The study of a new family of phase masks for three-dimensional fluorescence nanoscopy

Abstract

The question of the axial coordinate determination accuracy is quite sophisticated in far field 3D fluorescence nanoscopy. The accuracy of a point emitter axial coordinate reconstruction depends among other things on the conversion efficiency of the phase mask and the microscope objective instrumental function. It was found that the instrumental functions of different microscope objectives differ significantly from each other, most have a strongly non-uniform spatial distribution of the radiation intensity in a parallel beam created by an objective focused on a point emitter. It was shown that taking into account the actual microscope objective instrumental function when calculating the phase masks allows to increase significantly the axial coordinate reconstruction accuracy.