System for obtaining images in bright field and crossed polarization modes and chemical images in raman, luminescence and absorption modes

• US 7,564,541 B2
• Filed: 03/30/2006
• Issued: 07/21/2009
• Est. Priority Date: 06/30/2004
• Status: Active Grant

• Current Assignee: Chemimage Corporation
• Original Assignee: Chemimage Corporation
• Inventors: Tuschel, David
• Primary Examiner(s): Geisel; Kara E
• Application Number:
  US11/393,395
• Publication Number:
  US 20060170907A1
• Time in Patent Office: 1,209 Days
• Field of Search: 356 72- 73, 356/301, 356/417, 356/364, 356/445
• US Class Current: 356/73
• CPC Class Codes: G01J 3/10 Arrangements of light sources specially adapted for spectrometry or colorimetry
• CPC Class Codes: G01J 3/1256 using acousto-optic tunable filter; acousto-optic elements or systems G02F1/11, G02F1/33
• CPC Class Codes: G01J 3/26 using multiple reflection, e.g. Fabry-Perot interferometer, variable interference filters
• CPC Class Codes: G01J 3/32 Investigating bands of a spectrum in sequence by a single detector
• CPC Class Codes: G01J 3/44 Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
• CPC Class Codes: G01N 2021/6471 Special filters, filter wheel
• CPC Class Codes: G01N 2021/6493 by alternating fluorescence/transmission or fluorescence/reflection
• CPC Class Codes: G01N 21/21 Polarisation-affecting properties G01N21/19 takes precedence
• CPC Class Codes: G01N 21/23 Bi-refringence
• 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 21/35 using infrared light G01N21/39 takes precedence
• CPC Class Codes: G01N 21/3563 for analysing solids; Preparation of samples therefor
• CPC Class Codes: G01N 21/359 using near infrared light
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/65 Raman scattering

Abstract

An integrated system including one or more light sources, at least one processor, an optical lens, a two-dimensional tunable filter, one or more two-dimensional array of detection elements and instructions and a method using the integrated system. The system includes a plurality of modes: a Raman mode, an absorption mode, a luminescence mode, a crossed polarization mode, a crossed polarization absorption mode, bright field transmission or reflectance modes and a birefringence mode. The system includes instructions, executable by Sequential outputs from the two-dimensional array of detection elements is combined to generate a chemical image of the sample, wherein each of the sequential outputs from the first two-dimensional array of detection elements corresponds to spatially accurate wavelength-resolved images. The system is also used to detect dynamic changes in a sample over time by monitoring the sample using one or more of the modes.

Claims

1. A system comprising:

(a) a substantially monochromatic light source that illuminates a sample, along a first optical path, with a plurality of photons to thereby produce, along a second optical path, photons scattered or emitted from different locations on or within the sample wherein the first optical path is at an oblique angle with respect to the second optical path;

(b) a first white light source located under the sample that illuminates the sample with broadband light to thereby produce, along the second optical path, photons transmitted through the sample; and

a second white light source located above the sample that illuminates the sample with broadband light to thereby produce, along the second optical path, photons reflected from different locations on or within the sample;

(c) at least one processor that generates first control signals that control operation of the substantially monochromatic light source, the first white light source and the second white light source;

(d) an optical lens, positioned in the second optical path, that collects photons scattered, emitted, or transmitted along the second optical path;

(e) a two-dimensional tunable filter, responsive to second control signals generated by the at least one processor and positioned in the second optical path, that sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were scattered or emitted from different locations on or within the sample or transmitted through the sample;

(f) a first two-dimensional array of detection elements, responsive to third control signals generated by the at least one processor, that detects in a spatially accurate manner photons passed by the two-dimensional filter which were scattered or emitted from different locations on or within the sample or transmitted through the sample;

(g) a second two-dimensional array of detection elements, responsive to fourth control signals generated by the at least one processor, that detects in a spatially accurate manner photons reflected from different locations on or within the sample and generates an image;

(h) wherein the system includes instructions, executable by the at least one processor, for alternatively configuring the system in a Raman mode, a luminescence mode, an absorption mode and a brightfield transmission or reflection mode;

(i) wherein, in said Raman mode, the first control signals control operation of the substantially monochromatic light source in order to produce photons scattered from different locations on or within the sample, the second control signals control operation of the two-dimensional tunable filter such that the tunable filter sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were scattered from different locations on or within the sample, and the third control signals control operation of the first two-dimensional array of detection elements in order to detect in a spatially accurate manner photons passed by the two-dimensional filter which were scattered from different locations on or within the sample;

(ii) wherein, in said luminescence mode, the first control signals control operation of the substantially monochromatic light source in order to produce photons emitted from different locations on or within the sample, the second control signals control operation of the two-dimensional tunable filter such that the tunable filter sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were emitted from different locations on or within the sample, and the third control signals control operation of the first two-dimensional array of detection elements in order to detect in a spatially accurate manner photons passed by the two-dimensional filter which were emitted from different locations on or within the sample;

