US 12,265,033 B2 · 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 (S1) 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 (S2) with one light distribution. For each of the light distributions, photons emitted by the molecules are detected (S4) for different positionings of the light distribution. The light distributions are positioned (S3) 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 (S5).

Claims25

Dependent claims

A method for determining positions of two or more spaced-apart molecules in one or more spatial directions in a sample using a localization microscope, wherein light distributions arising due to interference of coherent light are used for determining the positions of the molecules, comprising:

generating a plurality of light distributions using a first light modulator having a plurality of switchable pixels disposed in an image plane of the localization microscope, each light distribution having at least one local intensity minimum and regions with an intensity increase adjacent thereto;
illuminating each of the two or more molecules with one light distribution each;
for each of the light distributions, detecting photons emitted by the molecules for different positionings of the light distribution, the light distributions being positioned independently of each other;
and
deriving the positions of the molecules based on the photons detected for the different positionings of the light distributions.

2 more elements

An apparatus for determining positions of two or more spaced-apart molecules in one or more spatial directions in a sample using a localization microscope, wherein light distributions arising due to interference of coherent light are used for determining the positions of the molecules, comprising:

an operating unit adapted to drive a first light modulator having a plurality of switchable pixels, which is disposed in an image plane of the localization microscope, to generate a sequence of light distributions, each light distribution having local intensity minima and regions with an intensity increase adjacent thereto, wherein subsequent light distributions are locally modified in accordance with estimated positions of the molecules;
and
an evaluation unit adapted to derive the positions of the molecules on the basis of photons emitted by the molecules for the different light distributions.

Dependencies are inferred from the claim text.

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

Life of this patent

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

Life of this patent

Abstract

Claims25