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Focus on Microscopy 2014 is the continuation of a yearly conference series presenting the latest innovations in optical microscopy and their application in biology, medicine and the material sciences. Key subjects for the conference series are the theory and practice of 3D optical imaging, related 3D image processing, and reporting especially on developments in resolution and imaging modalities. The conference series covers also the rapidly advancing fluorescence labeling techniques for confocal and multi-photon 3D imaging of -live- biological specimens.