TY - JOUR
T1 - Best Practices for Technical Reproducibility Assessment of Multiplex Immunofluorescence
AU - Laberiano-Fernández, Caddie
AU - Hernández-Ruiz, Sharia
AU - Rojas, Frank
AU - Parra, Edwin Roger
N1 - Funding Information:
This study was supported in part by the scientific and financial support for the CIMAC-CIDC Network provided through the National Cancer Institute (NCI) Cooperative Agreement U24CA224285 of The University of Texas MD Anderson Cancer Center CIMAC and for the Translational Molecular Pathology Immunoprofiling Platform, as well as by National Institutes of Health/NCI through Cancer Center Support Grant P30CA016672 (Institutional Tissue Bank) and SPORE grant 5P50CA070907-18 from the Cancer Prevention and Research Institute of Texas through MIRA RP160668.
Publisher Copyright:
© Copyright © 2021 Laberiano-Fernández, Hernández-Ruiz, Rojas and Parra.
PY - 2021/8/31
Y1 - 2021/8/31
N2 - Multiplex immunofluorescence (mIF) tyramide signal amplification is a new and useful tool for the study of cancer that combines the staining of multiple markers in a single slide. Several technical requirements are important to performing high-quality staining and analysis and to obtaining high internal and external reproducibility of the results. This review manuscript aimed to describe the mIF panel workflow and discuss the challenges and solutions for ensuring that mIF panels have the highest reproducibility possible. Although this platform has shown high flexibility in cancer studies, it presents several challenges in pre-analytic, analytic, and post-analytic evaluation, as well as with external comparisons. Adequate antibody selection, antibody optimization and validation, panel design, staining optimization and validation, analysis strategies, and correct data generation are important for reproducibility and to minimize or identify possible issues during the mIF staining process that sometimes are not completely under our control, such as the tissue fixation process, storage, and cutting procedures.
AB - Multiplex immunofluorescence (mIF) tyramide signal amplification is a new and useful tool for the study of cancer that combines the staining of multiple markers in a single slide. Several technical requirements are important to performing high-quality staining and analysis and to obtaining high internal and external reproducibility of the results. This review manuscript aimed to describe the mIF panel workflow and discuss the challenges and solutions for ensuring that mIF panels have the highest reproducibility possible. Although this platform has shown high flexibility in cancer studies, it presents several challenges in pre-analytic, analytic, and post-analytic evaluation, as well as with external comparisons. Adequate antibody selection, antibody optimization and validation, panel design, staining optimization and validation, analysis strategies, and correct data generation are important for reproducibility and to minimize or identify possible issues during the mIF staining process that sometimes are not completely under our control, such as the tissue fixation process, storage, and cutting procedures.
KW - analytical evaluation
KW - clinical application
KW - multiplex immunofluorescence
KW - reproducibility
KW - standardization
UR - http://www.scopus.com/inward/record.url?scp=85114891825&partnerID=8YFLogxK
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U2 - 10.3389/fmolb.2021.660202
DO - 10.3389/fmolb.2021.660202
M3 - Review article
C2 - 34532339
AN - SCOPUS:85114891825
SN - 2296-889X
VL - 8
JO - Frontiers in Molecular Biosciences
JF - Frontiers in Molecular Biosciences
M1 - 660202
ER -