Abstract: A TIRFM-capable microscope including: a light source that generates/emits incoherent excitation light onto an optical path that includes a first projection lens system, a spatial filter, a second projection lens system and an objective. The TIRFM-capable microscope also includes a controller; wherein the first projection lens system projects excitation light onto the spatial filter that filters the excitation light with two-dimensional patterns, the spatial filter lies in a plane conjugate to a back focal plane of the objective which includes an objective lens that directs excitation light onto and receives fluorescent light from the sample, wherein, for a numerical aperture NAObj of the objective and a refractive index nspec of the sample NAObj>nspec, and the controller activates the spatial filter to select/generate various two-dimensional patterns and selects/adjusts the position/shape/size of the pattern such that TIRF illumination of the sample is generated.

Abstract: A TIRFM-capable microscope including: a light source that generates/emits incoherent excitation light onto an optical path that includes a first projection lens system, a spatial filter, a second projection lens system and an objective. The TIRFM-capable microscope also includes a controller; wherein the first projection lens system projects excitation light onto the spatial filter that filters the excitation light with two-dimensional patterns, the spatial filter lies in a plane conjugate to a back focal plane of the objective which includes an objective lens that directs excitation light onto and receives fluorescent light from the sample, wherein, for a numerical aperture NAObj of the objective and a refractive index nspec of the sample NAObj>nspec, and the controller activates the spatial filter to select/generate various two-dimensional patterns and selects/adjusts the position/shape/size of the pattern such that TIRF illumination of the sample is generated.

Abstract: For multi-dimensional high-resolution imaging a structure marked with fluorescence markers, fluorescence enabling light is focused to illuminate a measurement area in a sample. A partial area of the measurement area is subjected to fluorescence inhibiting light. The partial area omits a center of the measurement area in that an intensity distribution of the fluorescence inhibiting light comprises a line-shaped intensity minimum. A minimal extension of the intensity minimum in a direction through the center area is by a factor k?2 smaller than a diameter of the measurement area in said direction. Without spatial resolution, fluorescence light emitted out of the measurement area is measured for a plurality of consecutive angle positions of the intensity minimum about the center, while the measurement area, for each angle position, is subjected to the fluorescence enabling light. A value of the measured fluorescence light is assigned to the position of the center.

Abstract: For multi-dimensional high-resolution imaging a structure marked with fluorescence markers, fluorescence enabling light is focused to illuminate a measurement area in a sample. A partial area of the measurement area is subjected to fluorescence inhibiting light. The partial area omits a center of the measurement area in that an intensity distribution of the fluorescence inhibiting light comprises a line-shaped intensity minimum. A minimal extension of the intensity minimum in a direction through the center area is by a factor k?2 smaller than a diameter of the measurement area in said direction. Without spatial resolution, fluorescence light emitted out of the measurement area is measured for a plurality of consecutive angle positions of the intensity minimum about the center, while the measurement area, for each angle position, is subjected to the fluorescence enabling light. A value of the measured fluorescence light is assigned to the position of the center.