Immunohistochemical Scoring of Brain Tumors

Staining was assessed at × 200 magnification following high-resolution scanning (Nanozoomer Digital Pathology Scanner, Hamamatsu Photonics). Assessment was conducted with involvement and training provided by a consultant neuropathologist (SP). Cytoplasmic staining was semi-quantitatively assessed using an immunohistochemical H-score [27 (link)], where staining intensity was assessed as none (0), weak (1), medium (2), or strong (3) over the percentage area of each staining intensity. Nuclear staining was assessed as the percentage of nuclei with any intensity of staining. 30% of cores for each protein were examined by a second independent assessor blinded to clinical outcome and the primary assessor’s scores. Good concordance was demonstrated between scorers (single measure intraclass correlation coefficients were above 0.7 for all markers assessed across all brain tumour cohorts) (Supplementary Table 4). Unbiased cut-points, for stratification, were obtained based on overall survival using X-tile software [28 (link)].

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Yao A., Storr S.J., Al-hadyan K., Rahman R., Smith S., Grundy R., Paine S, & Martin S.G. (2020). Thioredoxin System Protein Expression Is Associated with Poor Clinical Outcome in Adult and Paediatric Gliomas and Medulloblastomas. Molecular Neurobiology, 57(7), 2889-2901.

Publication 2020

Nanozoomer Digital Pathology Scanner

Brain tumour Consultant Cytoplasmic Neuropathologist Nuclei Protein Scanner Weak

Corresponding Organization :

Other organizations : University of Nottingham, Brain Tumour Research, Nottingham University Hospitals NHS Trust, Queen's Medical Centre

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Variable analysis

independent variables

Staining intensity (none, weak, medium, strong)

dependent variables

Cytoplasmic staining (semi-quantitatively assessed using H-score)
Nuclear staining (percentage of nuclei with any intensity of staining)

control variables

Magnification (× 200)
Assessment conducted by a consultant neuropathologist (SP)
30% of cores for each protein were examined by a second independent assessor blinded to clinical outcome and the primary assessor's scores
Good concordance between scorers (single measure intraclass correlation coefficients were above 0.7 for all markers assessed across all brain tumour cohorts)

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