Selecting the appropriate operation for gallbladder cancer depends on the depth of cancer invasion, which remains difficult to determine preoperatively, especially with respect to the subserosal layer (pT2). We devised a laparoscopic cholecystectomy with gallbladder bed dissection (LC with GBD) as a new total biopsy method for suspected gallbladder cancer. We retrospectively reviewed the medical records of 19 patients who underwent LC with GBD to assess the usefulness of this procedure and the pathological findings. No severe morbidity or recurrence was encountered. LC with GBD could be performed easily and safely, and the patients' postoperative course was almost equal to that of patients treated by conventional LC. Histologically, gallbladder cancer was diagnosed in five cases (pT1a, 3; pT2, 2). We believe that LC with GBD could play an important role in the potential treatment strategy for pT2 gallbladder cancer.