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Background Immunocytochemistry (ICC) is suitable for use on a range of cytology preparations, such as cell blocks, air-dried slides, ethanol-fixed slides, direct smears, cytospins, and liquid-based cytology (LBC) samples. However, it must be standardized against the gold standard of formalin-fixed paraffin-embedded tissues with adequate number of positive and negative controls. The role of ICC in lung cancer is crucial, as most lung cancer specimens are cytology samples. Accurate diagnosis and testing of certain biomarkers rely heavily on both diagnostic and predictive ICC. Summary Key ICC markers important in lung cancer include, but are not limited to, diagnostic ICCs such as TTF-1, p40, Napsin A, and p63, as well as predictive ICCs like ALK, ROS-1, PD-L1, and NTRK. Key Messages With proper validation, immunocytochemistry for lung cancer can be effectively performed on direct smears, cytospins, and other specimens, even when resources for preparing cell blocks are unavailable. This is particularly true for diagnostic antibodies, but it is important to exercise caution with predictive immunocytochemistry. Nonetheless, a low threshold for molecular testing should be maintained. PD-L1 ICC can be challenging and should ideally be performed on formalin-fixed cell blocks or biopsies when available.

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