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US6007996A
Expired / Lapsed
US
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In situ method of analyzing cells
Assignee: Applied Spectral Imaging Ltd
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Filing: 1998-07-27
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Granted: 1999-12-28
•
Expires: 2018-07-27
Abstract
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 such that a spectral component associated with each of the N stains is collectable.
Inventors
- 👤 George McNamara
- 👤 Dirk Soenksen
- 👤 Dario Cabib
- 👤 Robert A. Buckwald
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 23:07
CPC Classifications
G; G01; G01N; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/645; G01N21/6456; G01N21/6458; 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; G01N21/6486; 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; G01J2003/2866; G01J3/02; G01N2021/6417; G01N2021/6423
Claims Analysis
Total Claims: 69
Claim 1
Independent Claim
206 words
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 said first stain is said first immunohistochemical stain then said second stain is either said second histological stain or said second DNA ploidy stain; (ii) if said first stain is said first histological stain then said second stain is either said second immunohistochemical stain or said second DNA ploidy stain; whereas (iii) if said first stain is said first DNA ploidy stain then said second stain is either said second immunohistochemical stain or said second histological stain; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said N stains are selected such that a spectral component associated with each of said N stains is collectable.
Claim 2
Independent Claim
206 words
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 said first stain is said first immunohistochemical stain then said second stain is either said second histological stain or said second DNA ploidy stain; (ii) if said first stain is said first histological stain then said second stain is either said second immunohistochemical stain or said second DNA ploidy stain; whereas (iii) if said first stain is said first DNA ploidy stain then said second stain is either said second immunohistochemical stain or said second histological stain; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said N stains are selected such that a spectral component associated with each of said N stains is collectable.
Claim 3
Dependent Claim
37 words
The method of claim 1, wherein said spectral data collection device is selected from the group consisting of an interferometer-based spectral data collection device, filters-based spectral data collection device and a dispersion element-based spectral data collection device.
Claim 4
Dependent Claim
18 words
The method of claim 1, wherein said first and second immunohistochemical stains each independently includes a primary antibody.
Claim 5
Dependent Claim
159 words
The method of claim 3, wherein said primary antibody is selected from the group consisting of anti-estrogen receptor antibody, anti-progesterone receptor antibody, anti-p53 antibody, anti-Her-2/neu antibody, anti-EGFR antibody, anti-cathepsin D antibody, anti-Bcl-2 antibody, anti-E-cadherin antibody, anti-CA125 antibody, anti-CA15-3 antibody, anti-CA19-9 antibody, anti-c-erbB-2 antibody, anti-P-glycoprotein antibody, anti-CEA antibody, anti-retinoblastoma protein antibody, anti-ras oncoprotein antibody, anti-Lewis X antibody, anti-Ki-67 antibody, anti-PCNA antibody, anti-CD3 antibody, anti-CD4 antibody, anti-CD5 antibody, anti-CD7 antibody, anti-CD8 antibody, anti-CD9/p24 antibody, anti-CD10 antibody, anti-CD11c antibody, anti-CD13 antibody, anti-CD14 antibody, anti-CD15 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD23 antibody, anti-CD30 antibody, anti-CD31 antibody, anti-CD33 antibody, anti-CD34 antibody, anti-CD35 antibody, anti-CD38 antibody, anti-CD41 antibody, anti-LCA/CD45 antibody, anti-CD45RO antibody, anti-CD45RA antibody, anti-CD39 antibody, anti-CD100 antibody, anti-CD95/Fas antibody, anti-CD99 antibody, anti-CD106 antibody, anti-ubiquitin antibody, anti-CD71 antibody, anti-c-myc antibody, anti-cytokeratins antibody, anti-vimentins antibody, anti-HPV proteins antibody, anti-kappa light chains antibody, anti-lambda light chains antibody, anti-melanosomes antibody, anti-prostate specific antigen antibody, anti-S-100 antibody, anti-tau antigen antibody, anti-fibrin antibody, anti-keratins antibody and anti-Tn-antigen antibody.
Claim 6
Dependent Claim
15 words
The method of claim 3, wherein said primary antibody is labeled with a detectable moiety.
Claim 7
Dependent Claim
13 words
The method of claim 5, wherein said detectable moiety is a fluorescent dye.
