With the advent of catheter ablation techniques, precise localization of accessory AV pathways (AP) assumes greater importance. In an effort to define the course of AP fibers, we attempted to record activation of 56 left free-wall and 23 posteroseptal APs in 62 patients undergoing electrophysiological study. The coronary sinus (CS) and great cardiac vein (GCV) were mapped using orthogonal catheter electrodes, which provide a recording dipole perpendicular to the AV groove. The tricuspid annulus (TA) was mapped using a 2 mm spaced octapolar electrode catheter. Potentials were considered to represent AP activation only if they could be dissociated from both atrial and ventricular activation by programmed stimulation. Orthogonal catheter electrodes in the CS and GCV were advanced beyond the site of earliest retrograde atrial activation and/or earliest antegrade ventricular activation in 45 of the 56 left free-wall APs, and AP potentials were recorded from 42 (93%). An oblique course was identified in 36 APs, with the ventricular insertion being recorded 4-30 mm (median 15 mm) distal or anterior to the atrial insertion. In three patients, antegrade and retrograde conduction proceeded over different (but close) parallel fibers. AP potentials were recorded from 19 of 23 posteroseptal pathways. Ten pathways (left posteroseptal) were recorded from the CS, beginning 5-11 mm (median 9 mm) distal to the os, with potentials extending 8-18 mm (median 11 mm) distally. Four pathways (mid-septal) were recorded along the TA, anterior to the CS ostium and posterior to the His bundle catheter. Five pathways (right posteroseptal) were recorded along the TA, directly opposite or immediately posterior to the CS ostium. One of the patients had both midseptal and left posteroseptal pathways and three patients had both right posteroseptal and left posteroseptal pathways. We conclude: 1) left free-wall APs transit the AV groove obliquely and may be comprised of multiple, closely spaced, parallel fibers; 2) the anatomical location of "posteroseptal" pathways is variable and the presence of fibers at multiple sites is common; and 3) direct recordings of AP activation facilitate tracking of the accessory pathway along its course from atrium to ventricle and help identify the presence of multiple fibers.