Method and configuration for the optical detection of an illuminated specimen

Abstract

A method and a configuration for the depth-resolved optical detection of a specimen, in which a specimen or a part of the specimen is scanned by means of preferably linear illumination. The illumination of the specimen is periodically structured in the focus in at least one spatial direction. Light coming from the specimen is detected and images of the specimen are generated. At least one optical sectional image and/or one image with enhanced resolution is calculated through the specimen. Images are repeatedly acquired and sectional images are repeatedly blended while changing the orientation of the linear illumination relative to the specimen and/or spatial intervals between lines exposed to detection light from the illuminated specimen region are generated for the line-by-line non-descanned detection on an area detector or a camera and/or, during a scan, light is further deflected upstream of the detector through the line in the direction of the scan of the specimen.

1 . A method for the depth-resolved optical detection of a specimen, wherein a specimen or a part of the specimen is scanned by means of linear illumination,
 the illumination of the specimen being periodically structured in the focus in at least one spatial direction,   light coming from the specimen being detected and images of the specimen being generated,   and at least one optical sectional image and/or one image with enhanced resolution being calculated through the specimen determined,   comprising the steps of   repeatedly acquiring images, and repeatedly blending sectional images while changing the orientation of the linear illumination relative to said specimen.   
     
     
         2 . The method of  claim 1 , wherein the line is rotated about the optical axis, and the images are generated and the sectional images are blended at different angles of rotation. 
     
     
         3 . The method of  claim 1 , wherein, to generate lines, a beam-shaping unit is jointly rotated with means for structuring the illuminating light. 
     
     
         4 . A method for the depth-resolved optical detection of a specimen, wherein a specimen or a part of the specimen is scanned by means of preferably linear illumination,
 the illumination of the specimen being periodically structured in the focus in at least one spatial direction,   light coming from the specimen being detected and images of the specimen being generated,   and at least one optical sectional image and/or one image with enhanced resolution being calculated through the specimen is determined, comprising the steps of   generating spatial intervals between lines exposed to detection light from the illuminated specimen region for the line-by-line non-descanned detection on an area detector or a camera.   
     
     
         5 . The method of  claim 4 , wherein during the preferably linear illumination and detection, the illumination is repeatedly switched on and off. 
     
     
         6 . The method of  claim 4 , wherein during the scanning of the specimen, the light is repeatedly interrupted so that a spatial interval is formed between two illuminated specimen regions. 
     
     
         7 . The method of  claim 4  for the confocal generation of images, wherein an image is calculated by partially or completely masking the spatial intervals between camera regions associated with the exposed specimen regions and the images thus obtained are blended. 
     
     
         8 . The method of  claim 7 , wherein the images are blended in such a manner that neighboring scanned regions of the specimen are properly scaled and adjacently aligned in the blended image. 
     
     
         9 . A method for the depth-resolved optical detection of a specimen, wherein a specimen or a part of a specimen is scanned by means of preferably linear illumination,
 the illumination of the specimen being periodically structured in the focus in at least one spatial direction,   light coming from the specimen being detected and images of the specimen being generated,   and at least one optical sectional image and/or one image with enhanced resolution being calculated through the specimen is determined, wherein, during a scanning procedure, light is further deflected upstream of the detector through the line in the direction of the scan of the specimen.   
     
     
         10 . The method of  claim 9 , wherein the speed of the light deflection is greater than the speed of the relative movement between the specimen and the illuminating light. 
     
     
         11 . The method of  claim 9 , wherein the light is deflected step-by-step. 
     
     
         12 . The method of  claim 9 , wherein the light is deflected continuously. 
     
     
         13 . The method of  claim 9 , wherein, during a rotation of the illuminating line, the polarization of the illuminating light is synchronously rotated with the rotation. 
     
     
         14 . The method of  claim 9 , wherein repeated scanning takes place and the position of the periodic structure on the specimen and/or the position of the illuminating light on the specimen is shifted. 
     
     
         15 . The method of  claim 9 , wherein several images with different image phases are acquired and sectional images are calculated therefrom. 
     
     
         16 . The method of  claim 9 , wherein the images are acquired with different image phases with a constant spatial interval between illuminated/detected sections. 
     