Claim 8
Dependent Claim
120 words
The method of claim 6, wherein said fluorescent dye is selected from the group consisting of Fluorescein, Rhodamine, Texas Red, Cy2, Cy3, Cy5, VECTOR Red, ELF™ (Enzyme-Labeled Fluorescence), Cy0, Cy0.5, Cy1, Cy1.5, Cy3, Cy3.5, Cy5, Cy7, FluorX, Calcein, Calcein-AM, CRYPTOFLUOR™'S, Orange (42 kDa), Tangerine (35 kDa), Gold (31 kDa), Red (42 kDa), Crimson (40 kDa), BHMP, BHDMAP, Br-Oregon, Lucifer Yellow, Alexa dye family, N-[6-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino]caproyl] (NBD), BODIPY™, boron dipyrromethene difluoride, Oregon Green, MITOTRACKER™ Red, DiOC 7 (3), DiIC 18 , Phycoerythrin, Phycobiliproteins BPE (240 kDa) RPE (240 kDa) CPC (264 kDa) APC (104 kDa), Spectrum Blue, Spectrum Aqua, Spectrum Green, Spectrum Gold, Spectrum Orange, Spectrum Red, NADH, NADPH, FAD, Infra-Red (IR) Dyes, Cyclic GDP-Ribose (cGDPR), Calcofluor White, Tyrosine and Tryptophan.
Claim 9
Dependent Claim
13 words
The method of claim 5, wherein said detectable moiety is a non-fluorescent dye.
Claim 10
Dependent Claim
13 words
The method of claim 8, wherein said non-fluorescent dye includes a heavy metal.
Claim 11
Dependent Claim
26 words
The method of claim 5, wherein said detectable moiety is an enzyme catalyzing a colorimetric reaction of a substrate having a substantially non-soluble color reaction product.
Claim 12
Dependent Claim
23 words
The method of claim 10, wherein said enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase, and glucose oxidase.
Claim 13
Dependent Claim
27 words
The method of claim 10, wherein said substrate is selected from the group consisting of alkaline phosphatase substrates, horseradish peroxidase substrates, glucose oxidase substrates and beta-galactosidase substrates.
Claim 14
Dependent Claim
49 words
The method of claim 5, wherein said detectable moiety is an enzyme catalyzing a luminescence reaction of a substrate having a substantially non-soluble reaction product capable of luminescencing or of chemical involvement in a second reaction with at least one additional substrate, said second reaction having a luminescencing product.
Claim 15
Dependent Claim
23 words
The method of claim 13, wherein said enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase and glucose oxidase.
Claim 16
Dependent Claim
32 words
The method of claim 13, wherein said first and at least one additional substrates are each independently selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase and glucose oxidase substrates.
Claim 17
Dependent Claim
23 words
The method of claim 1, wherein said first and second immunohistochemical stains each independently includes a primary antibody and a signal amplification mechanism.
Claim 18
Dependent Claim
55 words
The method of claim 16, wherein said signal amplification mechanism is selected from the group consisting of a secondary antibody capable of binding a constant region of said primary antibody, avidin or strepavidin capable of binding biotin conjugated to said primary antibody and biotin capable of binding avidin or strepavidin conjugated to said primary antibody.
Claim 19
Dependent Claim
21 words
The method of claim 14, wherein said secondary antibody, avidin, strepavidin and biotin are each independently labeled with a detectable moiety.
Claim 20
Dependent Claim
13 words
The method of claim 18, wherein said detectable moiety is a fluorescent dye.
