Abstract: For high spatial resolution imaging a structure marked with luminescence markers, light that has an effect on the emission of luminescence light by the luminescence markers is directed onto a sample with an intensity distribution having a central zero point. Scan areas of the sample are scanned with the zero point. Luminescence light emitted out of a local area including the zero point is registered and assigned to the respective location of the zero point in the sample. Several copies of an object of interest are arranged in the scan areas and subjected to varying surrounding conditions. The individual scan areas are scanned with the respective zero point at least two times at two different stages of reactions to the varying surrounding conditions. Dimensions of the scan areas are limited such that they are not larger than 75% of a distance of intensity maxima delimiting the zero point.

Abstract: For high spatial resolution imaging a structure marked with luminescence markers, light that has an effect on the emission of luminescence light by the luminescence markers is directed onto a sample with an intensity distribution having a central zero point. Scan areas of the sample are scanned with the zero point. Luminescence light emitted out of a local area including the zero point is registered and assigned to the respective location of the zero point in the sample. Several copies of an object of interest are arranged in the scan areas and subjected to varying surrounding conditions. The individual scan areas are scanned with the respective zero point at least two times at two different stages of reactions to the varying surrounding conditions. Dimensions of the scan areas are limited such that they are not larger than 75% of a distance of intensity maxima delimiting the zero point.

Abstract: For high spatial resolution imaging a structure in a sample, the structure being marked with luminescence markers, light that has an effect on the emission of luminescence light by the luminescence markers is directed onto the sample with an intensity distribution having a zero point and intensity maxima neighboring the zero point in at least one direction. A scan area which is a part of the sample is scanned with the zero point. Luminescence light emitted out of a local area including the zero point is registered and assigned to the respective location of the zero point in the sample. Dimensions of the scan area, in at least one direction in which the intensity maxima are neighboring the zero point, are limited such that they are not larger than 75% of a distance of the intensity maxima in the at least one direction.

Abstract: For high spatial resolution imaging a structure in a sample, the structure being marked with luminescence markers, light that has an effect on the emission of luminescence light by the luminescence markers is directed onto the sample with an intensity distribution having a zero point and intensity maxima neighboring the zero point in at least one direction. A scan area which is a part of the sample is scanned with the zero point. Luminescence light emitted out of a local area including the zero point is registered and assigned to the respective location of the zero point in the sample. Dimensions of the scan area, in at least one direction in which the intensity maxima are neighboring the zero point, are limited such that they are not larger than 75% of a distance of the intensity maxima in the at least one direction.

Abstract: In a STED fluorescence light microscope pulses of excitation light (3) are applied to a sample, which excite fluorescent entities contained in the sample for fluorescence, and which are focused on at least one focal area. Further, de-excitation light (12) is applied to the sample, which de-excites the excited fluorescent entities and which comprises an intensity zero point in the at least one focal area, as a continuous wave. Fluorescence light emitted by the excited fluorescent entities in the sample is registered after each pulse of the excitation light (3) and overlapping with applying the de-excitation light (13) with high temporal resolution between consecutive pulses of the excitation light (3).

Abstract: In an optical assembly having a light source which provides two optically different light components with essentially planar wavefronts on an optical axis, wherein the light components differ at least in their wavelength; in the case of an objective lens which projects the two optically different light components into a projection space; and in the case of an optical component which is arranged on the optical axis and has an plane through which the wavefronts of the two light components pass and in which at least two different areas of the optical component with different dispersion behaviors n(?) abut against one another in the lateral direction with respect to the optical axis; the optical component causes phase shifts of the wavefronts of the two light components, wherein the phase shift of the wavefronts of the one light component differs by at least one quarter of the wavelength of that light component between the two different areas, and wherein the phase shift of the wavefronts of the other light compo

Abstract: The invention relates to novel and improved photostable rhodamine dyes of the general structural formulae I or II and their uses as fluorescent markers, e.g. for immunostainings and spectroscopic and microscopic applications, in particular in conventional and stimulated emission depletion (STED) microscopy and fluorescence correlation spectroscopy.

Abstract: In a STED fluorescence light microscope pulses of excitation light (3) are applied to a sample, which excite fluorescent entities contained in the sample for fluorescence, and which are focused on at least one focal area. Further, de-excitation light (12) is applied to the sample, which de-excites the excited fluorescent entities and which comprises an intensity zero point in the at least one focal area, as a continuous wave. Fluorescence light emitted by the excited fluorescent entities in the sample is registered after each pulse of the excitation light (3) and overlapping with applying the de-excitation light (13) with high temporal resolution between consecutive pulses of the excitation light (3).

Abstract: The invention relates to novel and improved photostable rhodamine dyes of the general structural formulae I or II and their uses as fluorescent markers, e.g. for immunostainings and spectroscopic and microscopic applications, in particular in conventional and stimulated emission depletion (STED) microscopy and fluorescence correlation spectroscopy. The partially deuterated analogues are useful as molecular mass distribution tags in mass spectroscopic applications.

Abstract: A method of producing spatial fine structures comprises the steps of: selecting a luminophore from the group of luminophores displaying two different states, one of the two states being an active state in which luminescence light is obtainable from the luminophore, the other of the two states being an inactive state in which no luminescence light is obtainable from the luminophore, and the luminophore being reversibly, but essentially completely, transferable out the one state into the other state by means of an optical signal; adding the luminophore to a material; forming a spatial fine structure of the material; and fluorescence-microscopically examining whether the desired fine structure is present.

