Culture-medium-monitoring apparatus

Patent metadata

Field	Value
Publication number	US-11299701-B2
Application number	US-202016802665-A
Country	US
Kind code	B2
Filing date	Feb 27, 2020
Priority date	Mar 19, 2019
Publication date	Apr 12, 2022
Grant date	Apr 12, 2022

Abstract

Provided is a culture-medium-monitoring apparatus including: an optical measurement unit that includes an illumination light source and a collecting lens that radiate an illumination light onto a culturing liquid, a retroreflective member that has an array in which micro-reflective elements are arrayed, that is disposed so as to sandwich the vessel between the retroreflective member, and the illuminating light source and the collecting lens, and that reflects the illumination light passed through the culturing liquid in the vessel, and a light detector that detects an intensity of the illumination light passed through the culturing liquid in the vessel after being reflected by the retroreflective member; and a control portion that causes the intensity of the illumination light to be repeatedly detected at a prescribed timing, and that determines a state of the culturing liquid on the basis of a change over time in the intensity of the illumination light.

First claim

The invention claimed is:

1. A culture-medium-monitoring apparatus comprising:
an optical measurement unit that includes:
an illuminating portion that is configured to radiate an illumination light beam onto a culture medium in a closed-bottom cylindrical vessel as a bioreactor;
a stirrer disposed in the vessel and configured to rotate around a center axis of the vessel to stir the culture medium in the vessel;
a retroreflective member that has an array in which a plurality of micro-reflective elements are arrayed, that is disposed so as to sandwich the vessel between the illuminating portion and the retroreflective member, and that is configured to reflect the illumination light beam that has passed through the culture medium in the vessel; and
a light-detecting portion that is configured to detect an intensity of the illumination light beam that has passed through the culture medium in the vessel after being reflected by the retroreflective member; and

a control portion that is configured to cause the intensity of the illumination light beam to be repeatedly detected at a prescribed timing by controlling the optical measurement unit, and to determine a state of the culture medium based on a change over time in the intensity of the illumination light beam,
wherein:
the illuminating portion is configured to enter the illumination light beam into the vessel along a direction intersecting a height direction of the vessel, and
the illumination portion, the retroreflective member, and the stirrer are disposed at positions at which the illumination light beam traveling inside the vessel does not interfere with the stirrer.

2. The culture-medium-monitoring apparatus according to claim 1 , further comprising:
a notifying portion that is configured to issue a notification about information to a user,
wherein the control portion issues, via the notifying portion, a notification about a timing for replacing the culture medium to the user.

3. The culture-medium-monitoring apparatus according to claim 1 , further comprising:
a culture-medium-supplying portion that is configured to supply the culture medium to the vessel; and
a culture-medium-discharging portion that is configured to discharge the culture medium from the vessel,
wherein, in a case in which the control portion determines that the timing for replacing the culture medium has arrived based on the change over time in the intensity of the illumination light beam detected by the light-detecting portion, the control portion causes the culture-medium-discharging portion to discharge a portion of the culture medium from the vessel, and causes the culture-medium-supplying portion to supply a new culture medium to the vessel.

4. The culture-medium-monitoring apparatus according to claim 1 , wherein the control portion causes a speed at which the culture medium is stirred by the stirrer to be reduced to move suspended cells in the culture medium below an optical path of the optical measurement unit by means of gravity when detecting the intensity of the illumination light beam by means of the light-detecting portion.

5. The culture-medium-monitoring apparatus according to claim 1 , wherein the optical measurement unit comprises a detection optical system that is configured to cause an image of cells suspended in the culture medium irradiated with the illumination light beam to be formed on the light-detecting portion.

6. The culture-medium-monitoring apparatus according to claim 5 , wherein the optical measurement unit comprises a phase contrast optical system that is configured to generate a phase contrast image of the cells.

7. The culture-medium-monitoring apparatus according to claim 5 , wherein the control portion repeatedly performs detection of the intensity of the illumination light beam and acquisition of the image of the cells by means of the light-detecting portion, causes the speed at which the culture medium is stirred by the stirrer to be reduced to move suspended cells in the culture medium below an optical path of the optical measurement unit by means of gravity when detecting the intensity of the illumination light beam, and causes the culture medium to be stirred without reducing the speed at which the culture medium is stirred by the stirrer to make the cells suspended in the culture medium when acquiring the image of the cells.

8. The culture-medium-monitoring apparatus according to claim 1 , wherein the illuminating portion comprises a monochromatic light source that is configured to emit the illumination light beam at a single wavelength.

9. The culture-medium-monitoring apparatus according to claim 1 , wherein the illuminating portion comprises a light source that is configured to emit, as the illumination light beam, a plurality of monochromatic light beams at different wavelengths, and
wherein the control portion determines the state of the culture medium based on changes over time in intensities of the light beams at the respective wavelengths that have passed through the culture medium and that are detected by the light-detecting portion.

10. The culture-medium-monitoring apparatus according to claim 1 , wherein:
the illuminating portion radiates the illumination light beam onto a specific region in which the cells and the culture medium are present in the vessel,
the light-detecting portion includes an image-acquisition portion that is configured to acquire an image of the specific region by capturing an image of a light beam, which is the illumination light beam that has been reflected by the retroreflective member and that has passed through the specific region in the vessel,
the control portion includes an image-analyzing portion that is configured to divide the image of the specific region acquired by the image-acquisition portion into pixels containing the cells and background pixels, and to calculate a representative pixel value that represents the background pixels, and
the representative pixel value calculated by the image-analyzing portion corresponds to the intensity of the illumination light beam.

11. The culture-medium-monitoring apparatus according to claim 1 , wherein the stirrer comprises a stirring rod and a plurality of stirring blades provided on the stirring rod, and the stirrer stirs the culture medium by rotating the stirring rod.

Assignees

• Olympus Corp

Inventors

• Sasaki Hiroshi

Classifications

• C12M41/00 Primary

  Means for regulation, monitoring, measurement or control, e.g. flow regulation (controlling or regulating chemical, physical or physicochemical processes B01J19/0006; heating or cooling apparatus for laboratory use B01L7/00; electro optical investigation of individual particles, flow cytometers G01N15/14; automatic analysis G01N35/00; controlling or regulating in general G06N) · CPC title

• H04N23/56

  provided with illuminating means · CPC title

• C12M41/06 Primary

  of illumination · CPC title

• G06T7/194

  involving foreground-background segmentation · CPC title

• G06T2207/10024

  Color image · CPC title