JPWO2019135267A1 - Cell image processing device, cell image processing method and cell image processing program - Google Patents

Abstract

The cell image processing apparatus (1) inputs a plurality of images acquired by photographing cells at time intervals, and processes and evaluates any two of the input images. A proliferation parameter calculation unit (4) that calculates a proliferation parameter representing a change in the amount of cells per unit time in a predetermined counting area of an image, and an inflow or outflow of cells per unit time in the counting area of each evaluation image. A migration parameter calculation unit (6) that calculates a migration parameter representing an amount, an area size calculation unit (8) that calculates the size of a counting area based on past growth parameters and past migration parameters, and an area size calculation. A proliferation rate calculation unit (7) for calculating a cell proliferation rate based on a growth parameter and a migration parameter calculated for a counting region having a size calculated by the unit (8) is provided.

Description

The present invention relates to a culture image processing apparatus, a cell image processing method, and a cell image processing program.

In order to quantitatively observe changes in cell proliferation over time, there is known a technique for analyzing an image of cells to obtain parameters representing changes in the state of cells (see, for example, Patent Document 1). Specifically, the timing of passage is determined based on the confluence rate, which is the ratio of the area occupied by cells in the image.

Further, in order to observe the relationship between cell aggregation and cell division such as colony formation, a technique is known for obtaining a statistic of the positional relationship between a pixel of interest and a peripheral pixel as a parameter representing cell aggregation (for example). , Patent Document 2).

WO2016 / 098271 JP-A-2009-229274

In order to control the culture of cells accurately, it is necessary to accurately detect whether or not the cells are proliferating. However, there is an inconvenience that cell proliferation cannot be detected accurately only by acquiring parameters such as the confluence rate as in Patent Document 1 and cell aggregation around the cells of interest as in Patent Document 2.

The present invention has been made in view of the above circumstances, and is a cell image processing apparatus, a cell image processing method, and a cell image capable of accurately detecting cell proliferation and accurately managing cell culture. The purpose is to provide a processing program.

In one aspect of the present invention, a plurality of images acquired by photographing cells at time intervals are input, and any two of the input images are processed for evaluation. A growth parameter calculation unit that calculates a growth parameter representing a change in the amount of the cells per unit time in a predetermined counting area of an image, and an inflow amount of the cells per unit time of the counting area of each evaluation image or A migration parameter calculation unit that calculates a migration parameter representing the outflow amount, a region size calculation unit that calculates the size of the counting area based on the past proliferation parameter and the past migration parameter, and the region size calculation unit. It is a cell image processing apparatus including a proliferation rate calculation unit which calculates a cell proliferation rate based on the proliferation parameter and the migration parameter calculated for the counting area of the size calculated by.

According to this aspect, when a plurality of images acquired by taking pictures at time intervals are input, any two of the input images are processed by the proliferation parameter calculation unit. The growth parameters in the counting area are calculated. In addition, the evaluation image is processed by the migration parameter calculation unit to calculate the migration parameter in the counting area.

Then, the region size calculation unit calculates the size of the counting region based on the past proliferation parameters and the past migration parameters, and proliferates based on the proliferation parameters and the migration parameters calculated for the counting region of the calculated size. The cell proliferation rate is calculated by processing by the rate calculation unit.
The cell proliferation rate is determined by taking into account not only the proliferation parameter indicating the change in cell mass in the two images separated by time but also the migration parameter indicating cell influx and cell outflow. The degree of cell proliferation can be detected more accurately, and cell culture can be managed accurately.

In this case, the proliferation parameter is proportional to the area of the counting region, and the migration parameter is proportional to the peripheral length of the counting region. Therefore, the smaller the counting region, the greater the influence of the migration parameter on the calculation of the cell proliferation rate, resulting in an error. Becomes larger. Therefore, the cell proliferation rate can be calculated accurately by calculating the cell proliferation rate based on the proliferation parameter and the migration parameter calculated for the counting region set to an appropriate size.

Here, the evaluation image and the growth measurement image may use the same image or different images. That is, the time interval between the two evaluation images and the time interval between the two growth measurement images may be the same or different. For example, as the evaluation image, all two adjacent images may be used, or two images selected periodically or aperiodically may be used.

In the above aspect, the growth parameter calculation unit may calculate the number of cells increased in the counting region per unit time as the growth parameter.
By doing so, it is easy to process the two evaluation images acquired at time intervals, count the number of cells existing in each counting area, and divide the difference by the time interval. Proliferation parameters can be calculated.

Further, in the above aspect, the migration parameter calculation unit may calculate the difference between the number of cells flowing in from the outer edge of the counting region and the number of cells flowing out from the outer edge of the counting region per unit time as the migration parameter.
By doing so, the two evaluation images acquired at time intervals are processed to count the number of cells that have flowed in and out of the outer edge of each counting area, and the difference is timed. The migration parameter can be easily calculated by dividing by the interval.