     
         17 . The method of  claim 9 , wherein the position of the spatial interval is changed and several images with different image phases are acquired for each position and sectional images are calculated therefrom. 
     
     
         18 . The method of  claim 9 , wherein first the position of the spatial interval for one position of the structure is changed and specimen images are acquired, and subsequently this procedure is repeated for the next position of the structure. 
     
     
         19 . The method of  claim 9 , wherein the position of the spatial interval is changed in such a manner that substantially all specimen regions are sequentially illuminated line-by-line and the specimen light is detected. 
     
     
         20 . The method of  claim 9 , wherein the light is interrupted by decreasing the intensity by means of an electro-optical and/or acousto-optical modulator. 
     
     
         21 . The method of  claim 9 , wherein, for the purpose of the periodic structuring of the illumination, a light beam is divided into several component light beams, which light beams are interferometrically overlapped and shaped into a line. 
     
     
         22 . The method of  claim 9 , wherein the light resulting from a nonlinear interaction of the illumination with the specimen or a part of the specimen . . . [word or words missing] and is detected. 
     
     
         23 . The method of  claim 9 , wherein linear scanning is carried out simultaneously with several lines. 
     
     
         24 . The method of  claim 9 , wherein the optical section thickness or optical resolution is varied as structures with different modulation frequencies are imaged. 
     
     
         25 . The method of  claim 9 , wherein during illumination with several wavelengths, the section thickness is identically set by adjusting each modulation frequency. 
     
     
         26 . A configuration for the depth-resolved optical detection of a specimen, comprising
 means for the preferably linear illumination of the specimen with at least one wavelength,   means for spatially structuring the illuminating light in at least one plane,   means for generating a relative movement between the specimen and the illuminating light,   means for imaging the light influenced by the specimen on at least one detector,   means for calculating at least one optical sectional image and/or one image with enhanced resolution from the spatial information of the light influenced by the specimen, and means for changing the orientation of the linear illumination relative to the specimen.   
     
     
         27 . The configuration of  claim 26 , further comprising a jointly rotatable unit comprising a beam-shaping unit to generate lines and means for structuring the illuminating light in the beam path. 
     
     
         28 . A configuration for the depth-resolved optical detection of a specimen, comprising
 means for the preferably linear illumination of the specimen with at least one wavelength,   means for spatially structuring the illuminating light in at least one plane,   means for generating a relative movement between the specimen and the illuminating light,   means for imaging the light influenced by the specimen on at least one detector,   means for calculating at least one optical sectional image and/or one image with enhanced resolution from the spatial information of the light influenced by the specimen,   an area detector or a camera for the non-descanned detection of the specimen light, and   means for interrupting the light during the scan so as to generate a spatial interval between illuminated specimen regions and/or to generate spatial intervals between lines exposed with detection line from the illuminated specimen region on the area detector.   
     
     
         29 . The configuration of  claim 28 , wherein intensity control means are disposed in the illuminating beam path. 
     
     
         30 . The configuration of  claim 28  or  29 , wherein an electro- or acousto-optical modulator for light interruption is provided. 
     
     
         31 . A configuration for the depth-resolved optical detection of a specimen, comprising
 means for the preferably linear illumination of the specimen with at least one wavelength,   means for spatially structuring the illuminating light in at least one plane,   means for generating a relative movement between the specimen and the illuminating light,   means for aging the light influenced by the specimen on at least one detector,   means for calculating at least one optical sectional image and/or one image with enhanced resolution from the spatial information of the light influenced by the specimen, and   a scanner disposed in the detection beam path in order to expand the specimen light discretely on the detector or continuously on the detector during the line-by-line scan.   
     
     
         32 . The configuration of  claim 31 , characterized in that at least one scanner is provided as a means for generating the relative movement. 
     
     
         33 . The configuration of  claim 31 , wherein the means for structuring the illumination is an optical element that is preferably rotatable about the optical axis and that is structured relative to its transparency. 
     
     
         34 . The configuration of  claim 31 , wherein in order to set different image phases of the structure, the position of at least one scanner can be adjusted. 
     
     
         35 . The configuration of  claim 31 , wherein in order to set different frequency structures, gratings of different periodicities that can be rotated into the beam path are provided. 
     
     
         36 . A microscope, preferably a laser scanning microscope, having the configuration of  claim 26 ,  28  or  31 .