# US 2019/0376012 A1 · An apparatus for the incubation of a biological material > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0376012 A1 - Document ID: US20190376012A1 - Status: Published application - Kind: A1 - Application number: 16/312,025 - Filed: 2017-06-30 - Published: 2019-12-12 - Earliest claimed: 2016-06-30 - Assignees: ESCO Medical Aps - Applicants: Esco Medical Aps - Inventors: Thomas William PEDERSEN - Web page: https://patentgrep.com/patent/US20190376012A1 ## CPC - C12M 29/26: {Conditioning fluids entering or exiting the reaction vessel} - G01N 21/6408: {with measurement of decay time, time resolved fluorescence} - C12M 37/02: {Filters} - C12M 41/36: {of biomass, e.g. colony counters or by turbidity measurements (electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404)} - C12M 41/48: {Automatic or computerized control (automatic analysis G01N35/00)} - C12M 21/06: {for in vitro fertilization} - C12M 25/06: {Plates; Walls; Drawers; Multilayer plates} - C12M 41/12: {of temperature (controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00)} - C12M 41/34: {of gas} - C12M 41/38: {of metabolites or enzymes in the cells} - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} - 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) ## Abstract The invention relates to an apparatus (200) for incubation of a viable biological material (2); said apparatus comprises: a housing (4) having an extension in a longitudinal direction X, in a transversal direction Y, and in a direction Z perpendicular to the longitudinal direction and the transversal direction; said housing comprising: two or more culture dish compartments (6), each being adapted to accommodate, one or more culture dishes (8) comprising a biological material (2); wherein said apparatus comprises an image capturing device (10); wherein said apparatus comprises a control unit (12) for controlling the operation thereof; wherein at least part of said image capturing device is being configured to be movable in relation to the two or more culture dish compartments (6), thereby allowing capture of images of one or more of said biological materials (2) accommodated in said one or more culture dishes (8); and wherein said apparatus comprises a FLIM unit (11) (fluorescent lifetime imaging microscope); wherein at least part of said FLIM unit (11) is being configured to be movable in relation to the two or more culture dish compartments (6), thereby allowing capture of FLIM spectra of one or more of said biological materials (2) accommodated in said one or more culture dishes (8). ## Claims ### Claim at position 1 1-55. (canceled) ### Claim 56 An apparatus (200) for incubation of a viable biological material (2); said apparatus comprises: a housing (4) having an extension in a longitudinal direction X, in a transversal direction Y, and in a direction Z perpendicular to the longitudinal direction and the transversal direction; said housing comprising: two or more culture dish compartments (6), each being adapted to accommodate, one or more culture dishes (8) comprising a biological material (2); wherein said apparatus comprises an image capturing device (10); wherein said apparatus comprises a control unit (12) for controlling the operation thereof; wherein at least part of said image capturing device is configured to be movable in relation to the two or more culture dish compartments (6), thereby allowing capture of images of one or more of said biological materials (2) accommodated in said one or more culture dishes (8); and wherein said image capturing device (10) is being a FLIM unit (fluorescent lifetime imaging microscope); wherein at least part of said FLIM unit (11) is being configured to be movable in relation to the two or more culture dish compartments (6), thereby allowing capture of FLIM spectra of one or more of said biological materials (2) accommodated in said one or more culture dishes (8): wherein said FLIM unit independently comprises: a laser source, such as a pulsed laser source, such as a diode laser or a multiphoton excitation laser; and optionally also one or more of the following elements: a single photon sensitive detector; a dichroic mirror (for separation of fluorescence signal from the excitation light); an objective (for focusing excitation light into sample and/or for collecting fluorescence signal); a control system for controlling said FLIM unit; wherein said laser source being coupled to optical means for conveying electromagnetic radiation in the form of one or more optical fibers, said optical means comprising a distal end being configured to be directed close to the biological material, for conveying by transmission, electromagnetic radiation. ### Claim 57 (depends on 56) An apparatus (200) according to claim 56, wherein one or more of said individual culture dish compartments (6), preferable all said culture dish compartments comprises its own individual lid (14), wherein each said lid is configured to be able to shift between an open configuration providing access to the corresponding culture dish compartment (6) and a closed configuration in which the corresponding culture dish compartment is being sealed off from the surroundings. ### Claim 58 (depends on 56, 57) An apparatus (200) according to any of claims 56-57 furthermore comprising temperature regulating means (16) for individual and independent regulation of the temperature in one or more of said individual culture dish compartments (6), preferable in each of said individual culture dish compartments. ### Claim 59 (depends on 56, 57, 58) An apparatus (200) according to any of claims 56-58 furthermore comprising gas composition regulating means (22) for individually regulating the gas composition, such as the concentration of oxygen, carbon dioxide and/or nitrogen in one or more of said individual culture dish compartments (6), preferable in each of said individual