Method for light-sheet microscopic examination of specimens
A microscopic inspection and light sheet technology, applied in microscopes, optics, optical components, etc., can solve problems such as complex preparation, optical sheet thickness axial resolution, low light efficiency phase travel, etc.
Active Publication Date: 2020-04-17
CARL ZEISS MICROSCOPY GMBH
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Problems solved by technology
On the second hand, due to the specimen preparation and the size of the specimen chamber, the optical sheet is relatively thick and thus limited in the axial resolution that can be achieved
Third, sample preparation is complex and not compatible with standard sample preparation and standard assay fixation for fluorescence microscopy of single cells
However, this has the disadvantage that at ~10ms exposure time, the optimization of the SLM has to be done 100 times per second for the new laser wavelength
With conventional nematic SLMs, this cannot be achieved, so ferroelectric SLMs with their very low optical efficiency and limited phase travel have to be used
Method used
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[0045] figure 1 Firstly, the basic structure of the light sheet microscope is shown, which can be used for light sheet microscopic examination of samples. Here, the light sheet microscope is shown in an inverted configuration, which is only to be understood as an example in which light microscopes are also possible configurations in which the sample can be observed from above or from the side. The sample 1 is located in the sample chamber 2 and is surrounded by a liquid 3 , for example water or a nutrient solution. The sample chamber 2 has side walls and a bottom made of glass with a predetermined thickness, the thickness here being equal to, for example, the thickness of a conventional glass slide, eg 0.17 mm. The sample chamber 2 is supported on a sample table 4 which can be moved manually or by a motor in three spatial directions. The individual elements of the light sheet microscope are arranged below the sample chamber 2 , which has a transparent bottom 5 . Between the...
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A method for examining a specimen via light sheet microscopy includes selecting several illumination wavelengths for the specimen. To structure the illumination light, a predefined phase distribution is impressed on the phase-selective element and a predefined aperture structure is impressed on an aperture in the aperture plane. The phase-selective element is then illuminated in an intermediate image plane in an illumination beam path with illumination light, which is structured by the phase-selective element. The structured illumination light is imaged into an aperture plane arranged downstream of the phase-selective element. The aperture structure is adapted such that the zero orders of the structured illumination light in the aperture plane are substantially removed. The specimen is illuminated with the structured light sheet in the light sheet plane. Light emitted by the specimen is detected in a detection direction which forms an angle different from zero with the light sheet plane.
Description
technical field [0001] The present invention relates to a method for light-sheet microscopic examination of samples and mainly to the problem of analyzing samples labeled with various dyes. Background technique [0002] The research on biological samples has become more and more important in recent years. Among them, the irradiation of samples is realized by using a light sheet, and the plane of the light sheet (light sheet plane) and the optical axis of detection (detection direction) are sandwiched by a non-zero The angles intersect. Usually, the plane of the light sheet forms a non-zero, but usually not necessarily a right angle, with the detection direction (generally with respect to the optical axis of the detection objective). This research method is mainly used in fluorescence microscopy and is summarized under the term LSFM (Light Sheet Fluorescence Microscopy). Examples are the method described in DE 10257423 A1 and WO 2004 / 0535558 A1 based on the above-mentioned ...
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IPC IPC(8): G02B21/36G02B21/10G02B21/00
CPCG02B21/0032G02B21/10G02B21/367G02B21/16G06T2207/10056
Inventor 托马斯·卡尔克布莱纳海尔穆特·里佩尔特乔治·吉本摩尔根
Owner CARL ZEISS MICROSCOPY GMBH