Beam deflecting unit for multiple-axis examination in a microsco

Optical: systems and elements – Compound lens system – Microscope

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Details

359389, G02B 2100

Patent

active

060645180

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

1. Field of the Invention
The invention is directed to a beam deflecting unit for multiple-axis examination of specimens in a microscope which, on the one hand, enables the illumination of the specimen by one or more beams from several sides and/or observation of the specimen by one or more beams from several sides and, on the other hand, allows observation of the specimen at an angle to the illumination axis. The beam deflecting unit according to the invention can be used in available far-field raster scanning light microscopes and is situated between the microscope objective and the focal plane. It is adapted to the size ratios of the specimen and of the free working length of the objective.
2. Description of the Related Art
Technical scientific literature discloses confocal scanning light microscopes which have a resolution along the optical axis (axial resolution) and can generate images with an appreciably reduced depth of focus. A problem arises. in that the microscope objectives detect only a part of the total solid angle and accordingly detect only a portion of the light which is radiated in all directions from a point. As a result, the axial resolution is at least three-times worse than the axial resolution. Generally, the ratio is greater.
For an additionally increased resolution in the axial direction, a double-confocal scanning light microscope that was suggested in DE-OS 40 40 441 is characterized by the use of a second objective on the other side of the object plane, wherein both objectives illuminate a common object point simultaneously and/or detect the light proceeding from it. In the case of a coherent illumination of the object by the two objectives, the observation volume is reduced by interference along the optical axis. The reduction in observation volume is synonymous with an improvement in resolution.
Further, DE-OS 43 26 4731 discloses a scanning light microscope in which at least two objectives are arranged in such a way that they illuminate an object point simultaneously and/or collect the light proceeding from the object point, wherein at least two of the objectives do not lie on a common axis. In addition, as is the case in a double-confocal scanning light microscope, means for changing interference can be arranged in such a way that light passing through one of the objectives is coherently superposed at the object and/or on at least one of the light detectors with light passing through one of the other objectives at the object so that it interferes.
In the scanning light microscope known from DE-OS 43 26 473, two objectives are preferably arranged in such a way that their axes are perpendicular to one another, and one objective is used for the illumination of the specimen, while the second objective is used for the observation of the specimen. An improved resolution is achieved due to the fact that the illumination axis which is determined by the optical axis of the objective used for illumination is arranged perpendicular to the observation axis which is determined by the optical axis of the objective used for observation.
The high resolution is brought about in the following manner: The intensity distribution in the focal range is described by the illumination point spread function (PSF). The detection probability for the light proceeding from the focal range is described by the observation PSF when using a detection pinhole diaphragm. The PSF of a confocal microscope is the product of the illumination PSF and the observation PSF. The more extensive or spread out it is, the poorer the resolution of the microscope. Due to the approximately perpendicular arrangement of the illumination axis and the observation axis relative to one another, the large extension of the illumination PSF along its axis is compensated for by the small extension of the illumination PSF along this axis, so that a resolution of approximately equal quality is achieved along all three spatial axes.
According to another embodiment form of the scanning light microscope known fr

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