The invention discloses a puncturing positioning device of operation of intervertebral foramen of lumbar vertebra and a using method thereof, and belongs to the field of medical instruments. The operation puncturing positioning device comprises an operation bed fixing clamp, an upright column which is perpendicular to a bed surface is connected with the fixing clamp, the end of a cross bar is slidably connected with the upright column, the other end of the cross bar is a rotary shaft, an adjusting rotary tube is rotatably connected with the rotary shaft, a positioning tube which is perpendicular to the axial center of the rotary shaft is arranged on the side, close the end, of the adjusting rotary tube, a positioning needle is arranged in the positioning tube, a horizontal length measuringbar and a tilting length measuring bar are arranged on a rotary shaft and an adjusting rotary tube which are located on the side of the positioning tube, arc-shaped long holes which are formed in therotary direction are formed in the outer circumference of the adjusting rotary tube, and the arc-shaped long holes move along the horizontal length measuring bar when the adjusting rotary tube is rotated. By means of the puncturing positioning device, the puncturing direction and angle can be accurately positioned, the operation time can be shortened, the smooth processing of the operation is ensured, the pain of a patient is relieved, and the overstrain of a doctor is reduced.