Optimized double immunofluorescence in the pathologic diagnosis of non-small cell lung cancer
ZHANG Xian
XU Jiao
SUN Hui-yuan
Abstract：Objective: To investigate the application value of optimized dual immunofluorescence in evaluating the expression of biological molecules in the tumor microenvironment (TME) of non-small cell lung cancer (NSCLC). Methods: A total of 50 cases of histologically confirmed NSCLC tissue blocks were selected from Wujin People's Hospital of Changzhou between March 2020 and June 2023. Each case had 10 sections. Three sections were used for single-label immunofluorescence staining with IL-33 (goat anti), IL-33 (mouse anti), and E-cad (mouse anti), and three sections were used as negative controls. The remaining four sections were divided into two groups based on the species of the primary antibody: same-species sources: experimental groups I and II, with group I as the same-species control; cross-species sources: experimental groups III and IV, with group III as the cross-species control, for dual immunofluorescence staining. The staining quality was evaluated by senior pathologists. Results: After optimization, the dual immunofluorescence staining showed moderate fluorescence intensity, clean background, and high signal-to-noise ratio, enabling qualitative, semi-quantitative, and co-localization analysis. Conclusion: This optimized protocol solved the difficulties in selecting primary antibody species for dual immunofluorescence technology and significantly reduced non-specific staining, providing feasibility for its widespread application. It can be promoted in clinical research and pathological diagnosis.
Keywords：Dual Immunofluorescence TechnologyTumor MicroenvironmentNon-Small Cell Lung CancerEpithelial-Mesenchymal Transition
Publication Date：2025-02-27
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 218-222 )