Applied Spectral Imaging Ltd.

Tel Hashomer Medical Research Infrastructure and Services Ltd.

Bar-han University

Yuval Garini

Dario Cabib

Robert A Buckwald

Nir Katzir

David Wine

Dirk G Soenksen

Moshe Lavi

Thomas Ried

George McNamara

Irit Bar-Am

Dirk G Soeknsen

Zvi Malik

Cathy Janish

Dirk C Soenksen

Irena Bronshtein-berger

Iris Barshack

A spectral imaging device includes an imager, a scanning stage to establish relative motion between the imager and a sample in a scanning direction and an optical system controlling a light characteristic of a light beam constituting an image of the sample to the imager. The optical system includes a light varying element to receive the light beam and provide an output light beam with spatially varying light characteristic over a cross-section thereof. A set of redirecting optical elements direct light rays from the sample to form the light beam, and to focus the output light beam onto the imager. A controller controls the scanning stage and the imager to capture a plurality of image frames with an overlap including a defined shift that is greater than 1 pixel along the scanning direction between consecutive image frames. A computing device consolidates image data to provide an image of the sample.

Device and method for spectral imaging

A light reflecting article is disclosed. The light reflecting article comprises a sample carrying article layered with a light reflecting layer. The light reflecting layer serves for allowing an optical collection and detection system to collect both luminescent light emitted from a sample positioned on the light reflecting article in a direction of the optical collection and detection system, as well as luminescent light emitted from the sample in a direction away from the optical collection and detection system and reflected in the direction of the optical collection and detection system via the light reflecting layer, thereby increasing a sensitivity of luminescent light detection.

Optical detection method for improved sensitivity

A method of in situ analysis of a biological sample comprising the steps of (a) staining the biological sample with N stains of which a first stain is selected from the group consisting of a first immunohistochemical stain, a first histological stain and a first DNA ploidy stain, and a second stain is selected from the group consisting of a second immunohistochemical stain, a second histological stain and a second DNA ploidy stain, with provisions that N is an integer greater than three and further that (i) if the first stain is the first immunohistochemical stain then the second stain is either the second histological stain or the second DNA ploidy stain; (ii) if the first stain is the first histological stain then the second stain is either the second immunohistochemical stain or the second DNA ploidy stain; whereas (iii) if the first stain is the first DNA ploidy stain then the second stain is either the second immunohistochemical stain or the second histological stain; and (b) using a spectral data collection device for collecting spectral data from the biological sample, the spectral data collection device and the N stains are selected such that a spectral component associated with each of the N stains is collectable.

In-situ method of analyzing cells

A spectral imaging method for simultaneous detection of multiple fluorophores aimed at detecting and analyzing fluorescent in situ hybridizations employing numerous chromosome paints and/or loci specific probes each labeled with a different fluorophore or a combination of fluorophores for color karyotyping, and at multicolor chromosome banding, wherein each chromosome acquires a specifying banding pattern, which pattern is established using groups of chromosome fragments labeled with various fluorophore or combinations of fluorophores.

Method for simultaneous detection of multiple fluorophores for in situ

A color display comprising an image of all chromosomes or portions of chromosomes of a cell, each of the chromosomes or portions of chromosomes being painted with a different fluorophore or a combination of fluorophores, the image presenting the chromosomes or portions of chromosomes in different distinctive colors, wherein each of the chromosomes or portions of chromosomes is associated with one of the different distinctive colors.

Color display of chromosomes or portions of chromosomes

A fluorescent in situ hybridization method including the steps of (a) obtaining a chromosome spread of a species; (b) preparing a hybridization composite containing a plurality of chromosomal paints each of the plurality of chromosomal paints being labeled with a different fluorophore-or-combination-of-fluorophores, such that an averaged specific activity of highly repetitive sequences in the hybridization composite substantially equals an averaged specific activity of unique sequences in the hybridization composite; (c) denaturing the hybridization composite and subjecting the hybridization composite to conditions for allowing at least a part of the highly repetitive sequences in the hybridization composite to reanneal while at least a part of the unique sequences in the hybridization composite remaining single stranded; (d) contacting under hybridization conditions the hybridization composite with the chromosome spread; (e) washing away excess of the hybridization composite; and (d) analyzing and presenting images of the now hybridized chromosome spread.

