JP2005218379A5 - - Google Patents

Description

  The cultured cell state measurement method of the present invention that achieves the above object includes an illumination optical system, an observation optical system that includes a unit that converts an observation image into a contrast image by interference, and a sign of a phase difference when the observation image interferes The image having the imaging element that captures the opposite contrast image is captured by the microscope having two contrast images having opposite phase difference signs, and the difference calculation processing is performed by calculating the difference between the two captured images. The number of cells within the observation range, the occupied area of the cells, the distribution of cells, the density of cells, the density of cells, the morphology of the cells, the distribution of cell nuclei, and the distribution of scattering of cells from the formed and obtained difference calculation image It is a method characterized by measuring one of them.

  In this case, from the intensity distribution of the difference-calculated image, the portion where the intensity value is near zero is extracted to make the portion where the cell does not exist, and the other portion exists as the portion where the cell exists. It is desirable to separate the part from the non-existing part.

  Further, from the intensity distribution of the difference-processed image, a portion where the intensity value is near zero is extracted, the intensity value is assigned to one value, and another portion is assigned another intensity value to the value. You may make it binarize.

  Another culture cell state measurement method of the present invention includes an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and a sign of a phase difference at the time of interference of the observation image is opposite. A microscope having an image sensor that captures a contrast image captures an image composed of two contrast images with opposite phase difference signs, and forms two images obtained by performing a difference operation and a sum operation of the two captured images, respectively. Then, the difference-calculated image is divided by the sum-calculated image, and the cell thickness distribution and cell phase distribution within the observation range are measured.

  The cultured cell state measurement apparatus of the present invention includes an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image due to interference, and a contrast image in which the sign of the phase difference at the time of interference of the observation image is opposite. A microscope having an image pickup device that captures the image, and an image composed of two contrast images having opposite signs of the phase difference are captured by the microscope, and a difference calculation process is performed by calculating a difference between the two captured images. From the obtained difference-calculated image, at least one of the number of cells in the observation range, the cell occupation area, the cell distribution, the cell density, the cell morphology, the cell nucleus distribution, and the cell scattering distribution And a processing unit for measuring one.

  In this case, from the intensity distribution of the difference-calculated image, the portion where the intensity value is near zero is extracted to make the portion where the cell does not exist, and the other portion exists as the portion where the cell exists. It is desirable to have a separation processing unit that separates a portion from a portion that does not exist.

  Further, from the intensity distribution of the difference-processed image, a portion where the intensity value is near zero is extracted, the intensity value is assigned to one value, and another portion is assigned another intensity value to the value. A binarization processing unit for binarization may be provided.

  Another cultured cell state measuring apparatus of the present invention is an illumination optical system, an observation optical system including a unit for converting an observation image into a contrast image by interference, and a sign of a phase difference at the time of interference of the observation image. A microscope having an image pickup device that captures the contrast image, and an arithmetic unit that captures an image composed of two contrast images with opposite signs of phase difference by the microscope, and performs a difference operation and a sum operation of the two captured images. And a thickness measurement unit that divides the difference-calculated image by the sum-calculated image and measures the cell thickness distribution and the cell phase distribution within the observation range.

Claims (16)

