US6066459A
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Method for simultaneous detection of multiple fluorophores for in situ hybridization and multicolor chromosome painting and banding
Assignee: Applied Spectral Imaging Ltd
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Filing: 1998-06-19
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Granted: 2000-05-23
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Expires: 2018-06-19
Abstract
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.
Inventors
- 👤 Yuval Garini
- 👤 Dario Cabib
- 👤 Robert A. Buckwald
- 👤 Thomas Ried
- 👤 Dirk G. Soenksen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 10:19
CPC Classifications
G; G01; G01N; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/645; G01N21/6452; C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6841; C12Q1/6876; C12Q1/6883; G01J; G01J3/00; G01J3/12; G01J3/1256; G01J3/26; G01J3/28; G01J3/2823; G01J3/44; G01J3/4406; G01J3/45; G01J3/453; G01N21/6428; G01N21/6456; G01N21/6458; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/53; G01N33/569; G01N33/56966; G01N33/58; G01N33/582; G06; G06V; G06V10/00; G06V10/88; G06V20/00; G06V20/60; G06V20/69; C12Q2600/00; C12Q2600/156; G01J2003/2866; G01J3/02; G01N2021/6417; G01N2021/6439; G01N2021/6441
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
188 words
A method of hybridization based multicolor chromosome banding comprising the steps of: (a) providing a metaphase chromosome spread of a first species and chromosome paints of a second species, at least some of said chromosome paints being labeled with a different fluorophore or combination of fluorophores; (b) hybridizing said chromosome paints with said metaphase chromosome spread; (c) using a fluorescence microscope being optically coupled to a multiband collection device for collecting spectral data from said chromosome spread; and (d) based on said collected data calculating a color image in which pixels associated with each of said different fluorophores or combination of fluorophores are attributed distinctive colors, thereby a banded color image of said metaphase chromosome spread is obtainable; wherein said method is for an application selected from the group consisting of providing a color banding karyotype of embryonic cells, providing a color banding karotype of white blood cells, providing a color banding karyotype of malignant cells, providing a color banding karyotype of cells examined for malignancy, detecting the stage of a malignancy, following the efficiency of an anti-cancer treatment and detecting chromosomal aberrations following exposure to mitogenic agents.
Claim 2
Independent Claim
188 words
A method of hybridization based multicolor chromosome banding comprising the steps of: (a) providing a metaphase chromosome spread of a first species and chromosome paints of a second species, at least some of said chromosome paints being labeled with a different fluorophore or combination of fluorophores; (b) hybridizing said chromosome paints with said metaphase chromosome spread; (c) using a fluorescence microscope being optically coupled to a multiband collection device for collecting spectral data from said chromosome spread; and (d) based on said collected data calculating a color image in which pixels associated with each of said different fluorophores or combination of fluorophores are attributed distinctive colors, thereby a banded color image of said metaphase chromosome spread is obtainable; wherein said method is for an application selected from the group consisting of providing a color banding karyotype of embryonic cells, providing a color banding karotype of white blood cells, providing a color banding karyotype of malignant cells, providing a color banding karyotype of cells examined for malignancy, detecting the stage of a malignancy, following the efficiency of an anti-cancer treatment and detecting chromosomal aberrations following exposure to mitogenic agents.
Claim 3
Dependent Claim
33 words
A method as in claim 1, where in said fluorescence microscope is supplemented with a filter selected from the group consisting of a double band dichroic filter and a triple band dichroic filter.
Claim 4
Dependent Claim
26 words
A method as in claim 1, wherein said chromosome paints are labeled using a strategy selected from the group consisting of combinatorial labeling and combinatorial hybridization.
Claim 5
Dependent Claim
16 words
A method as in claim 1, wherein said chromosome paints are labeled by interspersed repetitive sequence--PCR.
Claim 6
Dependent Claim
18 words
A method as in claim 1, wherein said fluorophores are conjugated to a nucleotide or a nucleotide analog.
Claim 7
Dependent Claim
25 words
A method as in claim 1, wherein said first and second species are each independently selected from the group consisting of, human, mouse and monkey.
Claim 8
Dependent Claim
32 words
A method as in claim 1, wherein said embryonic cells are selected from the group consisting of chorionic villi cells and embryonic cells isolated from the peripheral blood of a pregnant woman.
Claim 9
Dependent Claim
57 words
A method as in claim 7, wherein said method is for detecting a trisomy of a genetic material selected from the group consisting of human chromosome 21, human chromosomal band 21q22, a fragment of human chromosomal band 21q22, human chromosome 18, a fragment of human chromosome 18, human chromosome 13 and a fragment of human chromosome 13.
Claim 10
Dependent Claim
26 words
A method as in claim 7, wherein said providing said color banding karyotype of said cells examined for malignancy, is for obtaining a color translocation map.
Claim 11
Dependent Claim
23 words
A method as in claim 1, wherein said detection of the stage of said malignancy is for selecting a treatment for said malignancy.
Claim 12
Dependent Claim
35 words
A method as in claim 1, wherein said metaphase chromosome spread is stained with a conventional chromosome banding dye, the method further comprising the step of obtaining a gray level banding image of said chromosomes.
Claim 13
Dependent Claim
14 words
A method as in claim 11, wherein said conventional chromosome banding dye is DAPI.
Claim 14
Dependent Claim
15 words
A method as in claim 1, wherein said multiband collection device is an imaging spectrometer.
References Cited (Backward Citations)
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