Accelerated partial breast irradiation is an attractive alternative to conventional whole breast radiotherapy for selected patients. Recently, CyberKnife has emerged as a possible alternative to conventional techniques for accelerated partial breast irradiation. In this retrospective study, we present a dosimetric comparison between 3-dimensional conformal radiotherapy plans and CyberKnife plans using circular (Iris) and multi-leaf collimators. Nine patients who had undergone breast-conserving surgery followed by whole breast radiation were included in this retrospective study. The CyberKnife planning target volume (PTV) was defined as the lumpectomy cavity + 10 mm + 2 mm with prescription dose of 30 Gy in 5 fractions. Two sets of 3-dimensional conformal radiotherapy plans were created, one used the same definitions as described for CyberKnife and the second used the RTOG-0413 definition of the PTV: lumpectomy cavity + 15 mm + 10 mm with prescription dose of 38.5 Gy in 10 fractions. Using both PTV definitions allowed us to compare the dose delivery capabilities of each technology and to evaluate the advantage of CyberKnife tracking. For the dosimetric comparison using the same PTV margins, CyberKnife and 3-dimensional plans resulted in similar tumor coverage and dose to critical structures, with the exception of the lung V5%, which was significantly smaller for 3-dimensional conformal radiotherapy, 6.2% when compared to 39.4% for CyberKnife-Iris and 17.9% for CyberKnife-multi-leaf collimator. When the inability of 3-dimensional conformal radiotherapy to track motion is considered, the result increased to 25.6%. Both CyberKnife-Iris and CyberKnife-multi-leaf collimator plans demonstrated significantly lower average ipsilateral breast V50% (25.5% and 24.2%, respectively) than 3-dimensional conformal radiotherapy (56.2%). The CyberKnife plans were more conformal but less homogeneous than the 3-dimensional conformal radiotherapy plans. Approximately 50% shorter treatment times and 50% lower number of delivered monitor units (MU) were achievable with CyberKnife-multi-leaf collimator than with CyberKnife-Iris. The CyberKnife-multi-leaf collimator treatment times were comparable to 3-dimensional conformal radiotherapy, however, the number of MU delivered was approximately 2.5 times larger. The suitability of 10 + 2 mm margins warrants further investigation.