# US 2022/0034813 A1 · Optical arrangment for fluorescence microscopy applications > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2022/0034813 A1 - Document ID: US20220034813A1 - Status: Published application - Kind: A1 - Application number: 17/275,705 - Filed: 2019-09-12 - Published: 2022-02-03 - Earliest claimed: 2018-09-18 - Assignees: Fraunhofer-Gesellschaft Zur Foerderung Angewandten Forschung E.V.; Friedrich-Schiller-Universitaet Jena - Applicants: Fraunhofer-Gesellschaft Zur FöRderung Der Angewandten Forschung E.V.; Friedrich-Schiller-Universitaet Jena - Inventors: Markus GRAEFE; Marta GILABERTE BASSET; Falk EILENBERGER; Frank SETZPFAND - Web page: https://patentgrep.com/patent/US20220034813A1 ## CPC - G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)} - G02B 21/0032: {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)} - G02B 21/0076: {arrangements using fluorescence or luminescence} - G02B 21/06: Means for illuminating specimens - G02B 21/16: adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)} - G02B 21/367: {providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison} - G02B 2207/114: Two photon or multiphoton effect - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS ## Abstract In the optical arrangement for fluorescent microscopic applications, one or more multiphoton beams, but at least one or two photon pair beams, from a source of non-classical light is/are directed at a first optical system, consisting of an arrangement of at least one lens or one photon-reflecting element or another beam-forming element or a combination thereof. The first optical system (3) is designed to shape the non-classical light into a light sheet (4) or a light sheet-like shape and thence to direct it at a specimen (5), so that fluorescent radiation is excited by means of multiphoton absorption using the multiple multiphoton beams that are simultaneously incident on/in the specimen. Fluorescent radiation (6) obtained by excitation is incident by means of a second optical system (7) on a detection system (8) that is designed for the spatially resolved capture of fluorescent radiation. ## Claims ### Claim 1 Optical arrangement for fluorescence microscopy applications, in which from a source of non-classical light one or more multiphoton beam(s), but at least one or two photon pair beam(s) is/are directed onto a first optical system consisting of an arrangement of at least one lens or photon reflecting element or other beam shaping element or a combination thereof, the first optical system is adapted to form the non-classical light into a light sheet or a light sheet-like shape, directed to a sample such that fluorescence radiation is excited with several multiphoton beams incident simultaneously on/in the sample by means of multiphoton absorption, and fluorescence radiation obtained by excitation occurs, by means of a second optical system, on a detection system which is designed for spatially resolved detection of fluorescence radiation. ### Claim 2 (depends on 1) The arrangement according to claim 1, wherein the source of nonclassical light is a nonlinear crystal pumped by a laser or waveguide structure in a nonlinear crystal, or at least two identical coherently pumped quantum dots. ### Claim 3 (depends on 1) The arrangement according to claim 1, wherein the one or more multiphoton beam(s), but at least the one or more photon pair beam(s), is/are directed, in collinear geometry, towards the first optical system. ### Claim 4 (depends on 1) The arrangement according to claim 1, wherein the fluorescence radiation can be excited with two photons in the form of a photon pair. ### Claim 5 (depends on 1) The arrangement according to claim 1, wherein the formation of the light sheet is performed by means of the first optical system, which is adapted to linearly change at least one multiphoton beam the position of impingement of the at least one multiphoton beam on/in the sample by a movement of an optical element which is part of the first optical system, so that a corresponding line-shaped region of at least one line is irradiated at least once. ### Claim 6 (depends on 1) The arrangement according to claim 1, wherein a multiphoton beam emitted by a photon beam source is split into a plurality of partial beams and the partial beams are directed onto/into the sample by means of the first optical system for forming a respective light sheet. ### Claim 7 (depends on 1) The arrangement according to claim 1, wherein the formation of at least one light sheet or light sheet-like shape is effected by means of the first optical system which is designed to linearly change a position of impingement of the partial beams on/in the sample by a movement of at least one optical element which is a component of the first optical system, so that a corresponding line-shaped region of at least one line is irradiated at least once with a partial beam. ### Claim 8 (depends on 1) The arrangement according to claim 1, wherein a one- or two- or three-dimensional movement of the sample is performed for spatially resolved imaging of the sample. ### Claim 9 (depends on 1) The arrangement according to claim 1, wherein the first optical system or the second optical system is formed including a nonlinear optical crystal, an optical lens, a photon reflecting element, a polarization optics, an optical filter or an arrangement of a plurality of these optical elements. ### Claim 10 (depends on 1) An arrangement according to claim 1, wherein the multiphoton beam emitted collinearly by the source of non-classical light is divisible by means of a dichroic mirror or a polarization beam splitter into a plurality of partial beams directed at/into the sample at different positions. ### Claim 11 (depends on 1) The arrangement according to claim 1, wherein the detector system is a CCD, an EMCCD, an ICCD or a CMOS camera or a SPAD array.