A digital holographic technique is implemented in a microscope for three-dimensional imaging reconstruction. The setup is a Mach–Zehnder interferometer that uses an incoherent light source to remove the coherent noise that is inherent in the laser sources. A phase-stepping technique determines the optical phase in the image plane of the microscope. Out-of-focus planes are refocused by digital holographic computations, thus considerably enlarging the depth of investigation without the need to change the optical focus mechanically. The technique can be implemented in transmission for various magnification ratios and can cover a wide range of applications. Performances and limitations of the microscope are theoretically evaluated. Experimental results for a test target are given, and examples of two applications in particle localization and investigation of biological sample are provided.
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