To characterize a new protocol of computed tomographic (CT) fluoroscopy-guided biopsy of the lung and upper abdomen to minimize the intervention time, complication rate, and exposure to ionizing radiation for both the patient and the radiologist. Fifty patients (23 women, 27 men; mean age, 64.3 years; age range, 36-83 years) with lung (n = 41) or upper abdomen (n = 9) nodules 15 mm or smaller underwent CT fluoroscopy-guided biopsy from November 2005 to October 2006. The mean nodule diameter was 12.6 mm (range, 8-15 mm), the mean depth to skin was 57.3 mm (range, 20-114 mm), and the mean depth of nodules from pleura and/or peritoneum was 18.9 mm (range, 1-77 mm). Histopathologic evaluation of samples was performed on the day of the procedure. A CT fluoroscopy-guided biopsy protocol was established as follows: (a) native CT with breath-holding at an intermediate respiration level, (b) selection of section position with target nodule and insertion of an 18-gauge coaxial biopsy needle extrapleurally and/or extraperitoneally virtually targeting at nodule, (c) start of CT fluoroscopy (130 kVp, 30 mAs, 5-mm-thick sections) at inspiration level with the patient expiring, (d) stop of CT fluoroscopy when the target nodule reaches the section position, short breath-hold, needle advancement to the target nodule, (e) control of needle position with CT fluoroscopy, and (f) biopsy. The mean total table time was 23.8 minutes (range, 15-41 minutes), the mean duration of CT fluoroscopy was 8.2 seconds (range, 4-23 seconds), and the mean duration of breath-holding--including needle insertion to target nodule and control CT fluoroscopy--was 10.3 seconds (range, 5-15 seconds). There were three minor pneumothoraces that required no further intervention, seven minor pulmonary hemorrhages, three moderate pulmonary hemorrhages with hemoptysis, and one moderate liver hematoma. There were no major complications. The diagnostic accuracy of biopsy samples was 96%. The presented modification of CT fluoroscopy-guided biopsy of mobile pulmonary and upper abdominal lesions is a rapid and safe procedure, requiring only short exposure to ionizing radiation.