said culture dish compartments, and furthermore comprising one or more conduits (42) for conducting a gas from a gas mixing box (40) to one or more of said separate culture dish compartments (6). ### Claim 60 An apparatus (200) according to any of the preceding claims wherein in respect of one or more of said individual culture dish compartments (6), preferably in respect of each individual culture dish compartment, said individual culture dish compartment comprises a transparent shelf (24) for carrying a culture dish; and wherein said FLIM unit, or at least a light transmitting and a light receiving part of it, is/are arranged below said shelves and being adapted to be movable so as to enable capturing electromagnetic radiation, through said shelves (24), from said biological material (2) accommodated in any of said culture dish (8) being accommodated in any of said culture dish compartments (6). ### Claim 61 An apparatus (200) according to any of the preceding claims, wherein said FLIM unit independently comprises one or more of the following elements: a laser source in the form of a pulsed laser source, such as a diode laser or a multiphoton excitation laser; a dichroic mirror (for separation of fluorescence signal from the excitation light); an objective (for focusing excitation light into sample and/or for collecting fluorescence signal); a control system for controlling said FLIM unit. ### Claim 62 An apparatus (200) according to any of the preceding claims, wherein said FLIM unit is configured for auto-fluorescence of nicotine amide adenine (NADH) and/or for auto-fluorescence of flavine adenine dinucleotide (FAD) being involved in the metabolism of the biological material. ### Claim 63 An apparatus (200) according to any of the preceding claims wherein said apparatus being configured for operating in a time-correlated single photon counting (TCSPC) mode. ### Claim 64 An apparatus (200) according to any of the preceding claims wherein said apparatus being configured for operating in a FRET mode or in a FRAP mode or in a PLIM mode (Phosphorescence Lifetime Imaging Microscopy). ### Claim 65 An apparatus (200) according to any of the preceding claims wherein said apparatus during the fluorescence operations is being configured to operate in the time domain or in the frequency domain. ### Claim 66 (depends on 56, 57, 58, 59, 60, 61, 62, 63, 64, 65) A system (300) for incubation of a viable biological material; said system comprises: an apparatus (200) according to any of the claims 56-65, in combination with one or more culture dishes (8). ### Claim 67 An apparatus (200) according to any of the preceding claims wherein said apparatus is being configured to incubate two or more biological material in the form of oocytes or embryos; wherein said apparatus in respect of each specific of said two or more biological materials is being configured, based on images captured by said image capturing device (10), to identify a predetermined morphological state of said specific biological material; and 5 wherein said apparatus is being configured, in the event that a predetermined morphological state has been reached in respect of a specific biological material, to capture a FLIM spectrum of said specific biological material. ### Claim 68 (depends on 67) An apparatus (200) according to claim 67, wherein said apparatus is being configured, in respect of each specific of said two or more biological materials, to identify two or more 10 different and predetermined morphological states, based on said images captured by said image capturing device (10), of that specific biological material, and wherein in the event of each such different and predetermined morphological state has been reached in respect of a specific biological material, said apparatus is being configured to capture a FLIM spectrum of said specific biological material in respect of each such predetermined morphological state. ### Claim 69 (depends on 68) An apparatus (200) according to claim 68, wherein said apparatus in respect of each of said two or more biological materials is being configured for analyzing differences of the FLIM spectra corresponding to two or more different morphological states of the same biological material. ### Claim 70 (depends on 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66) Use of an apparatus (200) according to any of the claims 56-65 or of a system (300) according to claim 66 for incubation of a viable biological material (2). ### Claim 71 (depends on 67) Use according to claim 67 for empirically determining optimum protocols relating to physical and/or chemical conditions of a biological material (2) during the incubation thereof. ### Claim 72 (depends on 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66) A method for assessing optimum incubation conditions for a viable biological material, said method comprising the following steps: i) providing an apparatus (200) according to any of the claims 56-65 or a system (300) according to claim 66; ii) accommodating at least two culture dishes, each comprising one or more viable biological materials in separate culture dish compartments of said apparatus; iii) incubating said biological materials at incubation conditions, wherein the physical and/or chemical incubation conditions in respect of biological material being accommodated in one culture dish compartment differs by one or more parameters from the physical and/or chemical incubation conditions in respect of biological material being accommodated in another culture dish compartment; iv) during step iii), using the FLIM unit of said apparatus for capturing FLIM images; v) assessment of the quality of the biological material based on said FLIM images. ### Claim 73 (depends on 69) A method according to claim 69, wherein the most optimum physical and/or chemical incubation conditions is being determined, based on said FLIM images.