Claim 21
Dependent Claim
120 words
The method of claim 19, wherein said fluorescent dye is selected from the group consisting of Fluorescein, Rhodamine, Texas Red, Cy2, Cy3, Cy5, VECTOR Red, ELF™ (Enzyme-Labeled Fluorescence), Cy0, Cy0.5, Cy1, Cy1.5, Cy3, Cy3.5, Cy5, Cy7, FluorX, Calcein, Calcein-AM, CRYPTOFLUOR™'S, Orange (42 kDa), Tangerine (35 kDa), Gold (31 kDa), Red (42 kDa), Crimson (40 kDa), BHMP, BHDMAP, Br-Oregon, Lucifer Yellow, Alexa dye family, N-[6-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino]caproyl] (NBD), BODIPY™, boron dipyrromethene difluoride, Oregon Green, MITOTRACKER™ Red, DiOC 7 (3), DiIC 18 , Phycoerythrin, Phycobiliproteins BPE (240 kDa) RPE (240 kDa) CPC (264 kDa) APC (104 kDa), Spectrum Blue, Spectrum Aqua, Spectrum Green, Spectrum Gold, Spectrum Orange, Spectrum Red, NADH, NADPH, FAD, Infra-Red (IR) Dyes, Cyclic GDP-Ribose (cGDPR), Calcofluor White, Tyrosine and Tryptophan.
Claim 22
Dependent Claim
13 words
The method of claim 18, wherein said detectable moiety is a non-fluorescent dye.
Claim 23
Dependent Claim
25 words
The method of claim 21, wherein said non-fluorescent dye is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, glucose oxidase and beta-galactosidase substrates.
Claim 24
Dependent Claim
26 words
The method of claim 18, wherein said detectable moiety is an enzyme catalyzing a colorimetric reaction of a substrate having a substantially non-soluble color reaction product.
Claim 25
Dependent Claim
23 words
The method of claim 23, wherein said enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase, and glucose oxidase.
Claim 26
Dependent Claim
24 words
The method of claim 23, wherein said substrate is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase, and glucose oxidase substrates.
Claim 27
Dependent Claim
42 words
The method of claim 18, wherein said detectable moiety is an enzyme catalyzing a luminescence reaction of a substrate having a substantially non-soluble reaction product capable of luminescencing or of directing a second reaction of a second substrate having a luminescencing product.
Claim 28
Dependent Claim
18 words
The method of claim 26, wherein said enzyme is selected from the group consisting of luciferase and aequorin.
Claim 29
Dependent Claim
27 words
The method of claim 26, wherein said first and second substrates are each independently selected from the group consisting of luciferine, ATP, coelenterazine and Ca ++ .
Claim 30
Dependent Claim
66 words
The method of claim 1, wherein said first and second histological stains are each independently selected from the group consisting of 4',6-diamidino-2-phenylindole, Eosin, Fluorescein isothiocyanate, Hoechst 33258, Hoechst 33342, Propidium Iodide, Quinacrine, Fluorescein-phalloidin, Resorufin, hematoxylin, Orange G, Light Green SF, Romanowsky-Giemsa, May-Grunwald, Blue counterstain, ethyl green, Feulgen-naphthol yellow S, Giemsa, Methylene Blue, Methyl Green, pyronin, Naphthol-yellow, Neutral Red, Papanicolaou stain, Red Counterstain C and Sirius Red.
Claim 31
Dependent Claim
68 words
The method of claim 1, wherein said first and second DNA ploidy stains are each independently selected from the group consisting of Chromomycin A 3, DAPI, Acriflavine-Feulgen reaction, Auramine O-Feulgen reaction, Ethidium Bromide, Propidium iodide, high affinity DNA fluorophores, Green Fluorescent Protein fused to DNA binding protein, ACMA, Quinacrine and Acridine Orange, Feulgen reagent, Gallocyanin chrom-alum, Gallocyanin chrom-alum and naphthol yellow S, Methyl green-pyronin Y and Thionin-Feulgen reagent.
Claim 32
Independent Claim
201 words
A method of in situ analysis of a biological sample comprising the steps of: (a) staining the biological sample with a plurality of 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 a provision that: (i) if said first stain is said first immunohistochemical stain then said second stain is either said second histological stain or said second DNA ploidy stain; (ii) if said first stain is said first histological stain then said second stain is either said second immunohistochemical stain or said second DNA ploidy stain; whereas (iii) if said first stain is said first DNA ploidy stain then said second stain is either said second immunohistochemical stain or said second histological stain; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said plurality of stains are selected such that a spectral component associated with each of said plurality of stains is collectable.
