IMAGING AND EVALUATING EMBRYOS, OOCYTES, AND STEM CELLS
Methods, compositions and kits for determining the developmental potential of one or more embryos or pluripotent cells and/or the presence of chromosomal abnormalities in one or more embryos or pluripotent cells are provided. These methods, compositions and kits find use in identifying embryos and oocytes in vitro that are most useful in treating infertility in humans.
1 - 37 . (canceled)
38 . A method for assessing human embryo developmental potential in vitro in an automated system comprising the steps of:
a) capturing periodic sequential images of a human embryo over a 1-5 day period of human embryo development using one or more microscopes configured in an incubator that are operably linked to a computer comprising imaging software to capture the sequential images; and
b) using image analysis software configured on the computer to determine a cellular activity parameter from the sequential images of step, wherein the cellular activity parameter comprises the timing interval between the first and second mitosis of the human embryo and the timing interval between the second and third mitosis of the human embryo; and
c) using software configured on the computer to assess the developmental potential of the human embryo from the cellular activity parameter of step b.
39 . The method of claim 38 wherein the sequential images are captured every 1-30 minutes.
40 . The method of claim 38 wherein the sequential images are captured every 5 minutes.
41 . An automated system for assessing human embryo developmental potential in vitro comprising:
a) an incubator;
b) one or more microscopes configured in the incubator each comprising a camera capable of capturing images from the microscope;
c) a computer comprising:
i) imaging software capable of capturing sequential images from the one or more microscopes periodically over a 1-5 day period of human embryo development; and
ii) image analysis software capable of determining a cellular activity parameter from the sequential images of step e) i), and determining the developmental potential of the human embryo from the cellular activity parameter, wherein the cellular activity parameter comprises the timing interval between the first and second mitosis of the human embryo and the timing interval between the second and third mitosis of the human embryo.
42 . The automated system of claim 41 , further comprising image analysis software configured to compare the pixel values of sequential pairs of images to determine the amount of cellular activity for the most recent image of the pair by comparing pixel intensities of the most recent image to the pixel intensity of the previous image of the pair.
43 . The automated system of claim 41 wherein the cellular activity parameters are measured manually.
44 . The automated system of claim 41 , wherein the cellular activity parameters are measured automatically.
45 . The automated system of claim 41 , wherein the one or more microscopes employ darkfield illumination.
46 . The automated system of claim 41 wherein the images are captured every 1-30 minutes.
47 . The automated system of claim 41 , wherein the images are captured every 5 minutes.
48 . The automated system of claim 41 , comprising a single microscope
49 . The automated system of claim 41 , comprising an array of microscopes.
50 . The automated system of claim 41 , wherein the image analysis software is configured to determine a cellular activity parameter selected from the group consisting of cell size, thickness of the zona pellucida, degree of fragmentation, symmetry of daughter cells and duration of at least one cytokinesis.
51 . The automated system of claim 41 , wherein the image analysis software is configured to determine the time interval between the observation of a cleavage furrow and the resolution of cleavage furrow into two daughter cells.
52 . The automated system of claim 41 , wherein the so image analysis software is configured to determine the duration of a cell cycle event.
53 . The automated system of claim 41 , wherein the image analysis software is configured to assess embryos selected from the group consisting of one cell embryos, two cell embryos, three cell embryos, four cell embryos, 5 cell embryos and 6 cell embryos.
54 . The automated system of claim 41 , wherein the image analysis software comprises an algorithm for automated tracking of cell division.
55 . The automated system of claim 41 , wherein the image analysis software models embryos as a collection of ellipses with position, orientation and overlap index.
56 . The automated system of claim 41 , wherein the one or more microscopes track multiple embryos.
57 . The automated system of claim 41 wherein the timing interval between the first and second mitosis comprises the timing interval between the resolution of cytokinesis 1 and the onset of cytokinesis 2 or the resolution of cytokinesis 1 and the resolution of cytokinesis 2.
58 . The automated system of claim 41 wherein the timing interval between the second and third mitosis comprising the timing interval between the resolution of cytokinesis 2 and the onset of cytokinesis 3 or the resolution of cytokinesis 2 and the resolution of cytokinesis 3.
59 . The automated system of claim 41 wherein the image analysis software is further configured to measure the duration of cell cycle 1, cell cycle 2, cell cycle 3, and cell cycle 4.
60 . The automated system of claim 41 wherein good developmental potential is indicated by a time interval between the first and second mitosis of 8 to 15 hours.
61 . The automated system of claim 41 wherein good developmental potential is indicated by a time interval between the second and third mitosis of 0 to 5 hours.
62 . The automated system of claim 41 wherein good developmental potential is indicated by a time interval between the first and second mitosis of 8 to 15 hours and a time interval between the second and third mitosis of 0 to 5 hours.
63 . The automated system of claim 59 wherein good developmental potential is indicated by a duration of cell cycle 1, cell cycle 2, cell cycle 3, and cell cycle 4 that takes place within 54 hours and results in a 5 cell embryo.
64 . The automated system of claim 63 wherein good developmental potential is indicated by a duration of cell cycle 1, cell cycle 2, cell cycle 3, and cell cycle 4 that takes place within 54 hours and results in a 5 cell embryo and a time interval between the first and second mitosis of 8 to 15 hours.
65 . The automated system of claim 63 wherein good developmental potential is indicated by a duration of cell cycle 1, cell cycle 2, cell cycle 3, and cell cycle 4 that takes place within 54 hours and results in a 5 cell embryo and a time interval between the second and third mitosis of 0 to 5 hours.
66 . An automated system for assessing human embryo developmental potential in vitro comprising:
iii) an incubator;
iv) one or more microscopes configured in the incubator each comprising a camera capable of capturing images from the microscope;
v) a computer configured to assess embryo development parameters wherein the computer comprises:
i) imaging software capable of capturing sequential pairs of images from the one or more microscopes every 1-30 minutes over a 1-5 day period of human embryo development;
ii) image analysis software capable of determining a cellular activity parameter comprising the timing interval between the first and second mitosis of the human embryo and the timing interval between the second and third mitosis of the human embryo and further capable of determining the developmental potential of the human embryo from the cellular activity parameter, wherein good developmental potential is indicated by a time interval between the first and second mitosis of 8 to 15 hours and/or a time interval between the second and third mitosis of 0 to 5 hours.