Animal cell confluence detection method and apparatus
First Claim
1. A process for detecting the degree of confluence of animal cells being cultured in a biological sample container, comprising:
- arranging a biological sample container in an object position of an imaging station;
illuminating the object position with an optical source from below at an oblique angle;
collecting an image of the biological sample container arranged in the object position such that the image is taken in a dark field configuration where light from the optical source, if not scattered, does not contribute to the image; and
processing the image to determine the degree of confluence of the animal cells in the biological sample container.
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Abstract
The invention provides an apparatus and process for detecting the degree of confluence of animal cells being cultured in a well plate. A well plate is arranged in an imaging station and illuminated with a ring of LEDs, or other optical source, from below at an oblique angle. An image of the well is captured with a CCD camera or other detector from above or below, such that the well image is taken in a dark field configuration where light from the optical source, if not scattered, does not contribute to the well image. By the simple solution of illuminating wells of a well plate from below at an oblique angle, it has been found that many animal cell types can be imaged with sufficient contrast to allow cell identification and consequent cell area computation using image processing techniques, thereby allowing confluence to be determined of animal cells being cultured in well plates. This avoids the need for more complex optical imaging techniques, such as phase contrast microscopy.
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Citations
36 Claims
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1. A process for detecting the degree of confluence of animal cells being cultured in a biological sample container, comprising:
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arranging a biological sample container in an object position of an imaging station;
illuminating the object position with an optical source from below at an oblique angle;
collecting an image of the biological sample container arranged in the object position such that the image is taken in a dark field configuration where light from the optical source, if not scattered, does not contribute to the image; and
processing the image to determine the degree of confluence of the animal cells in the biological sample container. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. An apparatus for detecting the degree of confluence of animal cells being cultured in a biological sample container, comprising:
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an imaging station where a biological sample container can be arranged in an object position;
an optical source arranged to illuminate the object position from below at an oblique angle;
a detector arranged to collect an image of the biological sample container arranged in the object position such that the image is taken in a dark field configuration where light from the optical source, if not scattered, does not contribute to the image; and
an image processing unit for processing images to determine the degree of confluence of animal cells culturing in the biological sample container. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 35, 36)
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25. Use of a robot equipped with an animal cell picking head comprising at least one hollow pin for aspirating animal cells and replating them by moving them from a target well plate to a destination biological sample container under the control of a head positioning system, the use comprising:
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providing a well plate in which animal cells are being cultured;
arranging the well plate at an imaging station and detecting the degree of confluence in each well by repeatedly;
(i) illuminating a selected well from below at an oblique angle;
(ii) acquiring an image of the illuminated well; and
(iii) processing the image of the well to determine the degree of confluence; and
dependent on the degree of confluence, either replating the animal cells out of the well plate, or leaving them to continue to culture. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34)
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Specification