# US 12,216,056 B2 · Device for wavefront analysis and microscopic imaging systems comprising such analysis devices > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,216,056 B2 - Document ID: US12216056B2 - Status: Granted - Kind: B2 - Application number: 18/606,830 - Filed: 2024-03-15 - Published: 2024-07-11 - Granted: 2025-02-04 - Earliest claimed: 2019-02-01 - Assignees: Imagine Optic - Applicants: Imagine Optic - Inventors: Fabrice Harms; Xavier Levecq - Web page: https://patentgrep.com/patent/US12216056B2 ## CPC - G01J 9/00: Measuring optical phase difference (devices or arrangements for controlling the phase of light beams G02F1/01); Determining degree of coherence; Measuring optical wavelength (spectrometry G01J3/00) - G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)} - G01J 3/4406: {Fluorescence spectrometry} - 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 21/365: {Control or image processing arrangements for digital or video microscopes (G02B21/361, G02B21/362 take 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} - H04N 23/56: provided with illuminating means - G01J 2009/002: {Wavefront phase distribution} - G01N 2201/06113: Coherent sources; lasers - G01J: MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY (light sources F21, H01J, H01K, H05B; investigating properties of materials by optical means G01N) - 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 - H04N: PICTORIAL COMMUNICATION, e.g. TELEVISION ## Abstract According to one aspect, the present description relates to a device for analysing a wavefront, configured to be connected to a fluorescence microscopy imaging system with optical sectioning, equipped with a microscope objective comprising a pupil in a pupil plane, the analysis device comprising a two-dimensional detector comprising a detection plane; a two-dimensional arrangement of microlenses, arranged in an analysis plane, each microlens being configured to form, on the detection plane, when the analysis device is connected to the microscopic imaging system, an image of an object situated in a focal plane of the microscope objective, with a given analysis field; an optical relay system configured to optically conjugate the analysis plane and the pupil plane; a field diaphragm positioned in a plane optically conjugated with the plane of detection, and configured to define said analysis field; a processing unit configured to determine, based on the set of images formed by the microlenses, a two-dimensional map of a characteristic parameter of the wavefront in said analysis plane. ## Claims ### Claim 1 A fluorescence microscopic imaging system with optical sectioning of a volumetric and fluorescent object comprising: an illumination path for an illumination of the object; an imaging path for imaging an optical section of the object, comprising a microscope objective lens with a pupil in a pupil plane and an imaging detector comprising an imaging detection plane, said optical section being superimposed on a focal plane of said microscope objective lens; an analysis path comprising said microscope objective lens and a wavefront analysis device configured to analyze a wavefront originating from the object; a beam splitter element for splitting the wavefront analysis device for analyzing optical defects from the imaging path; wherein the wavefront analysis device comprises: a two-dimensional detector comprising a detection plane conjugated with said focal plane of the microscope objective lens; a two-dimensional arrangement of a plurality of microlenses arranged in an analysis plane, each microlens of the plurality of microlenses being configured to form, on the detection plane, an image of said object, with an analysis field-of-view; an optical relay system configured to optically conjugate the analysis plane and the pupil plane; a field diaphragm positioned in a plane optically conjugated with the detection plane and configured to define said analysis field-of-view; a processing unit configured to determine, based on a set of images formed by the plurality of microlenses, a two-dimensional map of a characteristic parameter of a wavefront in said analysis plane, and wherein: the illumination path is configured for a first illumination of the object for fluorescence excitation for the imaging path, and a second illumination of the object for fluorescence excitation for the analysis path, the second illumination producing a structured illumination of the object according to a cross-shaped two-dimensional pattern. ### Claim 2 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein each microlens among the plurality of microlenses has a square pupil and the plurality of microlenses are joined together. ### Claim 3 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein each microlens among the plurality of microlenses comprises a pupil of a given shape, the field diaphragm having an identical shape to a shape of said pupil of each microlens. ### Claim 4 (depends on 3) The fluorescence microscopic imaging system as claimed in claim 3, further comprising one or more optical elements allowing optical conjugation to be carried out between the plane of the field diaphragm and the detection plane and wherein a plurality of dimensions of the field diaphragm are less than or equal to a plurality of dimensions of said pupil of each microlens, divided by the optical magnification (G) defined by the one or more optical elements. ### Claim 5 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein the field diaphragm has variable dimensions. ### Claim 6 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein the field diaphragm has a variable position in said plane optically conjugated with the detection plane. ### Claim 7 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein the field diaphragm is spatially structured with a given spatial frequency in one direction. ### Claim 8 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein the two-dimensional detector comprises a two-dimensional arrangement of elementary detectors and a diffraction spot of a microlens comprises, in one direction, between 0.2 and 2 elementary detectors. ### Claim 9 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein said beam splitter element is a dichroic element allowing light to be reflected in a first spectral band and light to be transmitted in a second spectral band, separate from the first spectral band. ### Claim 10 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, further comprising a wavefront correction device comprising a correction plane optically conjugated with the pupil plane, said correction plane being included in the analysis path, the wavefront correction device being configured to correct a wavefront originating from the object and analyzed by the wavefront analysis device. ### Claim 11 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein the illumination path is configured to form said optical section by transverse illumination of the object. ### Claim 12 (depends on 1) The fluorescence microscopic imaging system as claimed in claim 1, wherein the illumination path comprises: one or more laser sources for emitting ultrashort pulses, said microscope objective lens allowing each pulse to be focused at a focusing point of a focal plane of said microscope objective lens in order to form a multiphoton fluorescence emission; a scanning device configured to transversely scan said focusing point. ### Claim 13 (depends on 1) A method for microscopic imaging of an object by means of a fluorescence microscopic imaging system with optical sectioning according to claim 1, wherein the method comprises: illuminating the object along an optical section by means of the illumination path; imaging said optical section of the object in the detection plane of the imaging detector of the imaging path; and analyzing a wavefront originating from said optical section of the object by means of the analysis path comprising the microscope objective lens and the wavefront analysis device, wherein the illumination of the object comprises a first illumination for fluorescence excitation for the imaging path, and a second illumination for fluorescence excitation for the analysis path, the second illumination producing a structured illumination of the object according to a cross-shaped two-dimensional pattern.