Claim 33
Independent Claim
201 words
A method of in situ analysis of a biological sample comprising the steps of: (a) staining the biological sample with a plurality of 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 a provision that: (i) if said first stain is said first immunohistochemical stain then said second stain is either said second histological stain or said second DNA ploidy stain; (ii) if said first stain is said first histological stain then said second stain is either said second immunohistochemical stain or said second DNA ploidy stain; whereas (iii) if said first stain is said first DNA ploidy stain then said second stain is either said second immunohistochemical stain or said second histological stain; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said plurality of stains are selected such that a spectral component associated with each of said plurality of stains is collectable.
Claim 34
Dependent Claim
37 words
The method of claim 31, wherein said spectral data collection device is selected from the group consisting of an interferometer-based spectral data collection device, filters-based spectral data collection device and a dispersion element-based spectral data collection device.
Claim 35
Dependent Claim
18 words
The method of claim 31, wherein said first and second immunohistochemical stains each independently includes a primary antibody.
Claim 36
Dependent Claim
159 words
The method of claim 33, wherein said primary antibody is selected from the group consisting of anti-estrogen receptor antibody, anti-progesterone receptor antibody, anti-p53 antibody, anti-Her-2/neu antibody, anti-EGFR antibody, anti-cathepsin D antibody, anti-Bcl-2 antibody, anti-E-cadherin antibody, anti-CA125 antibody, anti-CA15-3 antibody, anti-CA19-9 antibody, anti-c-erbB-2 antibody, anti-P-glycoprotein antibody, anti-CEA antibody, anti-retinoblastoma protein antibody, anti-ras oncoprotein antibody, anti-Lewis X antibody, anti-Ki-67 antibody, anti-PCNA antibody, anti-CD3 antibody, anti-CD4 antibody, anti-CD5 antibody, anti-CD7 antibody, anti-CD8 antibody, anti-CD9/p24 antibody, anti-CD10 antibody, anti-CD11c antibody, anti-CD13 antibody, anti-CD14 antibody, anti-CD15 antibody, anti-CD19 antibody, anti-CD20 antibody, anti-CD22 antibody, anti-CD23 antibody, anti-CD30 antibody, anti-CD31 antibody, anti-CD33 antibody, anti-CD34 antibody, anti-CD35 antibody, anti-CD38 antibody, anti-CD41 antibody, anti-LCA/CD45 antibody, anti-CD45RO antibody, anti-CD45RA antibody, anti-CD39 antibody, anti-CD100 antibody, anti-CD95/Fas antibody, anti-CD99 antibody, anti-CD106 antibody, anti-ubiquitin antibody, anti-CD71 antibody, anti-c-myc antibody, anti-cytokeratins antibody, anti-vimentins antibody, anti-HPV proteins antibody, anti-kappa light chains antibody, anti-lambda light chains antibody, anti-melanosomes antibody, anti-prostate specific antigen antibody, anti-S-100 antibody, anti-tau antigen antibody, anti-fibrin antibody, anti-keratins antibody and anti-Tn-antigen antibody.
Claim 37
Dependent Claim
15 words
The method of claim 33, wherein said primary antibody is labeled with a detectable moiety.
Claim 38
Dependent Claim
13 words
The method of claim 35, wherein said detectable moiety is a fluorescent dye.
Claim 39
Dependent Claim
120 words
The method of claim 36, wherein said fluorescent dye is selected from the group consisting of Fluorescein, Rhodamine, Texas Red, Cy2, Cy3, Cy5, VECTOR Red, ELF™ (Enzyme-Labeled Fluorescence), Cy0, Cy0.5, Cy1, Cy1.5, Cy3, Cy3.5, Cy5, Cy7, FluorX, Calcein, Calcein-AM, CRYPTOFLUOR™'S, Orange (42 kDa), Tangerine (35 kDa), Gold (31 kDa), Red (42 kDa), Crimson (40 kDa), BHMP, BHDMAP, Br-Oregon, Lucifer Yellow, Alexa dye family, N-[6-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino]caproyl] (NBD), BODIPY™, boron dipyrromethene difluoride, Oregon Green, MITOTRACKER™ Red, DiOC 7 (3), DiIC 18 , Phycoerythrin, Phycobiliproteins BPE (240 kDa) RPE (240 kDa) CPC (264 kDa) APC (104 kDa), Spectrum Blue, Spectrum Aqua, Spectrum Green, Spectrum Gold, Spectrum Orange, Spectrum Red, NADH, NADPH, FAD, Infra-Red (IR) Dyes, Cyclic GDP-Ribose (cGDPR), Calcofluor White, Tyrosine and Tryptophan.
