Method and Configuration for the Optical Detection of an Illuminated Specimen

Abstract

A method for the optical detection of an illuminated specimen, wherein the illuminating light impinges in a spatially structured manner in at least one plane on the specimen and several images of the specimen are acquired by a detector in different positions of the structure on the specimen. An optical sectional image and/or an image with enhanced resolution is then calculated. The method includes generating a diffraction pattern in the direction of the specimen in or near the pupil of the objective lens or in a plane conjugate to the pupil. A phase plate with regions of varying phase delays is dedicated to the diffraction pattern in or near the pupil of the objective lens or in a plane conjugate to said pupil, and different phase angles of the illuminating light are set.

1 . A configuration for the optical detection of an illuminated specimen, comprising illuminating light having a light path, and a microscope objective lens, wherein illuminating light impinges in a spatially structured manner in at least one plane on said specimen, thereby creating a spatially periodic light structure in the specimen, and several images of the specimen are acquired by a detector in different positions of the light structure on the specimen, from which images an optical sectional image and/or an image with enhanced resolution is calculated, an illuminating-light distribution generated in or near the pupil of said objective lens or in a plane conjugate to said pupil, said light distribution being formed by at least two light spots, and a movable diaphragm for selecting two of said light spots, said diaphragm being disposed in or near the pupil of said objective lens or in a plane conjugate to said pupil, wherein said periodic light structure is formed by said objective in the specimen from said selected light spots and wherein said different positions of the light structure are created by selecting different sets of said light spots via said diaphragm. 
     
     
         2 . The configuration of  claim 1 , wherein a spatially periodic structure for creating said light spots is disposed in an intermediate image plane. 
     
     
         3 . The configuration of  claim 1 , wherein the structure has an N-fold symmetry, N=2, 4, 6 . . . , which, in or near the pupil, generates said light spots for structuring in N/2 spatial directions within the specimen, which illumination patterns are symmetrical relative to the optical axis. 
     
     
         4 . The configuration of  claim 1 , further comprising a phase plate located in the path of the illuminating light. 
     
     
         5 . The configuration of  claim 4 , wherein the phase plate is circular with regions of different phase delays, which regions have at least partially the shape of circular sectors, and wherein said phase plate can rotate about the optical axis. 
     
     
         6 . The configuration of  claim 4 , wherein the different phase delays are produced by different refractive indices and/or different plate thicknesses. 
     
     
         7 . The configuration of  claim 4 , wherein the phase delay for chromatic correction varies perpendicular to the optical axis. 
     
     
         8 . The configuration of  claim 4 , wherein said phase plate can be continuously rotated or moved and is synchronized with the image acquisition, and wherein an image is acquired once a region with a phase delay different from the preceding one is reached. 
     
     
         9 . The configuration of  claim 4 , wherein said phase plate can be rotated or moved in discrete steps. 
     
     
         10 . The configuration of  claim 1 , wherein said diaphragm is a circular mask with oppositely lying transmission openings, which mask can rotate about the optical axis. 
     
     
         11 . The configuration of  claim 4 , wherein said diaphragm and said phase plate are disposed on two rotatable or movable elements that are connected to each other. 
     
     
         12 . The configuration as in  claim 11 , wherein the connection is implemented via a gear unit. 
     
     
         13 . The configuration of  claim 11  wherein said diaphragm and/or phase plate is/are driven by a stepping motor. 
     
     
         14 . The configuration of  claim 11 , wherein the light distribution is produced by outputs of fiber optics. 
     
     
         15 . A method for the optical detection of an illuminated specimen using illuminating light having a light path and a microscope objective lens, wherein the illuminating light impinges in a spatially structured manner in at least one plane on the specimen, thereby creating a spatially periodic light structure in the specimen, several images of the specimen are acquired by a detector in different positions of the light structure on the specimen, from which images an optical sectional image and/or an image with enhanced resolution is calculated, said method comprising: generating an illuminating-light distribution in or near the pupil of said objective lens or in a plane conjugate to said pupil; said light distribution being formed by at least two light spots; selecting two of said light spots via a movable diaphragm disposed in or near the pupil of said objective lens or in a plane conjugate to said pupil, wherein said periodic light structure is formed by said objective in the specimen from said selected light spots and wherein said different positions of the light structure are created by selecting different sets of said light spots via said diaphragm. 
     
     
         16 . The method of  claim 15 , wherein said light spots are created by providing a spatial periodic structure in an intermediate image plane. 
     
     
         17 . The method of  claim 16 , wherein said structure has an N-fold symmetry, N=2, 4, 6 . . . , which, in or near the pupil, generates said light spots, symmetrical relative to the optical axis, for a structure in N/2 spatial directions within the specimen. 
     
     
         18 . The method of  claim 15 , further comprising providing a phase plate in the path of the illuminating light. 
     
     
         19 . The method of  claim 18 , wherein said phase plate has a circular shape with regions with different phase delays, which regions have an at least partially circular shape, and can be rotated about the optical axis. 
     
     
         20 . The method of  claim 18 , wherein a different phase delay is set by means of different refractive indices and different plate thicknesses. 
     
     
         21 . The method of  claim 18 , wherein said phase delay for the purpose of chromatic correction is varied perpendicular to the optical axis. 
     
     
         22 . The method of  claim 18 , wherein said phase plate is continuously rotated or moved and synchronized with the image acquisition, and wherein an image is acquired once a region with a phase delay different from the preceding one is reached. 
     
     
         23 . The method of  claim 18 , wherein said phase plate is rotated/moved in discrete steps. 
     
     
         24 . The method of  claim 15 , wherein said light spots are selected by way of said movable diaphragm which is disposed in or near the pupil of the objective lens or in a plane conjugate to said pupil. 
     
     
         25 . The method of  claim 24 , wherein said diaphragm is a circular mask with oppositely lying transmission openings and wherein said diaphragm rotates about the optical axis. 
     
     
         26 . The method of  claim 18 , wherein said diaphragm and said phase plate are disposed on two rotatable/movable elements that are connected to each other. 
     
     
         27 . The method of  claim 15 , wherein the light distribution is generated by outputs of fiber optics.