Apparatus, method, and system for the automated imaging and evaluation of embryos, oocytes and stem cells

• US 9,482,659 B2
• Filed: 09/27/2011
• Issued: 11/01/2016
• Est. Priority Date: 09/27/2010
• Status: Active Grant

• Current Assignee: Ares Trading SA (Merck & Co., Inc.)
• Original Assignee: Progyny Inc.
• Inventors: Young, Lisa, Menard, Edward, Tucker, Eugene, Loewke, Kevin, Shen, Shehua, Goldenstein, Lissa, Moussavi, Farshid
• Primary Examiner(s): Bowers, Nathan
• Assistant Examiner(s): EDWARDS, LYDIA E
• Application Number:
  US13/823,704
• Publication Number:
  US 20140106389A1
• Time in Patent Office: 1,862 Days
• US Class Current: 1/1
• 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. A method for automated imaging and evaluation of human embryos, oocytes, or pluripotent cells with an imaging system, the method comprising:

receiving a multi-well culture dish comprising a plurality of micro-wells on a stage of the imaging system, wherein the plurality of micro-wells contain a plurality of human embryos or pluripotent cells, wherein the imaging system comprises-at least one time-lapse microscope inside a housing;

initializing the imaging system, wherein initializing the imaging system comprises;

generating an initial image of the multi-well culture dish with the imaging system;

detecting the presence of the multi-well culture dish on the stage of the imaging system;

generating an estimate of a position and an orientation of the plurality of micro-wells, wherein generating the estimate of the position and the orientation of the plurality of micro-wells comprises;

generating an initial estimate of micro-well position and orientation;

generating a refined estimate of micro-well position and orientation; and

determining the position and orientation of the micro-wells in a rotated coordinate system;

providing an alignment indication, wherein the alignment indication comprises a first indication if the multi-well culture dish is properly aligned and a second indication if the multi-well culture dish is improperly aligned;

automatically illuminating the multi-well culture dish with an illumination source of the at least one time-lapse microscope when the multi-well culture dish is properly aligned; and

automatically focusing the at least one time-lapse microscope configured to capture time-lapse images of the multi-well culture dish when the multi-well culture dish is properly aligned;

acquiring time-lapse images of the multi-well culture dish with a camera of the at least one time-lapse microscope; and

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

2. The method of claim 1, wherein initializing the imaging system further comprises determining occupancy of a plurality of micro-wells in the multi-well culture dish.

3. The method of claim 1, further comprising determining parameters related to the development potential of the at least one human embryo or pluripotent cell;

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

4. The method of claim 3, wherein the parameters include a duration of first cytokinesis.

5. The method of claim 1, wherein the alignment indication comprises a visual feedback comprising a color coded LED.

6. The method of claim 1, wherein generating the initial estimate of micro-well position and orientation comprises:

retrieving a template, wherein the template represents at least a portion of the plurality of micro-wells included in the multi-well culture dish;

rotating the template through a series of predetermined positions;

generating a normalized cross-correlation between the initial image and the template in each of the series of predetermined positions;

identifying one of the series of predetermined positions of the template with the highest normalized cross-correlation score; and

identifying the initial estimate of the micro-well position and orientation as the position and orientation of the template in the one of the series of predetermined positions of the template with the highest normalized cross-correlation score.

7. The method of claim 1, wherein generating the refined estimate of micro-well position and orientation comprises:

selecting a first set of features from the initial image, wherein the first set of features from the initial image correspond to a set of features of the multi-well culture dish;

generating vectors between adjacent pairs of features of the first set of features from the initial image; and

combining the vectors to determine the orientation of the multi-well culture dish.

8. The method of claim 1, wherein providing an alignment indication comprises determining an initial alignment of the multi-well culture dish by detecting the relative position of an indexing feature of the multi-well culture dish with respect to a feature of the stage of the imaging system.

9. A method for automated imaging and evaluation of human embryos, oocytes, or pluripotent cells with an imaging system, the method comprising:

receiving a multi-well culture dish comprising a plurality of micro-wells on a stage of the imaging system, wherein the plurality of micro-wells contain a plurality of human embryos or pluripotent cells, wherein the imaging system comprises-at least one time-lapse microscope inside a housing;

initializing the imaging system, wherein initializing the imaging system comprises;

generating an initial image of the multi-well culture dish with the imaging system;

detecting the presence of the multi-well culture dish on the stage of the imaging system;

determining occupancy of a plurality of micro-wells in the multi-well culture dish, wherein determining occupancy of a plurality of micro-wells included in the multi-well culture dish comprises;

selecting at least one of the plurality of micro-wells;

identifying pixels located in the selected at least one of the plurality of micro-wells;

determining the intensity of the identified pixels;

comparing the intensity of the identified pixels to a brightness threshold;

determining a percent of pixels in the at least one of the plurality of micro-wells that exceed the brightness threshold; and

identifying the at least one of the plurality of micro-wells as occupied if the percent of pixels is greater than a threshold value;

providing an alignment indication, wherein the alignment indication comprises a first indication if the multi-well culture dish is properly aligned and a second indication if the multi-well culture dish is improperly aligned;

automatically illuminating the multi-well culture dish with an illumination source of the at least one time-lapse microscope when the multi-well culture dish is properly aligned; and

automatically focusing the at least one time-lapse microscope configured to capture time-lapse images of the multi-well culture dish when the multi-well culture dish is properly aligned;

acquiring time-lapse images of the multi-well culture dish with a camera of the at least one time-lapse microscope; and

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