- 专利标题: Evaluation of signals of fluorescence scanning microscopy using a confocal laser scanning microscope
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申请号: US15573429申请日: 2016-05-09
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公开(公告)号: US10234667B2公开(公告)日: 2019-03-19
- 发明人: Yauheni Novikau , Thomas Kalkbrenner , Tiemo Anhut , Daniel Schwedt , Matthias Wald
- 申请人: Carl Zeiss Microscopy GmbH , Carl Zeiss AG
- 申请人地址: DE Jena DE Oberkochen
- 专利权人: Carl Zeiss Microscopy GmbH,Carl Zeiss AG
- 当前专利权人: Carl Zeiss Microscopy GmbH,Carl Zeiss AG
- 当前专利权人地址: DE Jena DE Oberkochen
- 代理机构: Duane Morris LLP
- 优先权: DE102015107367 20150511
- 国际申请: PCT/DE2016/100207 WO 20160509
- 国际公布: WO2016/180403 WO 20161117
- 主分类号: G02B21/00
- IPC分类号: G02B21/00 ; G01N21/64
摘要:
A method for evaluating signals of fluorescence scanning microscopy with simultaneous excitation and detection of fluorescence in different focal planes of a sample by means of confocal laser scanning microscopy. The invention evaluates signals of fluorescence scanning microscopy without the signal losses usually taking place with a confocal aperture, by coupling an illumination beam into a microscope observation beam path which images a measuring volume on a detector array arranged in the image plane, focusing the illumination beam which passes through a beam-forming phase mask for generating an elongated focus in the measuring volume, collecting and collimating fluorescent light generated in the measuring volume and routing it to diffractive optics which split the light beams into different diffraction orders and impress a different spherical phase on the light beams, imaging the different diffraction orders on detector regions of the detector array so that fluorescent light from focal planes at different depths of the measuring volume are associated with different diffraction orders, and associating the fluorescence signals on which crosstalk is superposed from different focal planes of the measuring volume with defined focal planes by means of correlation-based association based on distinguishable blinking behavior of fluorescing dyes.
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