Near-field optical and atomic force constraints for super-resolution 3D deconvolution in far-field optical microscopy

We demonstrate that near-field optical and atomic force microscopy data can be used for super-resolution three-dimensional (3-D) image restoration in optical sectioning fluorescence microscopy. A crucial feature in this approach is the full integration of such data sets with digital far-field images. The scanned probe data is used to provide modalities for boundary constraints which define surface optical information and spatial domains of optical alterations in a sample with a spatial precision that has been unachievable in the past. A restoration algorithm that can use such a complex of data for 3-D image deconvolution is presented. It uses a Tikhonov-Miller regularization scheme and allows for the imposition of different types of constraints to obtain super-resolution deconvolved images. Performance was tested by using simulated 3-D imaging. An example is given of the restoration of a 3-D wide field optical image of a biological specimen in the presence of atomic force constraints.