Abstract: A light-microscopy method for locating point objects in a sample arranged in an object space includes imaging the sample onto a detector by an imaging optical unit having a depth of field of predetermined axial extent along an optical axis in the object space, onto which the detector is imaged. The point objects in the sample are located within the depth of field. The first sample image generated by the imaging of the sample onto the detector is evaluated. For locating a respective first point object in a direction of the optical axis, a parameter of a first light spot of one or more light spots of the first sample image representing the first point object is determined, and a rough axial z position related to the first point object is assigned to the parameter based on predetermined association information.

Abstract: A widefield microscope illumination system and a method for illumination, the system having a microscope objective with an optical objective axis, an illumination light source sending widefield illumination light along illumination beam paths having corresponding illumination axes along which the illumination light penetrates into the microscope objective through illumination light entry sites located within a predetermined illumination light entry area, a spatially resolving light detector detecting detected light sent from an illuminated sample through the microscope objective along a detected light beam path, and an automatic illumination light beam path manipulation device, controlled by a control system, which is arranged in front of the microscope objective in relation to the direction of the illumination light beam path, and by which illumination light beam path manipulation device the illumination axes are automatically movable at time intervals to a plurality of illumination light entry sites.

Abstract: A microscopic device for three-dimensional localization of point-like objects, encompassing a detection optical system that images point-like objects, each in the form of a three-dimensional focus light distribution, into an image space; a color separation apparatus that divides the light into at least two separate light bundles of different wavelength regions; at least two image space detector units , one receiving one light bundle and the other receiving the other light bundle, each detector unit comprising a light-spot-sensing detection surface ; an evaluation unit that ascertains a lateral X-Y position and an axial Z position relative to the sharpness plane in a direction perpendicular to the sharpness plane; at least one Z-position correction value for at least one of the wavelength regions being stored in the evaluation unit, which value indicates a detection optical system longitudinal chromatic aberration in that wavelength region; and the evaluation unit correcting the Z position.

Abstract: A sample retainer for a microscope is described, comprising a sample stage (32), a holder (34) arranged on the sample stage (32), a sample carrier (36), couplable to the holder (34), to which a sample is attachable, and an adjusting apparatus (44), engaging on the holder (34), with which with the sample carrier (36), together with the holder (34) to which the sample carrier (36) is coupled, is displaceable on the sample stage (32), relative to the objective (46), into a target position. A decoupling apparatus that decouples the sample carrier (36), arranged in the target position, from the holder (34) upon imaging of the sample through the objective (46) is provided.

Abstract: A method and an apparatus for imaging a sample structure (34) with a light microscope are described. The sample structure (34) is prepared with markers and an active marker subset is generated in that a part of the markers is brought into a state in which they can be imaged. When a predetermined activation state is present, the sample structure (34) is imaged onto an arrangement (40) of sensor elements which each generate an image signal, these image signals resulting in a single frame of the sample structure (34). The presence of a predetermined activation state is checked in that at least two test single frames are generated at an interval, and the respective change of the image signal between the two test single frames is detected. The presence of the predetermined activation state is determined when the detected changes of the image signals in their entirety exceed a quantity.

Abstract: A widefield microscope illumination system and a method for illumination, the system having a microscope objective with an optical objective axis, an illumination light source sending widefield illumination light along illumination beam paths having corresponding illumination axes along which the illumination light penetrates into the microscope objective through illumination light entry sites located within a predetermined illumination light entry area, a spatially resolving light detector detecting detected light sent from an illuminated sample through the microscope objective along a detected light beam path, and an automatic illumination light beam path manipulation device, controlled by a control system, which is arranged in front of the microscope objective in relation to the direction of the illumination light beam path, and by which illumination light beam path manipulation device the illumination axes are automatically movable at time intervals to a plurality of illumination light entry sites.

Abstract: A microscope stage (14) comprising a platform (16), a specimen holder (18) resting on the platform (16) and a positioning device (20) for moving the specimen holder (18) in a plane of displacement parallel to the platform (16) is described. The microscope stage (14) includes a positioning device (20) having two displacing devices (34, 36) which are mechanically decoupled from each other and of which a first displacing device (34) is designed to move the specimen holder (18) along a first axis in the plane of displacement, and a second displacing device (36) is designed to move the specimen holder (18) along a second axis in the plane of displacement, which second axis runs transversely to the first axis.

Abstract: A device and a method for acquiring a microscopic image of a sample structure are described. An optic for imaging the sample structure and a reference structure is provided, as well as a drift sensing unit for sensing a drift of the sample structure relative to the optic on the basis of the imaged reference structure. The optic comprises a first sharpness plane for imaging the sample structure and at the same time a second sharpness plane, modifiable in location relative to the first sharpness plane, for imaging the reference structure.

Abstract: A sample retainer for a microscope is described, comprising a sample stage (32), a holder (34) arranged on the sample stage (32), a sample carrier (36), couplable to the holder (34), to which a sample is attachable, and an adjusting apparatus (44), engaging on the holder (34), with which with the sample carrier (36), together with the holder (34) to which the sample carrier (36) is coupled, is displaceable on the sample stage (32), relative to the objective (46), into a target position. A decoupling apparatus that decouples the sample carrier (36), arranged in the target position, from the holder (34) upon imaging of the sample through the objective (46) is provided.

