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 (d) analyzing and presenting
Type:
Grant
Filed:
July 27, 1998
Date of Patent:
March 28, 2000
Assignee:
Applied Spectral Imaging
Inventors:
Irit Bar-Am, Cathy Janish, Yuval Garini, Dario Cabib, Robert A. Buckwald
Abstract: 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 bas
Type:
Grant
Filed:
November 3, 1997
Date of Patent:
May 25, 1999
Assignee:
Applied Spectral Imaging
Inventors:
Yuval Garini, Nir Katzir, David Wine, Dario Cabib
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 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 pi
Type:
Grant
Filed:
August 25, 1997
Date of Patent:
November 10, 1998
Assignee:
Applied Spectral Imaging
Inventors:
Nir Katzir, David Wine, Yuval Garini, Dario Cabib
Abstract: 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.
Type:
Grant
Filed:
April 22, 1996
Date of Patent:
October 6, 1998
Assignee:
Applied Spectral Imaging
Inventors:
Yuval Garini, Dario Cabib, Robert A. Buckwald, Thomas Ried, Dirk G. Soenksen