Abberior Instruments GmbH

The present invention is a method for spatially highly accurate location determination of individual dye molecules of a fluorescent dye by scanning with an intensity distribution of a scanning light having a local minimum. The invention is characterized by the fact that the scanning is not performed uniformly for all dye molecules, but is individually adapted to the dye molecule to be scanned and, if necessary, to its environment in the sample, in order to achieve the most accurate location determination possible with the smallest possible number of fluorescence photons.

The invention relates to a method for determining a reference time point (to) by means of a light microscope ( 1 ), wherein a sample ( 2 ) is illuminated by first light pulses (P 1 ) in order to excite light-emitting entities in the sample ( 2 ), wherein a light signal generated by means of the first light pulses (P 1 ) is detected, and wherein the reference time point (to) is determined based on the detected light signal, and wherein the light signal is detected in at least two measurements each in a first detection time window (G 1 ), wherein a starting time point of the first detection time window (G 1 ) is adjusted for each of the measurements. The invention further relates to a light microscope ( 1 ) and a computer program for carrying out the method.

The present disclosure relates to MINFLUX nanoscopy. The present disclosure improves three-dimensional localization of isolated emitters, particularly of isolated fluorescent emitters. The utilization of the emitted photons of isolated excitable emitters, in particular of isolated excitable fluorescent emitters, for a three-dimensional localization is improved by separating the localization according to the MINFLUX principle using a 3D excitation donut into a sequence of separate steps, wherein in one step either an axial location or a lateral location of the fluorophore is determined. Thereby, the more precise knowledge of the axial position increases the quality of a following lateral localization and vice versa. The three-dimensional localization is further improved by a method for axial localization by scanning an axial scanning area with a 3D excitation donut and a preferably following evaluation of the measured intensities or photon numbers by forming a vector sum.

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).

A high-resolution microscope comprises a carrier support, a plurality of beam guiding and beam forming optical elements mounted to the carrier support in a defined spatial arrangement, and a housing covering the optical elements mounted to the carrier support. The housing comprises a housing panel. The housing panel has two parallel facesheets and a core layer. The core layer is bonded to the two facesheets and includes at least one of a plurality of gas filled cavities and a heavy layer such that the housing panel prevents airborne sound and air flows that occur in the environment of the microscope from exciting the optical elements to vibrations.

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 deactivation light in a catching region ( 210 ), which is paired with the scanning region ( 200 ) and in which the position of the molecule (M) can be unambiguously derived from the scanning positions ( 201 ) and the paired detected photons, corresponds maximally to three times the saturation intensity of the deactivation light. The invention further relates to a device for carrying out said method.

The invention relates to a sensor circuit for a device performing a safety function, comprising at least two sensors, at least two comparison circuits, each of the comparison circuits being assigned to one of the sensors, and a linking unit for combining logic states (L=0, L=1) of comparison circuit outputs of the comparison circuits to form a circuit output signal, the sensor circuit being configured to scale the comparison circuit output value of at least a first one of the comparison circuits and to feed it back to a measurement input or a reference input of at least a second one of the comparison circuits so that, when the comparison circuit output of the first comparison circuit transitions between the logic states (L=0, L=1), the difference between the measurement signal and the reference signal of the second comparison circuit is reduced or the sign of the difference between the measurement signal and the reference signal of the second comparison circuit is reversed. The invention further relates to a device comprising the sensor circuit and a method for processing measurement values from sensors.

An auxiliary apparatus for testing the confocality of a scanning and descanning microscope component group has a connector configured for connecting the auxiliary apparatus in a defined relative position to the scanning and descanning microscope component group, and an optical axis running at a fixed orientation with respect to the connector. Further, the auxiliary apparatus has an auxiliary detector with a plurality of auxiliary detection apertures in a plurality of auxiliary detection aperture positions that are arranged at distances in direction of the optical axis and laterally with respect to the optical axis; and an auxiliary light source providing auxiliary light through a plurality of auxiliary emission apertures in a plurality of auxiliary emission aperture positions arranged at distances in direction of the optical axis and laterally with respect to the optical axis.

The invention relates to a detection device ( 2 ) for a laser scanning microscope, the detection device ( 2 ) having a light inlet ( 4 ), at least one filter module ( 14 ) and at least one spatially resolving detector ( 22 ) and being configured to guide light from the light inlet ( 4 ) to the filter module ( 14 ) and from there to the spatially resolving detector ( 22 ), at least one filter module ( 14 ) being designed as a continuous filter module with two continuously tunable filter elements ( 16 ), and at least one compensator element ( 26 ) being arranged optically downstream of the continuous filter module ( 14 ), by means of which a focal position of light on the spatially resolving detector ( 22 ) can be adjusted.

The present disclosure relates to methods for generating high-resolution images of a structure in a sample or for localizing individual molecules of a fluorescent dye in a sample, and the use of a fluorescent dye in such a method. The methods according to the present disclosure are characterized in that the fluorescent dye is first formed from a protected, non-fluorescent form of the dye in a photoactivation reaction comprising at least two reaction steps prior to scanning with excitation and fluorescence inhibition light, and that the protected, non-fluorescent form of the dye is inert to the excitation and fluorescence inhibition light.