After the pathologist reached a diagnosis, the number of tumor cells was counted manually by one cytoscreener and one pulmonologist in a blinded manner using HE staining slides. Then, the average number of tumor cells was calculated.
After sectioning of samples to 4–5 μm, PD-L1 staining was performed with 22C3 antibodies (rabbit monoclonal, clone 22C3; Agilent Dako, Glostrup, Denmark) using autostainer (Autostainer Link 48, Agilent Dako). PD-L1 positivity was defined as membranous staining in at least 1% of cells [10 (link)], regardless of staining intensity and proportion in the membrane. PD-L1 was evaluated by experienced pathologist, and the cut off values were classified as ≥50% and ≥ 1%. The number of tumor cells by a single biopsy, total number of tumor cells, average number of tumor cells, and PD-L1 expression for each patient were compared between Cryo and TBB.
After sectioning of samples to 4–5 μm, PD-L1 staining was performed with 22C3 antibodies (rabbit monoclonal, clone 22C3; Agilent Dako, Glostrup, Denmark) using autostainer (Autostainer Link 48, Agilent Dako). PD-L1 positivity was defined as membranous staining in at least 1% of cells [10 (link)], regardless of staining intensity and proportion in the membrane. PD-L1 was evaluated by experienced pathologist, and the cut off values were classified as ≥50% and ≥ 1%. The number of tumor cells by a single biopsy, total number of tumor cells, average number of tumor cells, and PD-L1 expression for each patient were compared between Cryo and TBB.
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