Apparatus, Method, and System for the Automated Imaging and Evaluation of Embryos, Oocytes and Stem Cells

• US 20140106389A1
• Filed: 09/27/2011
• Published: 04/17/2014
• Est. Priority Date: 09/27/2010
• Status: Active Grant

• Current Assignee: Ares Trading SA (Merck & Co., Inc.)
• Original Assignee: Auxogyn Incorporated (Progyny Inc.)
• Inventors: Moussavi, Farshid, Loewke, Kevin, Young, Lisa, Menard, Edward, Shen, Shehua, Goldenstein, Lissa, Tucker, Eugene
• Granted Patent: US 9,482,659 B2
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: B01L 2200/026 Fluid interfacing between devices or objects, e.g. connectors, inlet details
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/143 Quality control, feedback systems
• CPC Class Codes: B01L 2300/024 Storing results with means integrated into the container
• CPC Class Codes: B01L 2300/0893 having a very large number of wells, microfabricated wells
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: C12M 21/06 for in vitro fertilization
• CPC Class Codes: C12M 41/14 Incubators; Climatic chambers per se B01L1/00
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12Q 3/00 Condition responsive control processes apparatus therefor C12M1/36; controlling or regulating in general G05
• CPC Class Codes: G01N 33/4833 of solid biological material, e.g. tissue samples, cell cultures tissue in vivo A61B5/00; cell suspensions G01N33/48735
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 35/00584 Control arrangements for automatic analysers
• CPC Class Codes: G02B 21/0088 Inverse microscopes
• CPC Class Codes: G02B 21/088 for both incident illumination and transillumination
• CPC Class Codes: G02B 21/125 affording both dark- and bright-field illumination
• CPC Class Codes: G02B 21/18 Arrangements with more than one light path, e.g. for comparing two specimens
• CPC Class Codes: G02B 21/365 Control or image processing arrangements for digital or video microscopes G02B21/361, G02B21/362 take precedence

Abstract

Apparatuses, methods, and systems for the automated imaging and evaluation of human embryos, oocytes, or pluripotent cells are described. An apparatus and method for automated dish detection and well occupancy determination are described. In addition, a multi-well culture dish and an illumination assembly for bimodal imaging are described. These inventions find use at least in identifying or in facilitating identification of embryos and oocytes in vitro that are most useful in treating infertility in humans.

Claims

1. An apparatus for automated imaging and evaluation of human embryos, oocytes, or pluripotent cells for use with an incubator, comprising:

at least one housing;

at least one time-lapse microscope placed inside the housing and having at least one light source and at least one imaging camera;

at least one loading platform extending outward from the housing, the loading platform for securing a multi-well culture dish holding a plurality of human embryos or pluripotent cells;

a computer for storing images from the at least one imaging camera and programmed for analyzing image sequences over time; and

at least one touch screen panel coupled to the computer and displaying a graphical user interface for controlling the at least one time-lapse microscope.

17. The apparatus of claim 1, wherein the computer is programmed to monitor the loading of the multi-well culture dish into the loading platform and adjust for misalignments of the multi-well culture dish into the loading platform.

2-16. -16. (canceled)

18-25. -25. (canceled)

26. A method for the automated imaging and evaluation of human embryos, oocytes, or pluripotent cells, comprising:

placing at least one human embryo or pluripotent cell in a multi-well culture dish;

loading the multi-well culture dish in a loading platform of an imaging system having at least one time-lapse microscope inside a housing;

if needed, adjusting the loading of the multi-well culture dish into the loading platform to verify a position and orientation of the multi-well culture dish;

acquiring time-lapse images of the multi-well culture dish;

displaying the images captured by the at least one time-lapse microscope in a graphical user interface accessible by a touch-screen panel; and

analyzing the time-lapse images of the multi-well culture dish to determine a development potential of the at least one human embryo or pluripotent cell.

28. The method of claim 26, further comprising determining occupancy of a plurality of micro-wells included in the multi-well culture dish.

29. The method of claim 26, further comprising determining parameters related to the development potential of the at least one human embryo or pluripotent cell.

30. The method of claim 29, wherein the parameters include a duration of first cytokinesis.

27. (canceled)

31-32. -32. (canceled)

33. A multi-well culture dish, comprising:

a ring disposed on a lower surface of the culture dish, the ring defining a cavity and having an upper surface, an outer lateral surface, and an inner lateral surface, the cavity having a cavity bottom;

a plurality of micro-wells defined by the cavity bottom, each micro-well configured to hold a human embryo or a pluripotent cell;

wherein;

the inner lateral surface of the ring is disposed between the outer lateral surface and the plurality of micro-wells, and extends from the upper surface of the ring to the cavity bottom; and

the inner lateral surface of the ring slopes toward the plurality of micro-wells such that a first width of the ring at the lower surface of the culture dish is greater than a second width of the ring at the upper surface of the ring.

35. The multi-well culture dish of claim 33, wherein a lower surface of at least one of the plurality of micro-wells is curved.

34. (canceled)

36-47. -47. (canceled)

48. An illumination assembly for bimodal imaging, comprising:

a first light source;

a condenser lens;

a darkfield aperture having a first surface configured to block light and having a second surface opposite to the first surface, the darkfield aperture defining at least one opening; and

a second light source attached to the second surface of the darkfield aperture;

wherein in a first mode of the illumination assembly, the first light source generates light that traverses the at least one opening in the darkfield aperture and the condenser lens prior to reaching a sample, and the second light source does not generate light; and

wherein in a second mode of the illumination assembly, the second light source generates light that reaches the sample without traversing the at least one opening in the darkfield aperture, and the first light source does not generate light.