(iii) wherein, in said absorption mode, the first control signals control operation of the first white light source in order to produce photons transmitted through the sample, the second control signals control operation of the two-dimensional tunable filter such that the tunable filter sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were transmitted through the sample, and the third control signals control operation of the first two-dimensional array of detection elements in order to detect in a spatially accurate manner photons passed by the two-dimensional filter which were transmitted through the sample;

(iv) wherein, in said bright field mode, the first control signals control operation of one of the following;

the first white light source in order to produce photons transmitted through the sample and the second white light source in order to produce photons reflected from different locations on or within the sample, and the fourth control signals control operation of the second two-dimensional array of detection elements in order to detect in a spatially accurate manner photons which were transmitted or reflected from different locations on or within the sample and to generate a bright field image; and

(i) wherein the at least one processor combines sequential outputs from the first two-dimensional array of detection elements to generate a chemical image of the sample, wherein each of the sequential outputs from the first two-dimensional array of detection elements corresponds to spatially resolved photons in one of the plurality of predetermined wavelength bands passed by the two-dimensional tunable filter and detected by the first two-dimensional array of detection elements.

2. The system of claim 1, further comprising:

(j) a first polarizing filter located between the first white light source and the sample; and

a second polarizing filter located between the sample and the tunable filter, that receives one of transmitted or reflected photons, and is oriented 90 °

with respect to the first polarizing filter,

(k) wherein the system further includes instructions, executable by the at least one processor, for alternatively configuring the system in a crossed polarization mode;

(iv) wherein, in said crossed polarization mode, the first control signals control operation of the first white light source in order to produce photons transmitted or reflected from different locations on or within the sample, and the fourth control signals control operation of the second two-dimensional array of detection elements in order to detect in a spatially accurate manner photons which were transmitted or reflected from different locations on or within the sample and to generate a crossed polarization image.

3. The system of claim 2, wherein

(l) the system further includes instructions, executable by the at least one processor, for alternatively configuring the system in a crossed polarization absorption mode:

(i) wherein, in said crossed polarization absorption mode, the first control signals control operation of the first white light source in order to produce photons transmitted through the sample, the second control signals control operation of the two-dimensional tunable filter such that the tunable filter sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were transmitted from different locations on or within the sample, and the third control signals control operation of the first two-dimensional array of detection elements in order to detect in a spatially accurate manner photons passed by the two-dimensional filter which were transmitted through the sample.

4. The system of claim 2, further comprising a compensator located above the sample,

(l) wherein the system further includes instructions, executable by the at least one processor, for alternatively configuring the system in a birefringence mode:

(iv) wherein, in said birefringence mode, the first control signals control operation of the first white light source in order to produce photons transmitted from different locations on or within the sample, and the fourth control signals control operation of the second two-dimensional array of detection elements in order to detect in a spatially accurate manner photons which were transmitted from different locations on or within the sample and to generate a birefringence image.

5. In an integrated system that includes one or more light sources, at least one processor, an optical lens, a two-dimensional tunable filter, one or more two-dimensional array of detection elements and instructions, a method comprising,

(a) illuminating a sample, using a substantially monochromatic light source, along a first optical path, with a plurality of photons to thereby produce, along a second optical path, photons scattered or emitted from different locations on or within the sample wherein the first optical path is at an oblique angle with respect to the second optical path;

(b) illuminating the sample using at least one of the following;

a first white light source, located under the sample that illuminates the sample with broadband light to thereby produce, along the second optical path, photons transmitted through the sample or a second white light source, located above the sample that illuminates the sample with broadband light to thereby produce, along the second optical path, photons reflected from different locations on or within the sample;

(c) generating, via the at least one processor, first control signals that control operation of the substantially monochromatic light source, the first white light source and the second white light source;

(d) collecting, via the optical lens positioned in the second optical path, ones of photons scattered, emitted or transmitted along the second optical path;

(e) sequentially passing ones of photons, in each of a plurality of predetermined wavelength bands, which were scattered or emitted from different locations on or within the sample or transmitted through the sample, using the two-dimensional tunable filter, responsive to second control signals generated by the at least one processor and positioned in the second optical path;

(f) detecting, in a spatially accurate manner, ones of photons passed by the two-dimensional filter which were scattered or emitted from different locations on or within the sample or transmitted through the sample, using a first two-dimensional array of detection elements, responsive to third control signals generated by the at least one processor;

(g) optionally detecting, in a spatially accurate manner, ones of photons reflected from different locations on or within the sample, using a second two-dimensional array of detection elements, responsive to fourth control signals generated by the at least one processor and generating an image;

