Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping
Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still pre...
Ausführliche Beschreibung
Autor*in: |
Tubbs, Raymond R. [verfasserIn] |
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E-Artikel |
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Sprache: |
Englisch |
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2007 |
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Anmerkung: |
© Springer Science+Business Media, Inc. 2007 |
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Übergeordnetes Werk: |
Enthalten in: The histochemical journal - Dordrecht [u.a.] : Springer Science + Business Media B.V., 1968, 38(2007), 2 vom: 05. Jan., Seite 129-134 |
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Übergeordnetes Werk: |
volume:38 ; year:2007 ; number:2 ; day:05 ; month:01 ; pages:129-134 |
Links: |
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DOI / URN: |
10.1007/s10735-006-9074-1 |
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520 | |a Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. | ||
650 | 4 | |a Tyramide signal amplification |7 (dpeaa)DE-He213 | |
650 | 4 | |a Catalyzed reporter deposition |7 (dpeaa)DE-He213 | |
650 | 4 | |a Fluorescence in situ hybridization |7 (dpeaa)DE-He213 | |
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650 | 4 | |a Glioblastoma multiforme |7 (dpeaa)DE-He213 | |
700 | 1 | |a Das, Kingshuk |4 aut | |
700 | 1 | |a Cook, James R. |4 aut | |
700 | 1 | |a Pettay, James D. |4 aut | |
700 | 1 | |a Roche, Patrick C. |4 aut | |
700 | 1 | |a Grogan, Thomas |4 aut | |
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10.1007/s10735-006-9074-1 doi (DE-627)SPR012830070 (SPR)s10735-006-9074-1-e DE-627 ger DE-627 rakwb eng Tubbs, Raymond R. verfasserin aut Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping 2007 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer Science+Business Media, Inc. 2007 Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. Tyramide signal amplification (dpeaa)DE-He213 Catalyzed reporter deposition (dpeaa)DE-He213 Fluorescence in situ hybridization (dpeaa)DE-He213 Glioma (dpeaa)DE-He213 Glioblastoma multiforme (dpeaa)DE-He213 Das, Kingshuk aut Cook, James R. aut Pettay, James D. aut Roche, Patrick C. aut Grogan, Thomas aut Enthalten in The histochemical journal Dordrecht [u.a.] : Springer Science + Business Media B.V., 1968 38(2007), 2 vom: 05. Jan., Seite 129-134 (DE-627)503327638 (DE-600)2210318-1 1573-6865 nnns volume:38 year:2007 number:2 day:05 month:01 pages:129-134 https://dx.doi.org/10.1007/s10735-006-9074-1 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_120 GBV_ILN_138 GBV_ILN_152 GBV_ILN_161 GBV_ILN_171 GBV_ILN_187 GBV_ILN_224 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 AR 38 2007 2 05 01 129-134 |
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10.1007/s10735-006-9074-1 doi (DE-627)SPR012830070 (SPR)s10735-006-9074-1-e DE-627 ger DE-627 rakwb eng Tubbs, Raymond R. verfasserin aut Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping 2007 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer Science+Business Media, Inc. 2007 Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. Tyramide signal amplification (dpeaa)DE-He213 Catalyzed reporter deposition (dpeaa)DE-He213 Fluorescence in situ hybridization (dpeaa)DE-He213 Glioma (dpeaa)DE-He213 Glioblastoma multiforme (dpeaa)DE-He213 Das, Kingshuk aut Cook, James R. aut Pettay, James D. aut Roche, Patrick C. aut Grogan, Thomas aut Enthalten in The histochemical journal Dordrecht [u.a.] : Springer Science + Business Media B.V., 1968 38(2007), 2 vom: 05. Jan., Seite 129-134 (DE-627)503327638 (DE-600)2210318-1 1573-6865 nnns volume:38 year:2007 number:2 day:05 month:01 pages:129-134 https://dx.doi.org/10.1007/s10735-006-9074-1 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_120 GBV_ILN_138 GBV_ILN_152 GBV_ILN_161 GBV_ILN_171 GBV_ILN_187 GBV_ILN_224 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 AR 38 2007 2 05 01 129-134 |