49. The illumination assembly of claim 48, wherein:

the illumination assembly is configured in the first mode to perform time-lapse darkfield imaging of at least one of a human embryo, an oocyte, or a pluripotent cell; and

after completion of the time-lapse darkfield imaging, the illumination assembly is configured in the second mode to perform brightfield imaging of the at least one of a human embryo, an oocyte, or a pluripotent cell.

50-52. -52. (canceled)

53. An apparatus for automated dish detection and well occupancy determination, comprising:

a dish detection module configured to detect presence of a multi-well culture dish in an image detected by an imaging camera;

a well location determination module configured to determine a position of each of a plurality of micro-wells included in the multi-well culture dish;

a well occupancy determination module configured to determine occupied micro-wells included in the plurality of micro-wells based on the position of the each of the plurality of micro-wells; and

a display module configured to display at least the occupied micro-wells;

at least one of the dish detection module, the well location determination module, the well occupancy determination module, or the display module being implemented in at least one of a memory or a processing device.

57. The apparatus of claim 53, wherein the well location determination module is further configured to:

determine an initial position and an initial orientation of the each of the plurality of micro-wells in a first coordinate system;

determine the position of the each of the plurality of micro-wells in a second coordinate system based on the initial orientation of the each of the plurality of micro-wells, wherein the second coordinate system is rotated relative to the first coordinate system; and

determine a spatial extent of the each of the plurality of micro-wells in the second coordinate system based on the position of the each of the plurality of micro-wells in the second coordinate system.

54-56. -56. (canceled)

58-74. -74. (canceled)

75. An apparatus for automated imaging of human embryos, oocytes, or pluripotent cells, comprising:

a culture chamber configured to incubate a multi-well culture dish, the culture chamber having an upper surface including a first window and a lower surface including a second window;

a time-lapse microscope including a light source and an imaging camera configured to generate images of the multi-well culture dish inside the culture chamber based on light from the light source passing through the first window and the second window, wherein the culture chamber and the time-lapse microscope are integrated in a common housing; and

a touch-screen panel configured to display a graphical user interface for controlling the time-lapse microscope.

79. The apparatus of claim 75, further comprising an electronic watchdog circuit, wherein the electronic watchdog circuit is configured to:

measure a time duration that the light source has been on; and

shut off the light source if the time duration is longer than a threshold.

76-78. -78. (canceled)

80. (canceled)

81. A system for automated imaging and evaluation of human embryos, oocytes, or pluripotent cells for use with an incubator, comprising:

a plurality of imaging microscopes, each of the plurality of imaging microscopes being located inside a corresponding one of a plurality of housings and including at least one light source and at least one imaging camera, wherein each of the plurality of housings is located inside the incubator;

a loading platform extending outward from each of the plurality of housings, the loading platform for securing a multi-well culture dish holding a plurality of human embryos or pluripotent cells;

a controller electrically connected to each of the plurality of imaging microscopes, wherein the controller is located outside of the incubator and controls the at least one light source; and

a computer for storing images from the at least one imaging camera and programmed for analyzing image sequences over time, wherein the computer is electrically connected to each of the plurality of imaging microscopes via the controller.

89. The system of claim 81, wherein:

the at least one imaging camera is configured to generate images of a plurality of micro-wells included in the multi-well culture dish based on light from the at least one light source traversing the multi-well culture dish; and

the controller is configured to measure a time duration that the at least one light source has been on, and to shut off the at least one light source if the time duration is longer than a threshold.

82-88. -88. (canceled)

90-95. -95. (canceled)

96. A method for automated evaluation and display of human embryos, oocytes, or pluripotent cells, comprising:

collecting images of a plurality of multi-well culture dishes, each of the plurality of multi-well culture dishes including a plurality of micro-wells, at least one of the plurality of micro-wells containing at least one of a human embryo or a pluripotent cell;

analyzing the images of the plurality of multi-well culture dishes; and

concurrently displaying status information associated with each of the plurality of multi-well culture dishes.

97. The method of claim 96, wherein the status information includes:

patient information associated with each of the plurality of multi-well culture dishes;

a first status of collection of the images of each of the plurality of multi-well culture dishes; and

a second status of analysis of the images for each of the plurality of multi-well culture dishes.

98-105. -105. (canceled)

106. A system for automated imaging and evaluation of human embryos, oocytes, or pluripotent cells, comprising:

a plurality of time-lapse microscopes, each of the plurality of time-lapse microscopes being located inside a corresponding one of a plurality of housings and including at least one light source and at least one imaging camera, wherein each of the plurality of housings is located inside the incubator;

a loading platform extending outward from each of the plurality of housings, the loading platform for securing at least one multi-well culture dish holding a plurality of human embryos or pluripotent cells;

a computer electrically connected to the plurality of time-lapse microscopes; and

a server configured to communicate with the computer over a network, and configured to display a graphical user interface that provides status information and parameters determined based on analysis of images of a human embryo or pluripotent cell contained in the at least one multi-well culture dish, wherein the status information is associated with each of the plurality of time-lapse microscopes, and wherein at least one of the images is generated by each of the plurality of time-lapse microscopes.

107. The system of claim 106, wherein the status information includes:

patient information associated with the at least one multi-well culture dish;

a first status of collection of the images of the at least one multi-well culture dish; and

a second status of analysis of the images for the at least one multi-well culture dish.

108-111. -111. (canceled)