Abstract:
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 so that a spectral component associated with each of the N stains is collectible. Figure (1) shows a block diagram illustrating the main components of an imaging spectrometer.
Abstract:
A method of spectral-morphometric analysis of biological samples, the biological samples including substantially constant components and suspected variable components, the method is effected by the following the steps of (a) using a spectral data collection device for collecting spectral data of picture elements of the biological samples; (b) defining a spectral vector associated with picture elements representing a constant component of at least one of the biological samples; (c) using said spectral vector for defining a correcting function being selected such that when operated on spectral vectors associated with picture elements representing other constant components, spectral vectors of said other constant components are modified to substantially resemble said spectral vector; (d) operating said correcting function on spectral associated with at least the variable components for obtaining corrected spectral vectors thereof; and (e) classifying said corrected spectral vectors into classification groups.
Abstract:
This invention is a device, and method for spectral imaging of an object. A plurality of sets of narrow-band light sources (12) such as LEDs are provided. Each set emits illumination radiation in a different narrow spectral band. Each set is activated sequentially to illuminate the object (16). Light reflected from the object or transmitted by the object is focused (26) on a detector array (20) to image the object. Narrower illumination bands are provided by dispersing the emitted light using a dispersive optical element such as a diffraction grating. Alternatively, selected sets or subsets are activated simultaneously with duty cycles that emulate a preselected spectral distribution. For imaging ocular fundus tissue, the illumination light is shaped into an annular beam by an appropriately shaped wave-guide.
Abstract:
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 (f) analyzing and presenting images of the now hybridized chromosome spread.
Abstract:
A method and an apparatus for analyzing an optical image of a scene to determine the spectral intensity of each pixel thereof, comprising: collecting and collimating incident light from the scene; passing the light through an interferometer wherein the light is first split into a finite plurality of coherent beams which travel along different optical paths inside the interferometer and then recombine to interfere with each other, the interferometer thereby outputting modulated light corresponding to a predetermined set of linear combinations of the spectral intensity of the light emitted from each pixel; focusing the light outputted from the interferometer on a two dimensional detector array; and processing the output of the detector array to determine the spectral intensity of each pixel thereof; the interferometer being of the translating type in which the optical path difference is varied to modulate the light by translating an element of the interferometer, such that at each instant each detector sees a different point of the scene and its signal is a linear combination of the spectral content of the light coming from each pixel, and that when the scanner completes one interferometer scan, the scene will have been scanned at all relevant linear combinations of the spectral content.
Abstract:
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 (114) and wide-band emission filters (116); (b) exciting fluorophores of each of the pixels with light filtered through one of the wide-band excitation filters, and recording emitted light intensity (122) 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; (d) 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.
Abstract:
A spectral imaging system comprises: a sequential optical system providing a temporal sequence of output light beams describing the scene; a color imager receiving the output light beams and responsively generating, for each output light beam, an image signal that is spatially resolved into a plurality of color channels. The system can also comprise an image processor that collectively process the image signals to construct a spectral image of the scene.
Abstract:
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 (f) analyzing and presenting images of the now hybridized chromosome spread.
Abstract:
An apparatus for use in a method of detecting and analysing fluorescent in-situ hybridisations (fig. 5) employing numerous chromosome paints (fig. 9) each labelled with a different fluorophore or combination of fluorophores, the apparatus being highly sensitive both in spatial and spectral resolutions (fig. 6) such that it is capable of simultaneous detection of dozens of fluorophores or combinations of fluorophores (fig. 7) so as to enable the detection of a complete set of fluorescently painted human chromosomes (fig. 10).