Abstract: For spatial high resolution determining a position of a singularized molecule, which is excitable with excitation light for emission of luminescence light, in a sample, the excitation light is provided with an intensity distribution comprising an intensity increasing region with a known strictly monotonic course of an intensity of the luminescence light over a distance of the singularized molecule to a model point of the intensity distribution. The model point is arranged at different preliminary positions such that the intensity increasing region extends over a preliminary local area of the sample including the singularized molecule. From intensity values including intensities of the luminescence light separately registered for the preliminary positions of the model point, a further local area is determined which includes the singularized molecule and which is smaller than the preliminary local area. These steps are repeated using the last further local area as the next preliminary local area.

Abstract: For spatial high resolution determining a position of a singularized molecule, which is excitable with excitation light for emission of luminescence light, in n spatial dimensions in a sample, the excitation light is directed onto the sample with an intensity distribution, which has a zero point and intensity increasing regions adjoining the zero point on both sides in each of the n spatial dimensions. The zero point is arranged at not more than n×3 different positions. The luminescence light emitted by the singularized molecule is separately registering for each of the different positions of the zero point. The position of the singularized molecule in the n spatial dimensions in the sample is deduced from intensities of the luminescence light separately registered for the not more than n×3 different positions of 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: A method of spatially measuring a plurality of nano-scale structures in a sample comprises the steps of: marking the individual structures at different locations with fluorescent markers, coupling the individual structures to individual positioning aids whose positions in the sample are known, exciting the fluorescent markers with excitation light for emission of fluorescence light, wherein an intensity distribution of the excitation light has a local minimum, arranging the local minimum at different positions in a close-up range around the position of respective positioning aid whose dimensions are not larger than the diffraction limit at the wavelength of the excitation light, registering the fluorescence light emitted out of the sample separately for the individual fluorescent markers and for the different positions of the minimum, and determining positions of the individual fluorescent markers in the sample from the intensities of the fluorescence light registered.

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: A scanning luminescence light microscope for spatial high resolution imaging a structure marked with a luminescent marker comprises a light source for luminescence inhibition light and for further light; a light shaping and aligning device; and a detector registering luminescence light emitted by the luminescent marker. The device, by means of two optical gratings and an objective lens, forms two crossing line gratings of the luminescence inhibition light, and two crossing line gratings of the further light so that local intensity minima of an overall intensity distribution of the luminescence inhibition light are delimited in at least two directions, and that local intensity maxima or local intensity minima of an overall intensity distribution of the further light coincide with the local intensity minima of the luminescence inhibition light. Further, the device moves the overall intensity distributions of the further light and the luminescence inhibition light to scan the structure.

Abstract: In order to determine the locations of individual fluorescent molecules in a sample, which keep a minimum distance with regard to each other, the individual molecules are excited for emission of fluorescence light by means of excitation light. The fluorescence light is registered for different positions of a zero point of an intensity distribution of the excitation light. The distance between these positions is at least half the minimum distance of the fluorescent molecules. The locations of the fluorescent molecules are derived from the course of the intensity of the fluorescence light over the positions of the zero point of the excitation light.

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: The invention relates to novel fluorescent dyes with phosphorylated hydroxymethyl groups, a method for preparing the same as well as to their use in imaging techniques. The fluorescent dyes are coumarin, rhodamine or BODIPY dyes having of one of the following general formulae I-III: wherein W=OP(O)Y1Y2 or P(O)Y1Y2, where Y1 and Y2 independently denote any of the following residues: OH, O(?), ORa and ORb, NHRa and NHRb, NRaRb and NRcRd, ORa and NHRb, ORa and NRbRc, NHRa and NRbRc; and any salt thereof.

Abstract: A scanning luminescence light microscope for spatial high resolution imaging a structure marked with a luminescent marker comprises a light source for luminescence inhibition light and for further light; a light shaping and aligning device; and a detector registering luminescence light emitted by the luminescent marker. The device, by means of two optical gratings and an objective lens, forms two crossing line gratings of the luminescence inhibition light, and two crossing line gratings of the further light so that local intensity minima of an overall intensity distribution of the luminescence inhibition light are delimited in at least two directions, and that local intensity maxima or local intensity minima of an overall intensity distribution of the further light coincide with the local intensity minima of the luminescence inhibition light. Further, the device moves the overall intensity distributions of the further light and the luminescence inhibition light to scan the structure.

Abstract: In order to determine the locations of individual fluorescent molecules in a sample, which keep a minimum distance with regard to each other, the individual molecules are excited for emission of fluorescence light by means of excitation light. The fluorescence light is registered for different positions of a zero point of an intensity distribution of the excitation light. The distance between these positions is at least half the minimum distance of the fluorescent molecules. The locations of the fluorescent molecules are derived from the course of the intensity of the fluorescence light over the positions of the zero point of the excitation light.