(h) executing instructions, by the at least one processor, for alternatively configuring the system in a Raman mode, a luminescence mode, an absorption mode and a brightfield mode;

(i) wherein, in said Raman mode, the first control signals control operation of the substantially monochromatic light source in order to produce photons scattered from different locations on or within the sample, the second control signals control operation of the two-dimensional tunable filter such that the tunable filter sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were scattered from different locations on or within the sample, and the third control signals control operation of the first two-dimensional array of detection elements in order to detect in a spatially accurate manner photons passed by the two-dimensional filter which were scattered from different locations on or within the sample;

(ii) wherein, in said luminescence mode, the first control signals control operation of the substantially monochromatic light source in order to produce photons emitted from different locations on or within the sample, the second control signals control operation of the two-dimensional tunable filter such that the tunable filter sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were emitted from different locations on or within the sample, and the third control signals control operation of the first two-dimensional array of detection elements in order to detect in a spatially accurate manner photons passed by the two-dimensional filter which were emitted from different locations on or within the sample;

(iii) wherein, in said absorption mode, the first control signals control operation of the first white light source in order to produce photons transmitted through the sample, the second control signals control operation of the two-dimensional tunable filter such that the tunable filter sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were transmitted through the sample, and the third control signals control operation of the first two-dimensional array of detection elements in order to detect in a spatially accurate manner photons passed by the two-dimensional filter which were transmitted through the sample;

(iv) wherein, in said bright field mode, the first control signals control operation of one of the following;

the first white light source in order to produce photons transmitted through the sample and the second white light source in order to produce photons reflected from different locations on or within the sample, and the fourth control signals control operation of the second two-dimensional array of detection elements in order to detect in a spatially accurate manner photons which were transmitted or reflected from different locations on or within the sample and to generate a bright field image; and

(i) combining sequential outputs, via the at least one processor, from the first two-dimensional array of detection elements to generate a chemical image of the sample, wherein each of the sequential outputs from the first two-dimensional array of detection elements corresponds to spatially resolved photons in one of the plurality of predetermined wavelength bands and detected by the first two-dimensional array of detection elements.

6. The method of claim 5, further comprising:

(j) passing the reflected or transmitted photons through a first polarizing filter located between the first white light source and the sample; and

passing the reflected or transmitted photons through a second polarizing filter located between the sample and the tunable filter and orientated 90 °

with respect to the first polarizing filter,

(k) executing instructions, by the at least one processor, for alternatively configuring the system in a crossed polarization mode;

wherein the first control signals control operation of the first white light source in order to produce photons transmitted or reflected from different locations on or within the sample, and the fourth control signals control operation of the second two-dimensional array of detection elements in order to detect in a spatially accurate manner photons which were transmitted or reflected from different locations on or within the sample and to generate a crossed polarization image.

7. The system of claim 5, further comprising:

(l) executing instructions, by the at least one processor, for alternatively configuring the system in a crossed polarization absorption mode;

wherein, in said crossed polarization absorption mode, the first control signals control operation of the first white light source in order to produce photons transmitted through the sample, the second control signals control operation of the two-dimensional tunable filter such that the tunable filter sequentially passes photons, in each of a plurality of predetermined wavelength bands, which were transmitted from different locations on or within the sample, and the third control signals control operation of the first two-dimensional array of detection elements in order to detect in a spatially accurate manner photons passed by the two-dimensional filter which were transmitted through the sample.

8. The method of claim 5, further comprising:

(l) passing transmitted photons through a compensator located above the sample;

(m) executing instructions, by the at least one processor, for alternatively configuring the system in a birefringence mode;

wherein, in said birefringence mode, the first control signals control operation of the first white light source in order to produce photons transmitted from different locations on or within the sample, and the fourth control signals control operation of the second two-dimensional array of detection elements in order to detect in a spatially accurate manner photons which were transmitted from different locations on or within the sample and to generate a birefringence image.

9. The method of claim 5 further comprising:

analyzing, by the at least one processor, the outputs from one of the following;

the first two-dimensional array of detection elements and the second two-dimensional array of detection elements, using one or more aspects of the sample that identify the sample.

10. The method of claim 9, wherein the one or more aspects of the sample comprise wavelength, frequency, phase, polarization, an intensity of wavelength, an intensity of frequency, or combinations thereof.

11. The method of claim 9, further comprising:

monitoring a change in the aspect of the sample that identifies the sample.

12. The method of claim 9, wherein the sample comprises a crystalline compound having a plurality of polymorph forms.

13. The method of claim 9, wherein the sample comprises an analyte interacting with a cell.

14. The method of claim 9, wherein the sample comprises a drug.

15. The method of claim 13, wherein the analyte comprises one or more of a drug, a metabolite of the drug, a fluorophore, or combinations thereof.