Compact fluorescence depletion microscope system using an integrated optical element

The performance of a fluorescence depletion microscope system depends critically on the precise alignment of both the pump and the erase beams with the axis of the focusing objective lens. In this article we propose a technique with which the need for separate alignment of the pump and erase beams is eliminated. Besides being insensitive to alignment errors, our technique is much more compact than present systems, and can serve as a basis for a commercial fluorescence depletion microscope.