Abstract: In an optical assembly having a light source which provides two optically different light components with essentially planar wavefronts on an optical axis, wherein the light components differ at least in their wavelength; in the case of an objective lens which projects the two optically different light components into a projection space; and in the case of an optical component which is arranged on the optical axis and has an plane through which the wavefronts of the two light components pass and in which at least two different areas of the optical component with different dispersion behaviors n(?) abut against one another in the lateral direction with respect to the optical axis; the optical component causes phase shifts of the wavefronts of the two light components, wherein the phase shift of the wavefronts of the one light component differs by at least one quarter of the wavelength of that light component between the two different areas, and wherein the phase shift of the wavefronts of the other light compo

Abstract: A method of microscopically examining a spatial fine structure comprises the steps of selecting a luminophore from the group of luminophores which have two physical states, the two states differing from each other with regard to the luminescence properties displayed by the luminophore, and which are reversibly, but essentially completely transferable out of one into the other state of their two states by means of an optical signal; overlaying a surface of the spatial fine structure with the luminophore; and determining the profile of the surface overlaid with the luminophore. The step of determining the profile of the surface comprises the sub-steps of transferring the luminophore by means of the optical signal out of the one into the other of its two states outside a presently observed measurement point, measuring luminescence light emitted by the luminophore, and repeating the sub-steps of transferring and measuring for further measurement points distributed over the surface.

Abstract: A method of producing spatial fine structures comprises the steps of: selecting a luminophore from the group of luminophores displaying two different states, one of the two states being an active state in which luminescence light is obtainable from the luminophore, the other of the two states being an inactive state in which no luminescence light is obtainable from the luminophore, and the luminophore being reversibly, but essentially completely, transferable out the one state into the other state by means of an optical signal; adding the luminophore to a material; forming a spatial fine structure of the material; and fluorescence-microscopically examining whether the desired fine structure is present.

Abstract: A method of microscopically examining a spatial fine structure comprises the steps of selecting a luminophore from the group of luminophores which have two physical states, the two states differing from each other with regard to the luminescence properties displayed by the luminophore, and which are reversibly, but essentially completely transferable out of one into the other state of their two states by means of an optical signal; overlaying a surface of the spatial fine structure with the luminophore; and determining the profile of the surface overlaid with the luminophore. The step of determining the profile of the surface comprises the sub-steps of transferring the luminophore by means of the optical signal out of the one into the other of its two states outside a presently observed measurement point, measuring luminescence light emitted by the luminophore, and repeating the sub-steps of transferring and measuring for further measurement points distributed over the surface.

Abstract: A method of correcting optical coherence tomography (OCT) image data obtained from a layered media sample includes identifying at least one interface from the obtained OCT data and correcting the OCT data for distortion at the at least one interface. The OCT image data can be corrected for extrinsic distortions, such as those caused by scan geometry, as well as, intrinsic distortions, such as those caused by refraction at each interface.

Abstract: An optical coherence tomography (OCT) system including an interferometer provides illuminating light along a first optical path to a sample and an optical delay line and collects light from the sample along a second optical path remitted at several scattering angles to a detector. In one embodiment, illuminating light is directed along a number of incident light paths through a focusing lens to a sample. The light paths and focusing lens are related to the sample and to both the incident light source and the detector. In another embodiment, a focusing system directs light to a location in the sample. A transmission grating or acousto-optic modulator directs light from the sample at an angle representative of the wavelength of the incident light on the transmission grating or acousto-optic modulator.

Abstract: A phase-referenced Doppler optical coherence tomography (OCT) system includes a low-coherence optical radiation source and a reference source co-propagated to a sample arm and a reference arm. The low-coherence and reference optical radiation reflected from the reference and arms is detected by a pair of detectors, yielding OCT and reference interferometric data output signals. The reference interferometric data output signal can be used to correct the OCT interferometric to yield velocity-indicating images that are free from defects due to sample motion and/or interferometer jitter.

Abstract: A method of correcting optical coherence tomography (OCT) image data obtained from a layered media sample includes identifying at least one interface from the obtained OCT data and correcting the OCT data for distortion at the at least one interface. The OCT image data can be corrected for extrinsic distortions, such as those caused by scan geometry, as well as, intrinsic distortions, such as those caused by refraction at each interface.

Abstract: A phase-referenced Doppler optical coherence tomography (OCT) system includes a low-coherence optical radiation source and a reference source co-propagated to a sample arm and a reference arm. The low-coherence and reference optical radiation reflected from the reference and arms is detected by a pair of detectors, yielding OCT and reference interferometric data output signals. The reference interferometric data output signal can be used to correct the OCT interferometric to yield velocity-indicating images that are free from defects due to sample motion and/or interferometer jitter.

Abstract: An optical coherence tomography (OCT) system including an interferometer provides illuminating light along a first optical path to a sample and an optical delay line and collects light from the sample along a second optical path remitted at several scattering angles to a detector. In one embodiment, illuminating light is directed along a number of incident light paths through a focusing lens to a sample. The light paths and focusing lens are related to the sample and to both the incident light source and the detector. In another embodiment, a focusing system directs light to a location in the sample. A transmission grating or acousto-optic modulator directs light from the sample at an angle representative of the wavelength of the incident light on the transmission grating or acousto-optic modulator.