Sections of formalin-fixed paraffin-embedded tissue (10 μm for xenografts and 5μm for brain tissue) were stained with H&E or with an anti-RFP rabbit polyclonal antibody that crossreacts with mStrawberry (1:100 dilution) (Abcam ab34771) and detected using Leica’s Polymer Refine Kit in combination with the Bond automated system (Leica Biosystems DS9800). Slides were scanned at x20 magnification with a resolution of 0.5 μm per pixel on an Aperio AT2 (Leica Biosystems). Images were analyzed using the Aperio Imagescope Nuclear version 9 algorithm to quantify the proportion of cells with positive nuclear staining.
Polymer refine kit
Manufactured by Leica Biosystems
Sourced in United Kingdom
The Polymer Refine Kit is a laboratory equipment product offered by Leica Biosystems. The kit provides a solution for staining and visualizing biological samples. The core function of the product is to facilitate the detection and analysis of target proteins or other molecular markers within tissue samples.
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Lab products found in correlation
3 protocols using polymer refine kit
1
Quantifying Cells in Tissue Samples
2
Quantifying Cells in Tissue Samples
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Sections of formalin-fixed paraffin-embedded tissue (10 μm for xenografts and 5μm for brain tissue) were stained with H&E or with an anti-RFP rabbit polyclonal antibody that crossreacts with mStrawberry (1:100 dilution) (Abcam ab34771) and detected using Leica’s Polymer Refine Kit in combination with the Bond automated system (Leica Biosystems DS9800). Slides were scanned at x20 magnification with a resolution of 0.5 μm per pixel on an Aperio AT2 (Leica Biosystems). Images were analyzed using the Aperio Imagescope Nuclear version 9 algorithm to quantify the proportion of cells with positive nuclear staining.
3
Immunohistochemical Evaluation of p16 in OPSCC
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Immunohistochemical expression of p16 was evaluated in all cases. In brief, deparaffinized FFPE sections of all cases were subjected to antigen retrieval using the Leica Bond protocol (Leica Biosystems) with proprietary Retrieval ER2 (ethylene diamine tetraacetic acid, pH 9.0) buffer for 20 min. A mouse monoclonal antibody against p16 (E6H4 clone, CINtec; Ventana Medical Systems, Arizona, USA) was utilized with a 1:4 dilution and detected with the Polymer Refine Kit (Leica Biosystems, New Castle, UK) on a Leica Bond autostainer. For positive immunohistochemical controls, a tonsil SCC with positive p16 expression was used. The threshold for p16 positivity was met in cases where ≥ 70% of tumor cells demonstrated strong diffuse nuclear and cytoplasmic staining. The threshold of ≥ 70% was used as it correlates with a positive HPV status in OPSCC [20 (link)]. Cases with moderate staining intensity in ≥ 70% of the cells or strong staining reactivity in ≥ 50 < 70% of the cells were categorized as equivocal.
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