Ground-state-depletion fluorscence microscopy: A concept for breaking the diffraction resolution limit

Abstract

We introduce and study a novel concept in farfield fluorescence microscopy fundamentally overcoming the classical diffraction resolution limit. This is accomlished by reducing the spatial extent of the effective focus of a scanning fluorescence microscope. The reduction is achieved by depleting the ground-state energy of the molecules located in the outer region of the focus. Our theoretical study shows that ground-state-depletion fluorescence microscopy has the potential of increasing the resolution of far-field fluorescence microscopy by an order of magnitude which is equivalent to a lateral resolution of 15 NM. © 1995 Springer-Verlag.