Quantitative Immunohistochemistry of Tumor Immune Cells

Tumors were formalin-fixed and paraffin-embedded and processed into 4 μm tissue sections. Tumor sections were stained for FAP (EPR20021, Abcam), CD8 (D4W2Z, Cell Signaling), and CD4 (EPR19514, Abcam) on a Bond Rx autostainer (Leica Biosystems) with Bond Polymer Refine Detection (Leica Biosystems) according to the manufacturer's protocol. After staining, sections were dehydrated and film cover slipped using a TissueTek-Prisma and Coverslipper (Sakura). Whole slide scanning (40x) was performed on an Aperio AT2 (Leica Biosystems). Image analysis and quantification was performed on Aperio ImageScope v12.1 (Leica Biosystems) or QuPath v0.1.2 [18 (link)].

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Zboralski D., Osterkamp F., Christensen E., Bredenbeck A., Schumann A., Hoehne A., Schneider E., Paschke M., Ungewiss J., Haase C., Robillard L., Simmons A.D., Harding T.C, & Nguyen M. (2023). Fibroblast activation protein targeted radiotherapy induces an immunogenic tumor microenvironment and enhances the efficacy of PD-1 immune checkpoint inhibition. European Journal of Nuclear Medicine and Molecular Imaging, 50(9), 2621-2635.

Publication 2023

D4W2Z EPR19514 Bond Rx Bond Polymer Refine Detection Aperio AT2

Formalin Paraffin Polymer Prisma Tissue Tumor

Corresponding Organization :

Other organizations : Clovis Oncology (United States)

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Variable analysis

independent variables

Tumor sections staining for FAP (EPR20021, Abcam), CD8 (D4W2Z, Cell Signaling), and CD4 (EPR19514, Abcam)

dependent variables

Quantification of FAP, CD8, and CD4 expression in tumor sections

control variables

Formalin-fixed and paraffin-embedded tumor samples
Tissue sections processed into 4 μm thickness
Staining performed on a Bond Rx autostainer (Leica Biosystems) with Bond Polymer Refine Detection (Leica Biosystems) according to the manufacturer's protocol
Dehydration and film cover slipping using a TissueTek-Prisma and Coverslipper (Sakura)
Whole slide scanning (40x) performed on an Aperio AT2 (Leica Biosystems)
Image analysis and quantification performed on Aperio ImageScope v12.1 (Leica Biosystems) or QuPath v0.1.2

controls

No positive or negative controls were explicitly mentioned in the provided information.

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