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The diagnosis of pulmonary fat embolism (PFE) is of great significance in the field of forensic medicine because it can be considered a major cause of death or a vital reaction. Conventional histological analysis of lung tissue specimens is a widely used method for PFE diagnosis. However, variable and labor-intensive tissue staining procedures impede the validity and informativeness of histological image analysis. To obtain complete information from tissues, a method based on infrared imaging of unlabeled tissue sections was developed to identify pulmonary fat emboli in the present study. We selected 15 PFE-positive lung samples and 15 PFE-negative samples from real cases. Oil red O (ORO) staining and infrared spectral imaging collection were both performed on all lung tissue samples. And the fatty tissue of the abdominal wall and the embolized lipid droplets in the lungs were taken for comparison. The results of the blind, evaluation by pathologists, showed good agreement between the infrared spectral imaging of the lung tissue and the standard histological stained images. Fourier transform infrared (FT-IR) spectroscopic imaging significantly simplifies the typical painstakingly laborious histological staining procedure. And we found a difference between lipid droplets embolized in abdominal wall fat and lung tissue. Graphical Abstract [ABSTRACT FROM AUTHOR]