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Method for adjusting a laser-scanning fluorescence microscope and laser-scanning fluorescence microscope having an automatic adjustment device

A fluorescence microscope, laser scanning technology, used in microscopes, fluorescence/phosphorescence, optics, etc., can solve problems such as not allowing alignment

Active Publication Date: 2019-06-21
ABBERIOR INSTR GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In other words, the calibration specimen only allows alignment in the range of the intermediate image, but does not allow alignment in the sample at the range of the focal point of the lens

Method used

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  • Method for adjusting a laser-scanning fluorescence microscope and laser-scanning fluorescence microscope having an automatic adjustment device
  • Method for adjusting a laser-scanning fluorescence microscope and laser-scanning fluorescence microscope having an automatic adjustment device
  • Method for adjusting a laser-scanning fluorescence microscope and laser-scanning fluorescence microscope having an automatic adjustment device

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Experimental program
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Embodiment Construction

[0061] exist figure 1 The laser scanning fluorescence microscope 15 shown in has an excitation light source 16 for excitation light 6 . The excitation light 6 is reflected by the beam splitter 26 to another beam splitter 25 . The beam splitter 25 is used to superimpose the excitation light 6 with the fluorescence blocking light 7 which comes from the fluorescence blocking light source 17 . In this case, a wavefront modulator 24 is arranged in the beam path of the fluorescence-prevented light 7 between the fluorescence-prevented light source 17 and the beam splitter 25 . The excitation light 6 combined with the fluorescence blocking light 7 is directed through the beam splitter 25 , through the scanner 27 and through the focusing lens 20 into the sample space 21 . A sample 22 is arranged in the sample space 21 , into which the excitation light 6 and the fluorescence prevention light 7 are focused. Fluorescent light 19 emitted by sample 22 passes through lens 20, scanner 27 a...

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Abstract

The invention relates to a method for setting a correct adjustment of a laser scanning fluorescence microscope (15), in which an intensity maximum (1) of stimulation light (6) and an intensity maximum(4) of fluorescence prevention light (7) and / or an imaging of a pinhole aperture (2) arranged in front of a fluorescence light detector (18) coincide in the focus of a lens (20), wherein a structuremarked by a fluorescence dye is scanned in a sample (22) at the intensity maximum (1) of the stimulation light (6), in order to produce images (A, B) of the sample (22) having maps (3, 30, 31, 32) ofthe structure, and, from the maps (3, 30, 31, 32) of the structure in the images, an offset (8) between positions of the maps of the structure in the generated images is calculated and balanced. The generated images correspond to different intensities of the fluorescence prevention light (7) and / or to different aperture openings of the pinhole aperture (2).

Description

technical field [0001] The invention relates to a method for adjusting the correct alignment of a laser scanning fluorescence microscope in which the maximum intensity of the excitation light and the minimum intensity of the fluorescence blocking light and / or the diaphragm arranged in front of the fluorescence detector The images (Abbildung) of are coincident in the focal point of the lens. In particular, the present invention relates to a method that uses a metric (Metrik) for setting the alignment, which is calculated and enables a precise automatic alignment. [0002] Furthermore, the invention relates to a laser scanning fluorescence microscope with an automatic alignment device for carrying out such a method. [0003] A laser scanning fluorescence microscope is in particular a confocal microscope with a diaphragm arranged in front of the fluorescence detector and / or a high-resolution laser scanning fluorescence microscope, in which case a laser scanning fluorescence micr...

Claims

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Application Information

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IPC IPC(8): G02B21/00G01N21/63G01N21/64
CPCG01N21/636G01N21/6458G02B21/0072G02B21/0076G02B21/008
Inventor J·海涅M·罗伊斯B·哈尔克L·卡斯特鲁普
Owner ABBERIOR INSTR GMBH