Claim 40
Dependent Claim
13 words
The method of claim 35, wherein said detectable moiety is a non-fluorescent dye.
Claim 41
Dependent Claim
13 words
The method of claim 38, wherein said non-fluorescent dye includes a heavy metal.
Claim 42
Dependent Claim
26 words
The method of claim 35, wherein said detectable moiety is an enzyme catalyzing a colorimetric reaction of a substrate having a substantially non-soluble color reaction product.
Claim 43
Dependent Claim
23 words
The method of claim 40, wherein said enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase, and glucose oxidase.
Claim 44
Dependent Claim
27 words
The method of claim 40, wherein said substrate is selected from the group consisting of alkaline phosphatase substrates, horseradish peroxidase substrates, glucose oxidase substrates and beta-galactosidase substrates.
Claim 45
Dependent Claim
49 words
The method of claim 35, wherein said detectable moiety is an enzyme catalyzing a luminescence reaction of a substrate having a substantially non-soluble reaction product capable of luminescencing or of chemical involvement in a second reaction with at least one additional substrate, said second reaction having a luminescencing product.
Claim 46
Dependent Claim
23 words
The method of claim 43, wherein said enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase and glucose oxidase.
Claim 47
Dependent Claim
32 words
The method of claim 43, wherein said first and at least one additional substrates are each independently selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase and glucose oxidase substrates.
Claim 48
Dependent Claim
23 words
The method of claim 31, wherein said first and second immunohistochemical stains each independently includes a primary antibody and a signal amplification mechanism.
Claim 49
Dependent Claim
55 words
The method of claim 46, wherein said signal amplification mechanism is selected from the group consisting of a secondary antibody capable of binding a constant region of said primary antibody, avidin or strepavidin capable of binding biotin conjugated to said primary antibody and biotin capable of binding avidin or strepavidin conjugated to said primary antibody.
Claim 50
Dependent Claim
21 words
The method of claim 44, wherein said secondary antibody, avidin, strepavidin and biotin are each independently labeled with a detectable moiety.
Claim 51
Dependent Claim
13 words
The method of claim 48, wherein said detectable moiety is a fluorescent dye.
Claim 52
Dependent Claim
120 words
The method of claim 49, wherein said fluorescent dye is selected from the group consisting of Fluorescein, Rhodamine, Texas Red, Cy2, Cy3, Cy5, VECTOR Red, ELF™ (Enzyme-Labeled Fluorescence), Cy0, Cy0.5, Cy1, Cy1.5, Cy3, Cy3.5, Cy5, Cy7, FluorX, Calcein, Calcein-AM, CRYPTOFLUOR™'S, Orange (42 kDa), Tangerine (35 kDa), Gold (31 kDa), Red (42 kDa), Crimson (40 kDa), BHMP, BHDMAP, Br-Oregon, Lucifer Yellow, Alexa dye family, N-[6-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino]caproyl] (NBD), BODIPY™, boron dipyrromethene difluoride, Oregon Green, MITOTRACKER™ Red, DiOC 7 (3), DiIC 18 , Phycoerythrin, Phycobiliproteins BPE (240 kDa) RPE (240 kDa) CPC (264 kDa) APC (104 kDa), Spectrum Blue, Spectrum Aqua, Spectrum Green, Spectrum Gold, Spectrum Orange, Spectrum Red, NADH, NADPH, FAD, Infra-Red (IR) Dyes, Cyclic GDP-Ribose (cGDPR), Calcofluor White, Tyrosine and Tryptophan.
Claim 53
Dependent Claim
13 words
The method of claim 48, wherein said detectable moiety is a non-fluorescent dye.
Claim 54
Dependent Claim
25 words
The method of claim 51, wherein said non-fluorescent dye is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, glucose oxidase and beta-galactosidase substrates.
Claim 55
Dependent Claim
26 words
The method of claim 48, wherein said detectable moiety is an enzyme catalyzing a colorimetric reaction of a substrate having a substantially non-soluble color reaction product.
