.

7. The system of claim 1, further comprising a sample holder having wells and configured to be disposed in the examination area such that the wells are disposed in the examination sites, wherein the wells have a staggered arrangement.

8. The system of claim 7, wherein the wells are disposed along a set of spaced parallel lines with a well-to-well spacing within each line, and wherein every other line of wells is offset along its line by about one-half the well-to-well spacing.

9. The system of claim 1, further comprising a sample holder having wells and configured to be disposed in the examination area such that the wells are disposed in the examination sites, wherein each well is at least about one millimeter in diameter.

10. The system of claim 1, further comprising a sample holder having wells and configured to be disposed in the examination area such that the wells are disposed in the examination sites, wherein each well has a capacity of at least about 100 nanoliters.

11. The system of claim 1, wherein the two-dimensional array of spectra defines orthogonal axes, and wherein each spectrum extends along a spectral axis that is oblique to each of the orthogonal axes.

12. The system of claim 1, further comprising a controller operatively coupled to the light detector and configured to process image data collected by the light detector to correct for optical distortion in the image data.

13. A method of sample analysis, comprising:

obtaining a two-dimensional array of samples;

illuminating each sample of the array such that the sample emits light;

directing light from each sample to a detection area such that a corresponding two-dimensional array of spectra falls concurrently onto the detection area;

detecting the two-dimensional array of spectra repeatedly over time to measure a change, if any, in each spectrum; and

determining an aspect of one or more of the samples based on a measured change in one or more corresponding spectra.

14. The method of claim 13, wherein the step of directing includes a step of directing light such that the two-dimensional array of spectra collectively is resolved from nondispersed light from the samples.

15. The method of claim 14, wherein the step of directing includes a step of directing nondispersed light from one or more of the samples onto the detection area.

16. The method of claim 13, wherein the step of obtaining a two-dimensional array of samples includes a step of obtaining a two-dimensional array of reaction mixtures for performing amplification of nucleic acids.

17. A method of sample analysis, comprising:

obtaining a two-dimensional array of samples for performing amplification of nucleic acids;

illuminating each sample of the array concurrently such that the samples emit light;

directing dispersed light from each sample to a detection area such that a corresponding two-dimensional array of spectra falls concurrently onto the detection area;

monitoring the two-dimensional array of spectra over time to measure a change, if any, in each spectrum; and

determining an aspect of one or more of the samples based on a measured change in one or more corresponding spectra.

18. The method of claim 17, further comprising a step of cycling the samples thermally through a plurality of cycles.

19. A system for sample analysis, comprising:

a sample holder including a two-dimensional array of wells for receiving samples;

an examination area in which the sample holder is disposed;

a light detector for imaging; and

an optical relay structure operatively coupled to the examination area and configured to spectrally disperse light received from the wells parallel to a dispersion axis into a corresponding two-dimensional array of spectra directed concurrently onto the light detector in a collectively nonoverlapping relation with nondispersed light from the wells,

wherein each well has a size sufficient to substantially broaden its corresponding spectrum parallel to the dispersion axis relative to a spectrum of the same spectral composition produced from a point source in place of the well.

20. The system of claim 19, wherein the spectrum from each well is broadened by at least about ten nanometers relative to the spectrum produced from a point source in place of the well.

21. The system of claim 19, wherein each well has a diameter of at least about one millimeter.

22. The system of claim 19, wherein the optical relay structure includes one or more filters that select a range of wavelengths of the dispersed light to direct onto the light detector.

23. The system of claim 22, wherein the range of wavelengths at least substantially excludes visible light at opposing ends of a full-length visible spectrum.

24. A system for sample analysis, comprising:

a sample holder including a two-dimensional array of wells and one or more sealing elements configured to seal each of the wells after the wells have received samples;

an examination area in which the sample holder is positioned;

a light detector for imaging; and

an optical relay structure that, when the sample holder is positioned in the examination area, spectrally disperses light received from the two-dimensional array of wells into a corresponding two-dimensional array of spectra directed concurrently onto the light detector in a collectively nonoverlapping relation with nondispersed light from the two-dimensional array of wells.