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This study evaluates the feasibility of Electrical Impedance Spectroscopy (EIS) as a minimally invasive technique to differentiate healthy lung tissue from peripheral pulmonary nodules and masses during Electromagnetic Navigation Bronchoscopy (ENB). The diagnostic capability of EIS is also assessed in distinguishing malignant from benign lesions. Significant differences (P < 0.001) were observed between healthy tissue and lesions in all impedance parameters (resistance, reactance, modulus, and phase angle). Differences between benign and malignant nodules and masses were observed in the complex plane plot, but some overlap between both occurred possibly due to similarities between adenocarcinomas and inflammatory nodules, primarily in terms of water content. The findings suggest that EIS could be implemented as a potential tool to improve biopsy guidance, reducing the need for additional X-ray imaging during the procedure. In addition, increasing sample size is needed to enhance diagnostic accuracy.Clinical RelevanceElectrical Impedance Spectroscopy (EIS) has proven to be an effective tool for distinguishing healthy lung tissue from peripheral lung nodules and masses. Using this technique for guiding the biopsy localization could lead to an improved diagnostic yield while reducing dependence on ionizing radiation-based imaging, such as X-rays.