Method of and composite for in situ fluorescent hybridization

A method for cancer cell detection including the steps of (a) staining an analyzed sample with at least first and second dyes, the dyes being selected such that the first dye better adheres to normal cells whereas the second dye better adheres to cancer cells; (b) spectrally imaging the sample through an optical device being optically connected to an imaging spectrometer thereby obtaining a spectrum of each pixel of the sample; (c) based on the spectra, evaluating concentrations of the first and second dyes for each of the pixels; and (d) based on the concentrations detecting the presence of cancer cells in the sample.

Method of cancer cell detection

A fluorescent in situ hybridization method comprising the steps of (a) providing a cell nuclei having chromosomes hybridized with at least one nucleic acid probe including at least one nucleic acid molecule labeled with at least one fluorophore; (b) viewing the cell nuclei through a fluorescence microscope optically connected to an imaging spectrometer for obtaining a spectrum of each pixel of the cell nuclei by (i) collecting incident collimated light simultaneously from all pixels of the cell nuclei; (ii) passing the incident collimated light through an interferometer system so that the light is first split into two coherent beams and then recombine to interfere and form an exiting light beam; (iii) focusing the exiting light beam on a detector having an array of detector elements, so that at each instant each of the elements is the image of one and always the same pixel for the entire duration of the measurement and so that each of the elements produces a signal which is a particular linear combination of light intensity emitted by the pixel at different wavelengths; (iv) rotating or translating one or more of the elements of the interferometer, so that the optical path difference is scanned simultaneously for all the pixels of the cell nuclei; 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.

A method for finding L internal reference vectors for classification of L chromosomes or portions of chromosomes of a cell, the L chromosomes or portions of chromosomes being painted with K different fluorophores or combinations thereof, wherein K basic chromosomes or portions of chromosomes of the L chromosomes or portions of chromosomes are each painted with only one of the K different fluorophores, whereas the other L-K of the L chromosomes or portions of chromosomes are each painted with a different combination of the K different fluorophores, the method comprising the steps of (a) using a multi-band collection device for measuring a first vector for each pixel of each of the L chromosomes or portions of chromosomes; (b) identifying pixels belonging to each of the K basic chromosomes or portions of chromosomes and defining the pixels as basic pixels, so as to obtain K basic classes of basic pixels; (c) using at least one basic pixel from each of the K basic classes for obtaining K basic vectors, the K basic vectors being K internal reference vectors; (d) using the K basic vectors for identifying pixels belonging to the other L-K chromosomes or portions of chromosomes; and (e) using the pixels belonging to the other L-K chromosomes or portions of chromosomes for calculating the other L-K internal reference vectors, thereby finding all of the L internal reference vectors. A method for classification of L chromosomes or portions of chromosomes of a cell similarly painted using the above method for finding L internal reference vectors, and using the L reference vectors for classification of each of the pixels into one of L classification classes. And, images presenting color chromosomes.

Method for chromosomes classification

A fluorescent in situ hybridization method including the steps of (a) obtaining a chromosome spread of a species; (b) preparing a hybridization composite containing a plurality of chromosomal paints each of the plurality of chromosomal paints being labeled with a different fluorophore-or-combination-of-fluorohores, such that an averaged specific activity of highly repetitive sequences in the hybridization composite substantially equals an averaged specific activity of unique sequences in the hybridization composite; (c) denaturing the hybridization composite and subjecting the hybridization composite to conditions for allowing at least a part of the highly repetitive sequences in the hybridization composite to reanneal while at least a part of the unique sequences in the hybridization composite remaining single stranded; (d) contacting under hybridization conditions the hybridization composite with the chromosome spread; (e) washing away excess of the hybridization composite; and (d) analyzing and presenting images of the now hybridized chromosome spread.

Method of and composite for fluorescent in situ hybridization

A method of classification of pixels into groups of pixels according to their association with a single fluorophore or a combination of fluorophores selected from a plurality of fluorophores, each of the fluorophores having characterizing excitation and emission spectra and specifying excitation and emission peaks, the method comprising the steps of (a) providing a plurality of pairs of wide-band excitation filters and wide-band emission filters; (b) exciting fluorophores of each of the pixels with light filtered through one of the wide-band excitation filters, and recording emitted light intensity as retrieved after passing through its paired emission filter; (c) repeating step (b) for all of the plurality of pairs of filters, such that each of the pixels is representable by a vector of a plurality of dimensions, the number of dimensions being equal to the number of the plurality of pairs of filters; (c) using an algorithm for evaluating the presence of each of the plurality of fluorophores in each of the pixels, thereby classifying each of the pixels into a group of pixels according to its association with a single fluorophore or combination of fluorophores.

Method for classification of pixels into groups according to their

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.

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Koranit, Israel

Yuval Garini