Method, computer program and apparatus for determining positions of molecules in a sample

Abstract

The present disclosure relates to method, computer programs with instructions, and apparatus for determining positions of two or more spaced-apart molecules in one or more spatial directions in a sample by means of a localization microscope. The present disclosure also relates to localization microscopes using such an apparatus. Light distributions arising due to interference of coherent light are used for determining the positions of the molecules. In the method, a plurality of light distributions are generated (S) using a first light modulator having a plurality of switchable pixels. The first light modulator is arranged in an image plane of the localization microscope. Each light distribution has a local intensity minimum and regions with an intensity increase adjacent thereto. Each of the two or more molecules is illuminated (S) with one light distribution. For each of the light distributions, photons emitted by the molecules are detected (S) for different positionings of the light distribution. The light distributions are positioned (S) independently of each other. Based on the photons detected for the different positionings of the light distributions, the positions of the molecules are finally derived (S).