Of 140 cases of mediastinal neoplasms in our hospital, histological diagnosis was confirmed in 129 cases. We examined the methods of preoperative biopsy with those 129 cases. Biopsy had been performed in 25 cases. Mediastinoscopy was performed in seven cases, needle biopsy in eight cases, lymph node biopsy in eight cases, esophageal biopsy using a gastrofiberscope in one case, transbronchial biopsy using a bronchoscope in one case. The true positive rates of those methods were 100% for both mediastinoscopy and lymph node biopsy, and 75% for needle biopsy. Preoperative misdiagnosis occurred in two cases of needle biopsy. The postoperative histological diagnosis was malignant lymphoma in both cases. We performed gene analysis of the immunoglobulin heavy chain gene, light chain kappa and lambda genes, and the T-cell receptor beta gene by use of biopsied specimens, and we found rearrangement bands of these genes in the cases of malignant lymphoma. Therefore, we summarize that gene analysis is a reliable method if malignant lymphoma is suspected. If a needle biopsy is performed under CT guidance, the needle is sure to puncture the tumor. We concluded, therefore, that if a tumor is located in the anterior mediastinum, CT-guided needle biopsy should be performed first of all. Mediastinoscopy is a useful method if the tumor is located in the mid-mediastinum.