US 5,784,162 A · Spectral bio-imaging methods for biological research, medical diagnostics and therapy

Abstract

According to the present invention there are provided spectral imaging methods for biological research, medical diagnostics and therapy comprising the steps of (a) preparing a sample to be spectrally imaged; (b) viewing the sample through an optical device, the optical device being optically connected to an imaging spectrometer, the optical device and the imaging spectrometer obtaining a spectrum of each pixel of the sample by: (i) collecting incident light simultaneously from all pixels of the sample using collimating optics; (ii) passing the incident collimated light through an interferometer system having a number of elements, to form an exiting light beam; (iii) passing the exiting light beam through a focusing optical system which focuses the exiting light beam on a detector having a two-dimensional array of detector elements, so that at each instant each of the detector elements is the image of one pixel of the sample, so that the real image of the sample is stationary on the plane of the detector array, and so that each of the detector elements produces a signal which is a particular linear combination of light intensity emitted by the pixel at different wavelengths, wherein the linear combination is a function of the instantaneous optical path difference; (iv) rotating one or more of the elements of the interferometer system, so that the optical path difference between the two coherent beams generated by the interferometer system is scanned simultaneously for all the pixels of the sample; and (v) recording signals of each of the detector elements as function of time using a recording device to form a first spectral cube of data; and (c) interpreting the first spectral cube of data using a mathematical algorithm.

Claims69

Dependent claims

A spectral bio-imaging method characterized by high spatial and high spectral resolutions, the method comprising the steps of:

(a) preparing a sample to be spectrally imaged;
(b) viewing said sample through an optical device, said optical device being optically connected to an imaging spectrometer, said optical device and said imaging spectrometer being for obtaining a spectrum of each pixel of said sample by:
(i) collecting incident light simultaneously from all pixels of said sample using collimating optics;
(ii) passing said incident collimated light through an interferometer system having a number of elements, so that said light is first split into two coherent beams which travel in different directions inside said interferometer and then said two coherent beams recombine to interfere with each other to form an exiting light beam;
(iii) passing said exiting light beam through a focusing optical system which focuses said exiting light beam on a detector having a two-dimensional an-ay of detector elements, so that at each instant each of said detector elements is the image of one and always the same pixel of said sample for the entire duration of the measurement, so that the real image of the sample is stationary on the plane of the detector array and at any time during the measurement the image is still visible and recognizable, and so that each of said detector elements produces a signal which is a particular linear combination of light intensity emitted by said pixel at different wavelengths, wherein said linear combination is a function of the instantaneous optical path difference;
(iv) rotating one or more of said elements of said interferometer system, so that said optical path difference between said two coherent beams generated by said interferometer system is scanned simultaneously for all said pixels of said sample;
and
(v) recording signals of each of said detector elements as function of time using a recording device to form a first spectral cube of data;
and
(c) interpreting said first spectral cube of data using a mathematical algorithm.

7 more elements

A fluorescent in situ hybridization method comprising the steps of:

(a) labeling with at least one fluorescent dye at least one nucleic acid molecule to obtain at least one fluorescently tagged nucleic acid probe;
(b) hybridizing said probe in situ with cellular nucleic acids of a biological sample;
(c) viewing said biological sample through a fluorescence microscope, said fluorescence microscope being optically connected to an imaging spectrometer, said fluorescence microscope and said imaging spectrometer being for obtaining a spectrum of each pixel of said biological sample by:
(i) collecting incident light simultaneously from all pixels of said biological sample using collimating optics;
(ii) passing said incident collimated light through an interferometer system having a number of elements, so that said light is first split into two coherent beams which travel in different directions inside said interferometer and then said two coherent beams recombine to interfere with each other to form an exiting light beam;
(iii) passing said exiting light beam through a focusing optical system which focuses said exiting light beam on a detector having a two-dimensional allay of detector elements, so that at each instant each of said detector elements is the image of one and always the same pixel of said biological sample for the entire duration of the measurement, so that the real image of the biological sample is stationary on the plane of the detector array and at any time during the measurement the image is still visible and recognizable, and so that each of said detector elements produces a signal which is a particular linear combination of light intensity emitted by said pixel at different wavelengths, wherein said linear combination is a function of the instantaneous optical path difference;
(iv) rotating one or more of said elements of said interferometer system, so that said optical path difference between said two coherent beams generated by said interferometer system is scanned simultaneously for all said pixels of said biological sample;
and
(v) recording signals of each of said detector elements as function of time using a recording device to form a first spectral cube of data;
and
(d) interpreting said first spectral cube of data using a mathematical algorithm.

