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...It is capable of minimizing the systematic error that occurs with astigmatism-based 3D techniques owing to the aberrations induced by the refractive index mismatch. In contrast to existing techniques, the method determines the aberration directly from the acquired 2D image stream by exploiting the inherent particle movement and the redundancy introduced by the astigmatism. It does not require additional experimental effort for the user and imaging can directly be started once interesting regions in the sample have been identified. MDA is written in MATLAB and comes with a graphical user interface. The particle fitting routine is provided by an external library written in C. ...