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10.1007/s10735-006-9074-1 doi (DE-627)SPR012830070 (SPR)s10735-006-9074-1-e DE-627 ger DE-627 rakwb eng Tubbs, Raymond R. verfasserin aut Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping 2007 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer Science+Business Media, Inc. 2007 Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. Tyramide signal amplification (dpeaa)DE-He213 Catalyzed reporter deposition (dpeaa)DE-He213 Fluorescence in situ hybridization (dpeaa)DE-He213 Glioma (dpeaa)DE-He213 Glioblastoma multiforme (dpeaa)DE-He213 Das, Kingshuk aut Cook, James R. aut Pettay, James D. aut Roche, Patrick C. aut Grogan, Thomas aut Enthalten in The histochemical journal Dordrecht [u.a.] : Springer Science + Business Media B.V., 1968 38(2007), 2 vom: 05. Jan., Seite 129-134 (DE-627)503327638 (DE-600)2210318-1 1573-6865 nnns volume:38 year:2007 number:2 day:05 month:01 pages:129-134 https://dx.doi.org/10.1007/s10735-006-9074-1 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_120 GBV_ILN_138 GBV_ILN_152 GBV_ILN_161 GBV_ILN_171 GBV_ILN_187 GBV_ILN_224 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 AR 38 2007 2 05 01 129-134 |
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10.1007/s10735-006-9074-1 doi (DE-627)SPR012830070 (SPR)s10735-006-9074-1-e DE-627 ger DE-627 rakwb eng Tubbs, Raymond R. verfasserin aut Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping 2007 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer Science+Business Media, Inc. 2007 Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. Tyramide signal amplification (dpeaa)DE-He213 Catalyzed reporter deposition (dpeaa)DE-He213 Fluorescence in situ hybridization (dpeaa)DE-He213 Glioma (dpeaa)DE-He213 Glioblastoma multiforme (dpeaa)DE-He213 Das, Kingshuk aut Cook, James R. aut Pettay, James D. aut Roche, Patrick C. aut Grogan, Thomas aut Enthalten in The histochemical journal Dordrecht [u.a.] : Springer Science + Business Media B.V., 1968 38(2007), 2 vom: 05. Jan., Seite 129-134 (DE-627)503327638 (DE-600)2210318-1 1573-6865 nnns volume:38 year:2007 number:2 day:05 month:01 pages:129-134 https://dx.doi.org/10.1007/s10735-006-9074-1 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_120 GBV_ILN_138 GBV_ILN_152 GBV_ILN_161 GBV_ILN_171 GBV_ILN_187 GBV_ILN_224 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 AR 38 2007 2 05 01 129-134 |
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10.1007/s10735-006-9074-1 doi (DE-627)SPR012830070 (SPR)s10735-006-9074-1-e DE-627 ger DE-627 rakwb eng Tubbs, Raymond R. verfasserin aut Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping 2007 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Springer Science+Business Media, Inc. 2007 Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. Tyramide signal amplification (dpeaa)DE-He213 Catalyzed reporter deposition (dpeaa)DE-He213 Fluorescence in situ hybridization (dpeaa)DE-He213 Glioma (dpeaa)DE-He213 Glioblastoma multiforme (dpeaa)DE-He213 Das, Kingshuk aut Cook, James R. aut Pettay, James D. aut Roche, Patrick C. aut Grogan, Thomas aut Enthalten in The histochemical journal Dordrecht [u.a.] : Springer Science + Business Media B.V., 1968 38(2007), 2 vom: 05. Jan., Seite 129-134 (DE-627)503327638 (DE-600)2210318-1 1573-6865 nnns volume:38 year:2007 number:2 day:05 month:01 pages:129-134 https://dx.doi.org/10.1007/s10735-006-9074-1 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_32 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_90 GBV_ILN_95 GBV_ILN_100 GBV_ILN_101 GBV_ILN_105 GBV_ILN_120 GBV_ILN_138 GBV_ILN_152 GBV_ILN_161 GBV_ILN_171 GBV_ILN_187 GBV_ILN_224 GBV_ILN_250 GBV_ILN_267 GBV_ILN_281 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_702 AR 38 2007 2 05 01 129-134 |
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Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping Tyramide signal amplification (dpeaa)DE-He213 Catalyzed reporter deposition (dpeaa)DE-He213 Fluorescence in situ hybridization (dpeaa)DE-He213 Glioma (dpeaa)DE-He213 Glioblastoma multiforme (dpeaa)DE-He213 |
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Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping |
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Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. © Springer Science+Business Media, Inc. 2007 |
abstractGer |
Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. © Springer Science+Business Media, Inc. 2007 |
abstract_unstemmed |
Abstract Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically—defined cell populations for gated genotyping. © Springer Science+Business Media, Inc. 2007 |
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The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. 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7.39824 |