Abstract: A sequence of individual images is acquired by imaging the sample through imaging optics onto an image sensor. For the acquisition of each individual image, the sample is provided with a marker pattern, in which individual markers can be imaged in the form of spatially separable light distributions through the imaging onto the image sensor. The centroid positions of the light distributions are determined and superimposed to form a complete image of the sample. According to the present invention, an image-drift-inducing temperature value (?T1, ?T2, . . . , ?Tn) is measured during the acquisition of the sequence of individual images. A temperature-dependent drift value (?X1, ?X2, . . . , ?Xn; ?Y1, ?Y2, . . . , ?Yn) is correlated to the image-drift-inducing temperature value (?T1, ?T2, . . . , ?Tn) based on predetermined correlation data. The determined centroid positions are corrected based on the drift value (?X1, ?X2, . . . , ?Xn; ?Y1, ?Y2, . . . , ?Yn).

Abstract: A method for setting an evaluation parameter for a fluorescence microscope includes exciting dye particles in a sample to fluoresce and detecting fluorescent light from the particles. A graphical representation of a distribution of the fluorescent light is determined and a signal is generated for use in displaying the graphical representation on a display unit. Each subregion of the graphical representation is associated with a comparison value that is representative of a light quantity in the subregion. A predefined threshold is used as an evaluation parameter and compared to the comparison values. The subregions having a comparison value that is greater than the threshold value are marked on the display unit with predefined markings. The threshold value is changed and the comparison values are compared to the changed threshold value. The marked regions are defined as events and a complete image of the sample is obtained based on the events.

Abstract: A microscope stage (14) comprising a platform (16), a specimen holder (18) resting on the platform (16) and a positioning device (20) for moving the specimen holder (18) in a plane of displacement parallel to the platform (16) is described. The microscope stage (14) includes a positioning device (20) having two displacing devices (34, 36) which are mechanically decoupled from each other and of which a first displacing device (34) is designed to move the specimen holder (18) along a first axis in the plane of displacement, and a second displacing device (36) is designed to move the specimen holder (18) along a second axis in the plane of displacement, which second axis runs transversely to the first axis.

Abstract: In high spatial resolution imaging, a structure in a specimen is marked with a substance which, in a first electronic state, is excited by light of one wavelength to emit fluorescent light, which is also converted from its first into a second electronic state by that light, and which returns from its second into its first electronic state. The specimen is imaged onto a sensor at a spatial resolution not resolving an average spacing between neighboring molecules of the substance, and exposed to the light at such an intensity that the molecules in the first state are alternately excited to emit fluorescent light and converted into their second state, and that at least 10% of the molecules presently in their first state lie at a distance from their closest neighboring molecules in their first state which is greater than the spatial resolution of the imaging onto the sensor.

Abstract: A method and an apparatus for imaging a sample structure (34) with a light microscope are described. The sample structure (34) is prepared with markers and an active marker subset is generated in that a part of the markers is brought into a state in which they can be imaged. When a predetermined activation state is present, the sample structure (34) is imaged onto an arrangement (40) of sensor elements which each generate an image signal, these image signals resulting in a single frame of the sample structure (34). The presence of a predetermined activation state is checked in that at least two test single frames are generated at an interval, and the respective change of the image signal between the two test single frames is detected. The presence of the predetermined activation state is determined when the detected changes of the image signals in their entirety exceed a quantity.

Abstract: For imaging of a structure, the structure is marked with a substance which can be converted by a switching signal from a first into a second state, and which provides an optical measurement signal in one of its states, only. The switching signal is applied such that at least 10% of the molecules of the substance being in the measurement signal providing state are at a distance from their closest neighbors, which is greater than the spatial resolution limit of imaging the specimen onto a sensor array, which in turn is greater than an average distance between the molecules of the substance. From an intensity distribution of the measurement signal recorded with the sensor array, the position is only determined for those molecules of the substance which are at a distance from their closest neighboring molecules in the measurement signal providing state, which is greater than the spatial resolution limit.

Abstract: For imaging of a structure, the structure is marked with a substance which can be converted by a switching signal from a first into a second state, and which provides an optical measurement signal in one of its states, only. The switching signal is applied such that at least 10% of the molecules of the substance being in the measurement signal providing state are at a distance from their closest neighbors, which is greater than the spatial resolution limit of imaging the specimen onto a sensor array, which in turn is greater than an average distance between the molecules of the substance. From an intensity distribution of the measurement signal recorded with the sensor array, the position is only determined for those molecules of the substance which are at a distance from their closest neighboring molecules in the measurement signal providing state, which is greater than the spatial resolution limit.

Abstract: In high spatial resolution imaging, a structure in a specimen is marked with a substance which, in a first electronic state, is excited by light of one wavelength to emit fluorescent light, which is also converted from its first into a second electronic state by that light, and which returns from its second into its first electronic state. The specimen is imaged onto a sensor at a spatial resolution not resolving an average spacing between neighboring molecules of the substance, and exposed to the light at such an intensity that the molecules in the first state are alternately excited to emit fluorescent light and converted into their second state, and that at least 10% of the molecules presently in their first state lie at a distance from their closest neighboring molecules in their first state which is greater than the spatial resolution of the imaging onto the sensor.

Abstract: For the high spatial resolution imaging of a structure of interest in a specimen, a substance is selected from a group of substances which have a fluorescent first state and a nonfluorescent second state; which can be converted fractionally from their first state into their second state by light which excites them into fluorescence, and which return from their second state into their first state; the specimen's structure of interest is imaged onto a sensor array, a spatial resolution limit of the imaging being greater (i.e.