A microscope having an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and an image pickup device that captures a contrast image having a phase sign opposite to that of the observation image when the interference is observed. An image composed of two contrast images having opposite phase signs is captured, the difference between the two captured images is calculated to form an image, and an image is formed from the obtained difference calculated image. The state of a cultured cell characterized by measuring at least one of the number of cells in the cell, the area occupied by the cell, the cell distribution, the cell density, the cell morphology, the cell nucleus distribution, and the cell scattering distribution Measurement method. From the intensity distribution of the difference-calculated image, a portion where the intensity value is near zero is extracted as a portion where cells do not exist, and the other portion as a portion where cells exist and a portion where cells exist does not exist The method for measuring a state of a cultured cell according to claim 1, wherein the portions are separated. From the intensity distribution of the difference-calculated image, a portion whose intensity value is near zero is extracted and assigned an intensity value to one value, and another portion is assigned another intensity value as a binary value. The method for measuring a state of a cultured cell according to claim 1, wherein: A microscope having an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and an image pickup device that captures a contrast image having a phase sign opposite to that of the observation image when the interference is observed. An image obtained by capturing an image composed of two contrast images having opposite phase signs, forming two images obtained by performing a difference operation and a sum operation of the two captured images, and performing a sum operation process on the difference-calculated image. And measuring the cell thickness distribution and the cell phase distribution within the observation range. A microscope having an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and an image pickup device that captures a contrast image having a phase sign opposite to that of the observation image when the interference is observed. An image composed of two contrast images having opposite phase signs is captured, the difference between the two captured images is calculated to form an image, and an image is formed from the obtained difference calculated image. A processing unit for measuring at least one of the number of cells in the cell, the area occupied by the cells, the cell distribution, the cell density , the cell morphology, the cell nucleus distribution, and the cell scattering distribution. A state measurement device for cultured cells. From the intensity distribution of the difference-calculated image, a portion where the intensity value is near zero is extracted as a portion where cells do not exist, and the other portion as a portion where cells exist and a portion where cells exist does not exist 6. The culture cell state measuring apparatus according to claim 5, further comprising a separation unit for separating the portion. From the intensity distribution of the difference-calculated image, a portion whose intensity value is near zero is extracted and assigned an intensity value to one value, and another portion is assigned another intensity value as a binary value. 6. The apparatus for measuring a state of cultured cells according to claim 5, further comprising a binarization unit for converting the information into a cell. A microscope having an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and an image pickup device that captures a contrast image having a phase sign opposite to that of the observation image when the interference is observed. An image obtained by capturing an image composed of two contrast images having opposite phase signs, forming two images obtained by performing a difference operation and a sum operation of the two captured images, and performing a sum operation process on the difference-calculated image. And a unit for measuring the cell thickness distribution and the cell phase distribution within the observation range. An opening is disposed at a substantially pupil position of the illumination optical system, and an observation optical system including a unit that converts the observation image into a contrast image by interference is illuminated by light that has passed through the opening of the illumination optical system. And an observation optical system configured to capture the image of the observation object by the image sensor at each of the positions shifted from the in-focus position to the near point side and the far point side, and The culture cell state measuring apparatus according to any one of claims 14 to 16 . A microscope having an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and an image pickup device that captures a contrast image having a phase sign opposite to that of the observation image when the interference is observed. By capturing an image composed of two contrast images having opposite phase difference signs, performing a difference operation between the two captured images, and performing a difference operation process to form an image, uneven illumination light within the observation range, The method for measuring a state of a cultured cell according to claim 1, wherein the influence of a foreign substance other than the cell to be removed is removed. A microscope having an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and an image pickup device that captures a contrast image having a phase sign opposite to that of the observation image when the interference is observed. An image obtained by capturing an image composed of two contrast images having opposite phase signs, forming two images obtained by performing a difference operation and a sum operation of the two captured images, and performing a sum operation process on the difference-calculated image. And measuring at least one of the height of the cell nucleus within the observation range, the area of the cell nucleus, and the flatness of the cell nucleus. 12. The threshold value is set to an image obtained by dividing the difference calculation image by the sum calculation image, and the image information is separated into an area within the threshold value and an area exceeding the threshold value. Of measuring the state of cultured cells. An opening is disposed at a substantially pupil position of the illumination optical system, and an observation optical system including a unit that converts the observation image into a contrast image by interference is illuminated by light that has passed through the opening of the illumination optical system. The difference image is formed from the two images captured by the image sensor at each position shifted from the in-focus position to the near point side and the far point side of the image of the observation object. And the state measuring method of the cultured cell of any one of Claims 10-12. A microscope having an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and an image pickup device that captures a contrast image having a phase sign opposite to that of the observation image when the interference is observed. By capturing an image consisting of two contrast images with opposite signs of phase difference, performing a difference operation between the two captured images, and performing a difference operation process to form an image, the illumination light unevenness within the range is targeted. 6. The cultured cell state measuring apparatus according to claim 5, which has a function of removing the influence of foreign substances other than cells. A microscope having an illumination optical system, an observation optical system including a unit that converts an observation image into a contrast image by interference, and an image pickup device that captures a contrast image having a phase sign opposite to that of the observation image when the interference is observed. An image obtained by capturing an image composed of two contrast images having opposite phase signs, forming two images obtained by performing a difference operation and a sum operation of the two captured images, and performing a sum operation process on the difference-calculated image. And a unit for measuring at least one of the height of the cell nucleus within the observation range, the area of the cell nucleus, and the flatness of the cell nucleus. 16. The image processing apparatus according to claim 8, further comprising a function of setting a threshold value to an image obtained by dividing the difference-calculated image by the sum-calculated image and separating the image information into an area within the threshold value and an area exceeding the threshold value. The state measurement apparatus of the cultured cell as described.