In this paper, we report an advanced sample preparation methodology using in-situ lift-out FIB and Flipstage for tridirectional TEM failure analysis. A planar-view and two cross-section TEM samples were prepared from the same target. Firstly, a planar-view lamellar parallel to the wafer surface was prepared using in-situ lift-out FIB milling. Upon TEM analysis, the planar sample was further milled in the along-gate and cross-gate directions separately. Eventually, a pillar-like sample containing a single transistor gate was obtained. Using this technique, we are able to analyze the defect from three perpendicular directions and obtain more information on the defect for failure root-cause analysis. A MOSFETs case study is described to demonstrate the procedure and advantages of this technique.