Abstract: The invention relates to a method for localizing individual emitters in a sample, comprising a pre-localization comprising an illumination of the sample with illumination light, wherein the illumination light induces or modulates light emissions of an individual and stationary emitter in the sample, detecting the light emissions of the emitter and estimating the position of the emitter in the sample from the detected light emissions and a subsequent main localization comprising illuminating the sample with an intensity distribution of the illumination light at illumination positions, the intensity distribution comprising a local minimum, detecting the light emissions of the emitter for the illumination positions, and determining the position of the emitter in the sample from the light emissions detected for the illumination positions, wherein the illumination positions are arranged in a first iteration about the estimated position and wherein the illumination positions in at least one second iteration are arr

Abstract: Methods and a device for imaging a sample stained with multiple dyes are disclosed. The methods enable nanoscopic imaging with up to molecular resolution to be placed in the spatial context of microscopic images, or nanoscopic tracking of individual molecules to be performed in the spatial context of a microscopic image. The methods combine different light-optical microscopy techniques in a particularly efficient manner. Molecular resolution can be achieved by a localization microscopic method, in particular by localization according to a MINFLUX or STED-MINFLUX method. These methods are characterized by the fact that, on the one hand, they are particularly gentle on the sample during the steps that precede imaging with molecular resolution, and on the other hand, they make optimal use of the fluorescence photons during localization or enable optimal use.

Abstract: An apparatus is provided for monitoring a focal state of a microscope having an object plane and a main imaging area. The apparatus has an auxiliary light source providing an auxiliary light beam and coupling the auxiliary light beam into the microscope in such a way that the coupled auxiliary light beam runs within a plane which is spanned outside of the main imaging area by a straight line running in the object plane and a normal to the object plane, and that the coupled auxiliary light beam is inclined at an angle to a normal to the object plane. A part of the coupled auxiliary light beam reflected by a reference boundary surface in the microscope impinges on a registration device in an area of incidence. The registration device registers position changes of the area of incidence on the registration device.

Abstract: The present disclosure relates to localisation microscopic investigations of samples stained with multiple dyes. According to the present disclosure, it is either provided that a singulated excitable fluorophore of a first species is excited with excitation light of two different wavelengths and that a localisation of the fluorophore is obtained for each of the two wavelengths, or that first test excitation is performed in order to then select a wavelength for excitation light with which a singulated fluorophore is localised. In the first case, the difference in the localisations of the one or preferably multiple individual fluorophores obtained in this way is determined and used to obtain localisations of fluorophores of a different species and those of the first species in a common spatial reference system. The second case is advantageously applicable for tracking structures. Also in the second case, the advantages of the first case can be additionally realised.

Abstract: The invention relates to a method for localizing an emitter (F) in a sample (S) comprising illuminating the sample (S) with a stationary donut-shaped excitation beam (E), acquiring fluorescence photons; and estimating a position of the emitter (F) in the sample (S) from the acquired fluorescence photons. The invention further relates to an apparatus (1) for localizing an emitter (F) in a sample (S) comprising illumination means (10), acquisition means (20) and processing means (30) and a computer program comprising instruction to cause the apparatus (1) to execute the method for localizing an emitter (F).

Abstract: The invention relates to a method for determining positions of mutually spaced molecules (M) in a sample (20), having the steps of generating (101) a plurality of light distributions, each light distribution having a local intensity minimum (110, 310) and adjacent regions (120, 320) of increasing intensity, comprising an excitation light distribution (100) and a deactivation light distribution (300); illuminating (102) the sample (20) with the excitation light distribution (100) and the deactivation light distribution (300); detecting (103) photons emitted by the molecule (M) for different positions of the excitation light distribution (100); and deriving (104) the position of the molecule (M) on the basis of the photons detected for the different positions of the excitation light distribution (100), wherein the local minimum (110) of the excitation light distribution (100) is arranged at a plurality of scanning positions (201) one after the other within a scanning region (200), and the light intensity of the

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.

Abstract: An apparatus is provided for monitoring a focal state of a microscope having an object plane and a main imaging area The apparatus has an auxiliary light source providing an auxiliary light beam and coupling the auxiliary light beam into the microscope in such a way that the coupled auxiliary light beam runs within a plane which is spanned outside of the main imaging area by a straight line running in the object plane and a normal to the object plane, and that the coupled auxiliary light beam is inclined at an angle to a normal to the object plane. A part of the coupled auxiliary light beam reflected by a reference boundary surface in the microscope impinges on a registration device in an area of incidence. The registration device registers position changes of the area of incidence on the registration device.

Abstract: An optical filter array includes a substrate permeable to an electromagnetic radiation to be detected, a first DBR mirror arranged on the substrate, a second DBR minor arranged above the first DBR mirror, and a plurality of cavity sections. The cavity sections have different respective optical lengths, and are arranged so as to be spatially separated from each other between the first DBR mirror and the second DBR mirror. Each of the first DBR mirror, the second DBR mirror, and the plurality of cavity sections with different optical lengths form filter elements of a filter. The filter reflects in a stopband determined by the first DBR mirror and the second DBR mirror. Each filter element has at least one narrow transmission band determined by the optical length of its respective cavity section located inside the stopband. A different thicknesses of the cavity sections is provided via a nanoimprint process.

Abstract: An optical filter array includes a substrate permeable to an electromagnetic radiation to be detected, a first DBR mirror arranged on the substrate, a second DBR minor arranged above the first DBR mirror, and a plurality of cavity sections. The cavity sections have different respective optical lengths, and are arranged so as to be spatially separated from each other between the first DBR minor and the second DBR mirror. Each of the first DBR minor, the second DBR mirror, and the plurality of cavity sections with different optical lengths form filter elements of a filter. The filter reflects in a stopband determined by the first DBR mirror and the second DBR mirror. Each filter element has at least one narrow transmission band determined by the optical length of its respective cavity section located inside the stopband. A different thicknesses of the cavity sections is provided via a nanoimprint process.

Abstract: The invention relates to an optical filter and a method for its production, and to a device for the examination of the spectral and spatial distribution of an electromagnetic radiation irradiated from an object. The invention is based on the task of providing an optical filter of the above described type that is inexpensive to produce, which can be used to detect a plurality of wavelengths, in which, however, tuning of the DBR mirrors by means of displacement is not necessary. Furthermore, a method for the production of such a filter is provided.

Abstract: The invention relates to an optical filter and a method for its production, and to a device for the examination of the spectral and spatial distribution of an electromagnetic radiation irradiated from an object. The invention is based on the task of providing an optical filter of the above described type that is inexpensive to produce, which can be used to detect a plurality of wavelengths, in which, however, tuning of the DBR mirrors by means of displacement is not necessary. Furthermore, a method for the production of such a filter is provided.