Echocardiographic diastolic function (DF) assessment remains a challenge in atrial fibrillation (AF), because indexes such as E/A cannot be used and because chronic, rate controlled AF causes chamber remodeling. To determine if echocardiography can accurately characterize diastolic chamber properties we compared 15 chronic AF subjects to 15, age matched normal sinus rhythm (NSR) subjects using simultaneous echocardiography-cardiac catheterization (391 beats analyzed). Conventional DF parameters (DT, E_peak, AT, E_dur, E-VTI, E/E\') and validated, E-wave derived, kinematic modeling based chamber stiffness parameter (k), were compared. For validation, chamber stiffness (dP/dV) was independently determined from simultaneous, multi-beat P-V loop data. Results show that neither AT, E_peak nor E-VTI differentiated between groups. Although DT, E_dur and E/E’ did differentiate between groups (DT_NSR vs. DT_AF p < 0.001, E_durNSR vs. E_durAF p < 0.001, E/E\'_NSR vs. E/E\'_AF p < 0.05), the model derived chamber stiffness parameter k was the only parameter specific for chamber stiffness, (k_NSR vs. k_AF p < 0.005). The invasive gold standard determined end-diastolic stiffness in NSR was indistinguishable from end-diastolic (i.e. diastatic) stiffness in AF (p = 0.84). Importantly, the analysis provided mechanistic insight by showing that diastatic stiffness in AF was significantly greater than diastatic stiffness in NSR (p < 0.05). We conclude that passive (diastatic) chamber stiffness is increased in normal LVEF chronic, rate controlled AF hearts relative to normal LVEF NSR controls and that in addition to DT, the E-wave derived, chamber stiffness specific index k, differentiates between AF vs. NSR groups, even when invasively determined end-diastolic chamber stiffness fails to do so.