A method for the spatially high-resolution determining the location of a separated, with excitation light to the emission of luminescence light stimulable molecule in a sample

摘要

In the case of a method for the spatially high-resolution determining the location (x₀) a single molecule in one or more directions in space in a sample, wherein said molecule with excitation light to the emission of luminescence light (12) is excitable, the anregungslichtmit an intensity distribution is directed onto the sample, which in each of the spatial directions at least one intensity increase in area (22) with a known a strictly monotonic course of the intensity of the excitation light over a distance to a reference point has the intensity distribution. An initial area (23) in the sample, in which said molecule is arranged, is determined. (i) in each of the directions in space, is at least one position (x_N1) the application point of the intensity distribution is set such that the at least one intensity increase in area (22) in the respective spatial direction by means of the initial area (23); the top point of the intensity distribution is at the positions (x_N1) in the sample and arranged at each position (x_N1) the application point of the intensity distribution in the sample, the luminescence light emitted by the molecule (12). (ii) from the intensity values (i₁; I_Sw) the luminescence light (12), which leads to each of the spatial directions two intensity values (i₁; I_Sw) , one of which the intensity (i₁) of the to the at least one position (x_N1) the application point of the intensity distribution registered luminescence light (12) is a further area (24) in the sample is determined, in which said molecule is arranged and the smaller than the initial area (23) is. The steps of (i) and (ii) are at least once using of the further location area (24) as a new initial area (23) is repeated.n="45"