Microscope and method for SPIM microscopy

Patent metadata

Field	Value
Publication number	US-10698225-B2
Application number	US-201314411710-A
Country	US
Kind code	B2
Filing date	Jun 26, 2013
Priority date	Jul 2, 2012
Publication date	Jun 30, 2020
Grant date	Jun 30, 2020

Abstract

A microscope, in particular according to any of the preceding claims, consisting of an illuminating device, comprising an illumination light source and an illumination beam path for illuminating the specimen with a light sheet, a detection device for detecting light emitted by the specimen, and an imaging optical unit, which images the specimen via an imaging objective in an imaging beam path at least partly onto the detection device, wherein the light sheet is essentially planar at the focus of the imaging objective or in a defined plane in proximity of the geometrical focus of the imaging objective, and wherein the imaging objective has an optical axis, which intersects the plane of the light sheet at an angle that is different from zero, preferably perpendicularly, wherein an amplitude and/or phase filter is provided in the illumination beam path, said filter acting as a sine spatial filter in that it limits the illumination light in at least one spatial direction by filtering the spatial frequencies that occur with a sine function and/or in that the illumination light is limited with regard to the phase and amplitude thereof in at least one spatial direction by a sine filter function, and/or a combined amplitude and phase filter is provided in the illumination beam path, said filter shaping the light sheet by influencing the transmission profile of the illumination light distribution with a sine filter function.

First claim

What is claimed is:

1. A microscope comprising:
an illuminating device, having an illumination light source and an illumination beam path with an optical axis for illuminating a specimen with a light sheet,
a detection device for detecting light emitted by the specimen,
an imaging optical unit, which images the specimen via an imaging objective in an imaging beam path at least partly onto the detection device,
said light sheet being planar at the focus of the imaging objective or in a defined plane in proximity of the geometrical focus of the imaging objective, and said imaging objective having an optical axis, which intersects the plane of the light sheet at an angle that is different from zero,
an amplitude or phase filter provided in the illumination beam path, said filter acting as a sinc(z) spatial filter by limiting the illumination light in at least a spatial direction z parallel to said optical axis of said illumination beam path by filtering with a sinc function the occurring spatial frequencies and enabling a constant beam profile on the axis across an area of the light sheet in the z direction.

2. A microscope according to claim 1 , wherein said amplitude or phase filter is arranged at least partly in or in proximity of a plane of the aperture diaphragm of an illumination optical unit and/or the filter is realized with a hologram (CGH) and/or a diffractive optical unit and/or the filter is formed at least partly by a spatial light modulator (SLM) and/or the light sheet is displaced perpendicular to the direction of the light with the SLM and/or the focal plane of the imaging objective is displaced when the light sheet is displaced and/or that an asymmetrical sinc function/off-axis distribution is generated in the plane of the aperture diaphragm and/or that a sinc filter acting extra-axially to the optical axis is provided and/or a superimposition of a plurality of sinc filters is provided for structuring the light sheet and/or a sinc function rescaled with the Ewald summation is provided as filter function and/or the filter is combined with an anamorphic optical unit in the illumination beam path for forming the beam.

3. A microscope according to claim 1 , wherein said angle that is different than zero being ninety degrees.

Assignees

• Zeiss Carl Microscopy Gmbh

Inventors

• Singer Wolfgang
• Siebenmorgen Jorg
• Anhut Tiemo
• Wolleschensky Ralf

Classifications

• G02B21/0032 Primary

  Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers (G02B21/0036 - G02B21/008; means for illumination of specimens in general G02B21/06) · CPC title

• G02B21/0076

  arrangements using fluorescence or luminescence · CPC title

• G02B21/367

  providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison · CPC title

• G02B21/06

  Means for illuminating specimens · CPC title

• G02B27/0927 Primary

  Systems for changing the beam intensity distribution, e.g. Gaussian to top-hat · CPC title