IHC analysis was performed to assess the expression level of PF4 (ab303494; Abcam) in clear cell renal cell carcinoma (ccRCC) samples from Fudan University Shanghai Cancer Center (FUSCC) following manufacturers’ protocols as previously described [46 (link)]. The mIHC staining assay was conducted to investigate the abundance and distribution of PF4 (ab303494; Abcam), CD68 (ab125212; Abcam), and CD163 (25121; CST) in ccRCC adjacent normal kidney tissues following manufacturers’ protocols. Tissue slides that were bound with primary and secondary antibodies but not fluorophores were included as negative controls to assess autofluorescence. Multiplex stained slides were scanned using a Vectra Polaris Quantitative Pathology Imaging System (Akoya Biosciences) at 20 nm wavelength intervals from 440 to 780 nm with a fixed exposure time and an absolute magnification of 200×. All scans for each slide were then superimposed to obtain a single image. Multilayer images were imported to inForm v.2.4.8 (Akoya Biosciences) for quantitative image analysis.
Xu W., Ye S., Liu W., Guo H., Zhang L., Wei S., Anwaier A., Chang K., Malafaia G., Zhang H., Ye D, & Wei G. (2024). Single-cell RNA-seq analysis decodes the kidney microenvironment induced by polystyrene microplastics in mice receiving a high-fat diet. Journal of Nanobiotechnology, 22, 13.
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