Abstract: For spatial high resolution imaging of a structure of a sample, the structure comprising a luminophore, the sample, in a measurement area, is subjected to an intensity distribution of luminescence inhibiting light comprising a local minimum. Then, the sample, in the measurement area, is subjected to luminescence excitation light which excites the luminophore out of an electronic ground state into a luminescent state, and luminescence light emitted out of the measurement area is registered. This registered luminescence light is assigned to the position of the local minimum within the sample. The luminescence inhibiting light disturbs the electronic ground state of the luminophore such that the luminophore, in the disturbed electronic ground state, has an absorption cross-section for the luminescence excitation light which is reduced by at least 50% as compared to the undisturbed electronic ground state.

Abstract: For the purpose of tracking a movement of a particle in a sample, the particle is driven by light to emit photons, and the photons emitted by the particle are detected. The light applied to the sample features a light intensity distribution with a spatially limited minimum. The particle is tracked with the minimum of the light intensity distribution by moving the light intensity distribution with respect to the sample such that a rate of photons emitted by the particle remains minimal, and by taking an actual position of the minimum of the light intensity distribution in the sample as an actual position of the particle in the sample.

Abstract: For spatial high resolution imaging of a structure of a sample comprising a luminophore, the sample is subjected to excitation inhibiting light transferring the luminophore out of an excitable electronic ground state into a protection state in which the luminophore is protected against electronic excitation by luminescence excitation light and luminescence de-excitation light. The excitation inhibiting light comprises a first local minimum. Next, the sample is subjected to the luminescence excitation light exciting the luminophore within the first local minimum into an excited luminescent state. Then, the sample is subjected to the luminescence de-excitation light returning the luminophore out of the excited luminescent state into the excitable electronic ground state. The luminescence de-excitation light comprises a second local minimum overlapping with the first local minimum.

Abstract: The invention relates to novel fluorescent dyes with phosphorylated hydroxymethyl groups, a method for preparing the same as well as to their use in imaging techniques. The fluorescent dyes are coumarin, rhodamine or BODIPY dyes having of one of the following general formulae I-III: wherein W=OP(O)Y1Y2 or P(O)Y1Y2, where Y1 and Y2 independently denote any of the following residues: OH, O(?), ORa and ORb, NHRa and NHRb, NRaRb and NRcRd, ORa and NHRb, ORa and NRbRc, NHRa and NRbRc; and any salt thereof.

Abstract: For spatial high resolution imaging of a structure of a sample, the structure comprising a luminophore, the sample, in a measurement area, is subjected to an intensity distribution of luminescence inhibiting light comprising a local minimum. Then, the sample, in the measurement area, is subjected to luminescence excitation light which excites the luminophore out of an electronic ground state into a luminescent state, and luminescence light emitted out of the measurement area is registered. This registered luminescence light is assigned to the position of the local minimum within the sample. The luminescence inhibiting light disturbs the electronic ground state of the luminophore such that the luminophore, in the disturbed electronic ground state, has an absorption cross-section for the luminescence excitation light which is reduced by at least 50% as compared to the undisturbed electronic ground state.

Abstract: For spatial high resolution imaging of a structure of a sample comprising a luminophore, the sample is subjected to excitation inhibiting light transferring the luminophore out of an excitable electronic ground state into a protection state in which the luminophore is protected against electronic excitation by luminescence excitation light and luminescence de-excitation light. The excitation inhibiting light comprises a first local minimum. Next, the sample is subjected to the luminescence excitation light exciting the luminophore within the first local minimum into an excited luminescent state. Then, the sample is subjected to the luminescence de-excitation light returning the luminophore out of the excited luminescent state into the excitable electronic ground state. The luminescence de-excitation light comprises a second local minimum overlapping with the first local minimum.

Abstract: The invention relates to novel fluorescent dyes with phosphorylated hydroxymethyl groups, a method for preparing the same as well as to their use in imaging techniques. The fluorescent dyes are coumarin, rhodamine or BODIPY dyes having of one of the following general formulae I-III: wherein W=OP(O)Y1Y2 or P(O)Y1Y2, where Y1 and Y2 independently denote any of the following residues: OH, O(?), ORa and ORb, NHRa and NHRb, NRaRb and NRcRd, ORa and NHRb, ORa and NRbRc, NHRa and NRbRc; and any salt thereof.

Abstract: For determining the distribution of a substance, a measuring front is formed of a first and a second optical signal. Intensities of the first and second optical signals, over a depth of the measuring front which is smaller than the diffraction limit at the wavelengths of the first and second optical signals, increase so steeply that a portion of the substance in a measurement state in which a measurement signal is available from the substance increases from essentially zero due to transferring the substance by means of the first optical signal into the measurement state, and decreases to essentially zero again due to transferring the substance by means of the second optical signal back out of the measurement state. The measuring front is moved over a measurement region. The measurement signal is recorded for different positions of the measuring front in the measurement region and assigned to these positions.