Claim 56
Dependent Claim
23 words
The method of claim 53, wherein said enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase, and glucose oxidase.
Claim 57
Dependent Claim
24 words
The method of claim 53, wherein said substrate is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, β-galactosidase, and glucose oxidase substrates.
Claim 58
Dependent Claim
42 words
The method of claim 48, wherein said detectable moiety is an enzyme catalyzing a luminescence reaction of a substrate having a substantially non-soluble reaction product capable of luminescencing or of directing a second reaction of a second substrate having a luminescencing product.
Claim 59
Dependent Claim
18 words
The method of claim 56, wherein said enzyme is selected from the group consisting of luciferase and aequorin.
Claim 60
Dependent Claim
27 words
The method of claim 56, wherein said first and second substrates are each independently selected from the group consisting of luciferine, ATP, coelenterazine and Ca ++ .
Claim 61
Dependent Claim
66 words
The method of claim 31, wherein said first and second histological stains are each independently selected from the group consisting of 4',6-diamidino-2-phenylindole, Eosin, Fluorescein isothiocyanate, Hoechst 33258, Hoechst 33342, Propidium Iodide, Quinacrine, Fluorescein-phalloidin, Resorufin, hematoxylin, Orange G, Light Green SF, Romanowsky-Giemsa, May-Grunwald, Blue counterstain, ethyl green, Feulgen-naphthol yellow S, Giemsa, Methylene Blue, Methyl Green, pyronin, Naphthol-yellow, Neutral Red, Papanicolaou stain, Red Counterstain C and Sirius Red.
Claim 62
Independent Claim
73 words
A method of in situ analysis of a biological sample comprising the steps of: (a) staining the biological sample with at least four different immunohistochemical stains; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said at least four immunohistochemical stains are selected such that a spectral component associated with each of said at least four immunohistochemical stains is collectable.
Claim 63
Independent Claim
73 words
A method of in situ analysis of a biological sample comprising the steps of: (a) staining the biological sample with at least four different immunohistochemical stains; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said at least four immunohistochemical stains are selected such that a spectral component associated with each of said at least four immunohistochemical stains is collectable.
Claim 64
Independent Claim
94 words
The method of in situ analysis of a biological sample comprising the steps of: (a) staining the biological sample with at least three stains of which at least one stain is an immunohistochemical stain and at least one additional stain is a histological stain or a DNA ploidy stain; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said at least three stains are selected such that a spectral component associated with each of said at least three stains is collectable.
Claim 65
Independent Claim
94 words
The method of in situ analysis of a biological sample comprising the steps of: (a) staining the biological sample with at least three stains of which at least one stain is an immunohistochemical stain and at least one additional stain is a histological stain or a DNA ploidy stain; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said at least three stains are selected such that a spectral component associated with each of said at least three stains is collectable.
Claim 66
Independent Claim
95 words
A method of in situ analysis of a biological sample comprising the steps of: (a) staining the biological sample with at least three stains of which a first stain is an immunohistochemical stain, a second stain is a histological stain and a third stain is a DNA ploidy stain; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said at least three stains are selected such that a spectral component specifically associated with each of said at least three stains is collectable.
Claim 67
Independent Claim
95 words
A method of in situ analysis of a biological sample comprising the steps of: (a) staining the biological sample with at least three stains of which a first stain is an immunohistochemical stain, a second stain is a histological stain and a third stain is a DNA ploidy stain; and (b) using a spectral data collection device for collecting spectral data from said biological sample, said spectral data collection device and said at least three stains are selected such that a spectral component specifically associated with each of said at least three stains is collectable.
Claim 68
Dependent Claim
35 words
An immunohistochemical composition comprising at least four different immunohistochemical stains, each being for staining a respective cytological marker and each being individually detectable in a presence of all others using a spectral data collection device.
Claim 69
Dependent Claim
35 words
An immunohistochemical composition comprising at least four different immunohistochemical stains, each being for staining a respective cytological marker and each being individually detectable in a presence of all others using a spectral data collection device.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2002-12-27 | 2003-06-28 | 2003-12-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2006-12-27 | 2007-06-28 | 2007-12-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2010-12-27 | 2011-06-28 | 2011-12-28 | $7700.00 | Window Closed / Lapsed |