Fig. 2

High resolution image of non-small cell lung cancer PD-L1 expression from formalin-fixed, paraffin-embedded specimens generated using immunofluorescence staining and tissue clearing. A, B The standard H&E staining and PD-L1 immunohistochemistry (IHC) performed on 4 μm sections of one representative case, respectively. The PD-L1 immunofluorescence and tissue clearing protocols produced image (C) was comparable to the traditional IHC, including the membranous staining pattern of PD-L1. The fluorescence image can be converted to pseudo-H&E (D) and pseudo-IHC (E) formats for assessment by pathologists. The three-dimensional projection of the immunofluorescence image (F) shows good tissue details of the entire specimen (Green: PD-L1, red: DiD, and blue: SYTO-16. Scale bar: A–E: 50 μm, F: 150 μm)