Cell culture device and image analysis device

Abstract

Provided is a method for non-invasively and quantitatively determining multilayerization and differentiation when culturing a cell sheet. Provided is a method for determining a cell state by imaging a cell sheet by using an optical instrument characterized by having a high resolution, and then analyzing the inner structure thereof.

1 . An image analysis device which non-invasively analyzes an image optically acquired from a cell sheet in which cells are stratified on a culture surface, comprising:
 an acquisition part which acquires a plurality of cross-sectional images taken at different distances from the culture surface in the stratification direction,   a measuring part which measures the number of cells contained in each of the plurality of cross-sectional images, and an analyzing part which analyzes the distribution of the number of cells at least in the stratification direction based on the number of cells.   
     
     
         2 . The image analysis device according to  claim 1 , wherein the analyzing part determines the position of each of the plurality of layers which constitute a stratified cell sheet from the distribution of the number of cells in the stratification direction. 
     
     
         3 . The image analysis device according to  claim 1 , wherein the analyzing part calculates the distance between cells contained in the plurality of cross-sectional images, and determines the position of each of the plurality of layers which constitute the stratified cell sheet based on the distance between cells. 
     
     
         4 . The image analysis device according to  claim 2 , wherein the size of cells contained in at least one layer among the plurality of layers is determined, and when the size of the cells is greater than a predetermined value, it is determined that the cells contained in the at least one layer are differentiated cells. 
     
     
         5 . The image analysis device according to  claim 1 , wherein a light source, a condensing optical system, and a detection optical system are provided, the condensing optical system has a light branching means which branches light from the light source into a first signal light and a first reference light, and an objective lens which condenses the first signal light on the cell sheet and irradiates the cells,
 and the detection optical system has an interference optical system which multiplexes the first signal light and the first reference light and generates three or more interference lights having different phase relations, and an optical detector which detects the three or more interference lights and outputs a plurality of detection signals as electrical signals.   
     
     
         6 . The image analysis device according to  claim 5 , wherein the objective lens has a numerical aperture of 0.4 or more. 
     
     
         7 . The image analysis device according to  claim 5 , wherein the number of interference lights generated in the interference optical system is four, the interference phases of the first signal light and the first reference light are different only by the integral multiples of approximately 90 degrees, and
 a pair of interference lights in which the interference phases of the reference lights of the first signal light and the first interference light are different by approximately 180 degrees from each other is detected by a current differential type optical detector.   
     
     
         8 . The image analysis device according to  claim 1 , wherein the analyzing part calculates the density of cells based on the number of cells measured, generates a graph image in which a position in the stratification direction is defined on one axis and the density of cells in that position is defined on the other axis, and
 displays the graph image on a display device.   
     
     
         9 . The image analysis device according to  claim 1 , wherein the analyzing part
 measures the distances between cells or the sizes of the cells contained in each of the plurality of images from the information of the number of cells contained in each of the plurality of images, generates an image which is a graph where the distances between cells in the stratification direction or the distribution of the sizes of the cells are represented by defining the distance between cells or the sizes of the cells on one axis and the number of cells having those distances between cells or the sizes of the cells on the other axis, and   displays the graph image on a display device.   
     
     
         10 . The image analysis device according to  claim 1 , wherein the analyzing part generates analytical data regarding the distribution of the number of cells in the stratification direction from the information of the number of cells contained in each of the plurality of images, and performs at least one of:
 displaying the analytical data, generating an alarm based on the analytical data, and outputting a signal based on analytical data to the cell culture device or another external device.   
     
     
         11 . The image analysis device according to  claim 1 , wherein the analyzing part measures the distances between cells or the sizes of the cells contained in each of the plurality of images from the information of the number of cells contained in each of the plurality of images, generates analytical data regarding the distances between cells of cells or the distribution of the sizes of cells contained in the cell sheet, and
 performs at least one of: displaying the analytical data, generating an alarm based on the analytical data, and outputting a signal based on the analytical data to the cell culture device or to another external device.   
     
     
         12 . The image analysis device according to  claim 3 , wherein the size of cells contained in at least one layer among the plurality of layers is determined, and when the size of the cells is greater than a predetermined value, it is determined that the cells contained in the at least one layer are differentiated cells.