Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies
In the development of a multiplex immunofluorescence (IF) platform and the optimization and validation of new multiplex IF panels using a tyramide signal amplification system, several technical requirements are important for high-quality staining, analysis, and results. The aim of this review is to...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-01-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/12/2/255 |
| _version_ | 1797725972359806976 |
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| author | Edwin Roger Parra Mei Jiang Luisa Solis Barbara Mino Caddie Laberiano Sharia Hernandez Swati Gite Anuj Verma Michael Tetzlaff Cara Haymaker Auriole Tamegnon Jaime Rodriguez-Canales Clifford Hoyd Chantale Bernachez Ignacio Wistuba |
| author_facet | Edwin Roger Parra Mei Jiang Luisa Solis Barbara Mino Caddie Laberiano Sharia Hernandez Swati Gite Anuj Verma Michael Tetzlaff Cara Haymaker Auriole Tamegnon Jaime Rodriguez-Canales Clifford Hoyd Chantale Bernachez Ignacio Wistuba |
| author_sort | Edwin Roger Parra |
| collection | DOAJ |
| description | In the development of a multiplex immunofluorescence (IF) platform and the optimization and validation of new multiplex IF panels using a tyramide signal amplification system, several technical requirements are important for high-quality staining, analysis, and results. The aim of this review is to discuss the basic requirements for performing multiplex IF tyramide signal amplification (TSA) in formalin-fixed, paraffin-embedded cancer tissues to support translational oncology research. Our laboratory has stained approximately 4000 formalin-fixed, paraffin-embedded tumor samples using the multiplex IF TSA system for immune profiling of several labeled biomarkers in a single slide to elucidate cancer biology at a protein level and identify therapeutic targets and biomarkers. By analyzing several proteins in thousands of cells on a single slide, this technique provides a systems-level view of various processes in various tumor tissues. Although this technology shows high flexibility in cancer studies, it presents several challenges when applied to study different histology cancers. Our experience shows that adequate antibody validation, staining optimization, analysis strategies, and data generation are important steps for generating quality results. Tissue management, fixation procedures, storage, and cutting can also affect the results of the assay and must be standardized. Overall, this method is reliable for supporting translational research given a precise, step-by-step approach. |
| first_indexed | 2024-03-12T10:39:00Z |
| format | Article |
| id | doaj.art-c0becc0c735943f39fd1e299e8621c83 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-12T10:39:00Z |
| publishDate | 2020-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-c0becc0c735943f39fd1e299e8621c832023-09-02T08:23:41ZengMDPI AGCancers2072-66942020-01-0112225510.3390/cancers12020255cancers12020255Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology StudiesEdwin Roger Parra0Mei Jiang1Luisa Solis2Barbara Mino3Caddie Laberiano4Sharia Hernandez5Swati Gite6Anuj Verma7Michael Tetzlaff8Cara Haymaker9Auriole Tamegnon10Jaime Rodriguez-Canales11Clifford Hoyd12Chantale Bernachez13Ignacio Wistuba14Department of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Pathology, Division of Pathology and Laboratory Medicine. The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USATranslational Medicine, Oncology Research & Development, AstraZeneca, Gaithersburg, MD 20878, USAAkoya Biosciences, Hopkinton, MA 01748, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Translational Molecular Pathology, the University of Texas MD Anderson Cancer Center, Houston, TX 77030, USAIn the development of a multiplex immunofluorescence (IF) platform and the optimization and validation of new multiplex IF panels using a tyramide signal amplification system, several technical requirements are important for high-quality staining, analysis, and results. The aim of this review is to discuss the basic requirements for performing multiplex IF tyramide signal amplification (TSA) in formalin-fixed, paraffin-embedded cancer tissues to support translational oncology research. Our laboratory has stained approximately 4000 formalin-fixed, paraffin-embedded tumor samples using the multiplex IF TSA system for immune profiling of several labeled biomarkers in a single slide to elucidate cancer biology at a protein level and identify therapeutic targets and biomarkers. By analyzing several proteins in thousands of cells on a single slide, this technique provides a systems-level view of various processes in various tumor tissues. Although this technology shows high flexibility in cancer studies, it presents several challenges when applied to study different histology cancers. Our experience shows that adequate antibody validation, staining optimization, analysis strategies, and data generation are important steps for generating quality results. Tissue management, fixation procedures, storage, and cutting can also affect the results of the assay and must be standardized. Overall, this method is reliable for supporting translational research given a precise, step-by-step approach.https://www.mdpi.com/2072-6694/12/2/255tumor immune profilingmultiplex immunofluorescencetyramide signal amplificationimage analysis |
| spellingShingle | Edwin Roger Parra Mei Jiang Luisa Solis Barbara Mino Caddie Laberiano Sharia Hernandez Swati Gite Anuj Verma Michael Tetzlaff Cara Haymaker Auriole Tamegnon Jaime Rodriguez-Canales Clifford Hoyd Chantale Bernachez Ignacio Wistuba Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies Cancers tumor immune profiling multiplex immunofluorescence tyramide signal amplification image analysis |
| title | Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies |
| title_full | Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies |
| title_fullStr | Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies |
| title_full_unstemmed | Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies |
| title_short | Procedural Requirements and Recommendations for Multiplex Immunofluorescence Tyramide Signal Amplification Assays to Support Translational Oncology Studies |
| title_sort | procedural requirements and recommendations for multiplex immunofluorescence tyramide signal amplification assays to support translational oncology studies |
| topic | tumor immune profiling multiplex immunofluorescence tyramide signal amplification image analysis |
| url | https://www.mdpi.com/2072-6694/12/2/255 |
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