Fourier transform infrared (FTIR) spectroscopy with an attenuated total reflection(ATR)probe was used to study the benign and malignant tissues of gallbladder. FTIR spectra of 18 tissues of gallbladder carcinoma and 139 benign tissues were collected and analyzed. The results demonstrated that bands related to lipid, carbohydrate, and nucleic acid from malignant samples were significantly different from those of benign ones: (1) The peak positions of 1 167 and 1 123 cm-1 shifted toward lower wave number (P＜0.05) but that of 1 309 cm-1 to the higher one (P＜0.05); (2) The relative intensity ratios of I2 856/I1 461, I1 167/I1 461, I1 123/I1 461, I1 082/I1 461 increased significantly (P＜0.05) (3)The half-widths of 1 167 and 1 082 cm-1increased significantly (P＜0.05), whereas the half-width of 1 461 cm-1 decreased significantly (P＜0.05). (4) The probability of 1 750 cm-1 band of gallbladder carcinoma tissues increased significantly (P＜0.05).