Evaluation of signals of fluorescence scanning microscopy using a confocal laser scanning microscope

US10234667B2 · kind B2 · utility

Assignees

• Carl Zeiss Microscopy GmbH · DE
• Carl Zeiss AG · DE

Inventors

• Yauheni Novikau · Jena, DE
• Thomas Kalkbrenner · Jena, DE
• Tiemo Anhut · Jena, DE
• Daniel Schwedt · Weimar, DE
• Matthias Wald · Jena, DE

Key dates

• Filing date: May 9, 2016
• Grant date: Mar 19, 2019
• Expiry date: May 9, 2036

Classification

• Technology area (CPC G): Physics
• CPC primary: G02B21/008
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

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 p…