8 more elements

A fluorescent in situ hybridization method comprising the steps of:

(a) hybridizing at least one nucleic acid probe in situ with cellular nucleic acids of a biological sample;
(b) labeling each of said at least one probe with at least one fluorescent dye;
(c) viewing said biological sample tough a fluorescence microscope, said fluorescence microscope being optically connected to an imaging spectrometer, said fluorescence microscope and said imaging spectrometer being for obtaining a spectrum of each pixel of said biological sample by:
(i) collecting incident light simultaneously from all pixels of said biological sample using collimating optics;
(ii) passing said incident collimated light through an interferometer system having a number of elements, so that said light is first split into two coherent beams which travel in different directions inside said interferometer and then said two coherent beams recombine to interfere with each other to form an exiting light beam;
(iii) passing said exiting light beam through a focusing optical system which focuses said exiting light beam on a detector having a two-dimensional array of detector elements, so that at each instant each of said detector elements is the image of one and always the same pixel of said biological sample for the entire duration of the measurement, so that the real image of the biological sample is stationary on the plane of the detector array and at any time during the measurement the image is still visible and recognizable, and so that each of said detector elements produces a signal which is a particular linear combination of light intensity emitted by said pixel at different wavelengths, wherein said linear combination is a function of the instantaneous optical path difference;
(iv) rotating one or more of said elements of said interferometer system, so that said optical path difference between said two coherent beams generated by said interferometer system is scanned simultaneously for all said pixels of said biological sample;
and
(v) recording signals of each of said detector elements as function of time using a recording device to form a first spectral cube of data;
and
(d) interpreting said first spectral cube of data using a mathematical algorithm.

8 more elements

A cell classification method comprising the steps of:

(a) preparing a smear of cells for analysis;
(b) viewing said smear of cells through a transmission microscope, said transmission microscope being optically connected to an imaging spectrometer, transmission microscope and said imaging spectrometer being for obtaining a spectrum of each pixel of said smear of cells by:
(i) collecting incident light simultaneously from all pixels of said smear of cells using collimating optics;
(ii) passing said incident collimated light through an interferometer system having a number of elements, so that said light is first split into two coherent beams which travel in different directions inside said interferometer and then said two coherent beams recombine to interfere with each other to form an exiting light beam;
(iii) passing said exiting light beam through a focusing optical system which focuses said exiting light beam on a detector having a two-dimensional array of detector elements, so that at each instant each of said detector elements is the image of one and always the same pixel of said smear of cells for the entire duration of the measurement, so that the real image of the smear of cells is stationary on the plane of the detector array and at any time during the measurement the image is still visible and recognizable, and so that each of said detector elements produces a signal which is a particular linear combination of light intensity emitted by said pixel at different wavelengths, wherein said linear combination is a function of the instantaneous optical path difference;
(iv) rotating one or more of said elements of said interferometer system, so that said optical path difference between said two coherent beams generated by said interferometer system is scanned simultaneously for all said pixels of said smear of cells;
and
(v) recording signals of each of said detector elements as function of time using a recording device to form a first spectral cube of data;
and
(c) interpreting said first spectral cube of data using a principal component algorithm.

7 more elements

Dependencies are inferred from the claim text.

Spectral bio-imaging methods for biological research, medical diagnostics and therapy

Life of this patent

Spectral bio-imaging methods for biological research, medical diagnostics and therapy

Life of this patent

Abstract

Claims69