Purpose: To evaluate the variability in heart position in deep-inspiration breath-hold (DIBH) radiotherapy for breast cancer when 3D surface imaging would be used for monitoring the depth of the breath hold during treatment. Materials and Methods: Ten patients who received DIBH radiotherapy after breast-conserving surgery (BCS) were included. Retrospectively, heart-based registrations were performed for cone-beam computed tomography (CBCT) to planning CT and breast surface registrations were performed for a 3D surface (two different regions of interest [ROIs]), captured concurrently with CBCT, to planning CT. The resulting setup errors were compared with linear regression analysis and receiver operating characteristic (ROC) analysis was performed to investigate the prediction quality of 3D surface imaging for 3D heart displacement. Further, the residual setup errors (systematic [Σ] and random [σ]) of the heart were estimated relative to the surface registrations. Results: When surface imaging [ROIleft-side;ROIboth-sides] would be used for monitoring, the residual errors of the heart position are in left-right: Σ=[0.36;0.12], σ=[0.16;0.14]; cranio-caudal: Σ=[0.54;0.54], σ=[0.28;0.31]; and in anteriorposterior: Σ=[0.18;0.14], σ=[0.20;0.19] cm. Correlations between setup errors were: R2 = [0.23;0.73], [0.67;0.65], [0.65;0.73] in left-right, cranio-caudal, and anterior-posterior direction, respectively. ROC analysis resulted in an area under the ROC curve of [0.82;0.78]. Conclusion: The use of ROIboth-sides provided promising results. However, considerable variability in the heart position, particularly in CC direction, is observed when 3D surface imaging would be used for guidance in DIBH radiotherapy after BCS. Planning organ at risk volume margins should be used to take into account the heart-position variability.