Further, in the above aspect, the region size calculation unit calculates the lower limit of the size of the counting region by comparing the ratio of the past migration parameters to the past proliferation parameters with a predetermined margin of error. May be good.
By doing so, the cell growth rate calculation unit can calculate the cell growth rate with high accuracy equal to or less than the margin of error.

Further, in the above aspect, the counting area may be rectangular.
By doing so, the proliferation parameter, the migration parameter and the cell proliferation rate can be easily calculated.

In addition, another aspect of the present invention includes a processor in which a plurality of images acquired by photographing cells at time intervals are input, and the processor is a unit time within a predetermined counting area in the past. The size of the counting region is calculated and calculated based on the proliferation parameter representing the change in the amount of the cells per hit and the migration parameter representing the inflow or outflow of the cells per unit time of the counting region in the past. A cell that processes any two of the input images using the counting region of the calculated size to calculate the proliferation parameter and the migration parameter in the counting region of the image. It is an image processing device.

Further, in another aspect of the present invention, a plurality of images acquired by photographing cells at time intervals are input, and any two of the input images are processed. A proliferation parameter calculation step for calculating a proliferation parameter representing a change in the amount of the cells per unit time in a predetermined counting region of the image, and an inflow of the cells per unit time of the counting region of each image. Alternatively, a migration parameter calculation step for calculating the migration parameter representing the outflow amount, a region size calculation step for calculating the size of the counting area based on the past proliferation parameter and the past migration parameter, and the region size calculation. It is a cell image processing method including a growth rate calculation step of calculating a cell growth rate based on the growth parameter calculated for the counting area of the size calculated by the step and the migration parameter.

Further, in another aspect of the present invention, a plurality of images acquired by photographing cells at time intervals are input, and any two of the input images are processed. A proliferation parameter calculation step for calculating a proliferation parameter representing a change in the amount of the cells per unit time in a predetermined counting region of the image, and an inflow of the cells per unit time of the counting region of each image. Alternatively, a migration parameter calculation step for calculating the migration parameter representing the outflow amount, a region size calculation step for calculating the size of the counting area based on the past proliferation parameter and the past migration parameter, and the region size calculation. It is a cell image processing program that causes a computer to execute a growth rate calculation step of calculating a cell growth rate based on the growth parameter calculated for the counting area of the size calculated by the step and the migration parameter.

According to the present invention, there is an effect that cell proliferation can be detected accurately and cell culture can be controlled accurately.

It is an overall block diagram which shows the cell image processing apparatus which concerns on one Embodiment of this invention. It is a figure explaining the operation of the cell number change rate calculation part in the cell image processing apparatus of FIG. It is a figure explaining the operation of the cell inflow rate calculation part and the cell outflow rate calculation part in the cell image processing apparatus of FIG. It is a figure explaining the operation of the growth rate calculation part in the cell image processing apparatus of FIG. FIG. 3 is a diagram illustrating an example of a threshold value for determining cell inflow or cell outflow in FIG. It is a flowchart explaining the image processing method of the cell using the cell image processing apparatus of FIG. It is a figure explaining the processing by the modification of the cell image processing apparatus of FIG. It is an overall block diagram which shows the cell image processing apparatus of FIG. It is a flowchart explaining the image processing method of the cell using the cell image processing apparatus of FIG.

The cell image processing apparatus 1, the cell image processing method, and the cell image processing program according to the embodiment of the present invention will be described below with reference to the drawings.
As shown in FIG. 1, in the cell image processing apparatus 1 according to the present embodiment, a plurality of images P _n , P _{n + 1} acquired by photographing at intervals by an imaging unit 50 such as a microscope are input. This is a device that outputs information such as the proliferation rate of cell X to a display unit 60 such as a monitor.

The cell image processing device 1 stores the images P _n and P _{n + 1} input from the imaging unit 50 in the memory 2 and the memory 2 in which the images P _n and P _{n + 1} are sequentially stored in association with the time information at which the images P _n and P _{n + 1} were captured. It is provided with a processing unit 3 for processing the image.
The processing unit 3 processes the images P _n and P _{n + 1} stored in the memory 2, and changes the number of cells per unit time in a predetermined counting area R in the images (evaluation image) P _n and P _{n + 1} . Per unit time into the counting area R in the cell number change rate calculation unit (proliferation parameter calculation unit) 4 for calculating the cell number change rate (proliferation parameter), which is the rate, and the images (evaluation image) P _n , P _{n + 1} . From the cell inflow rate calculation unit (migration parameter calculation unit) 5 that calculates the cell inflow rate (migration parameter), which is the number of inflows of cells X, and the counting region R in the images (evaluation image) P _n , P _{n + 1} . Cell outflow rate calculation unit (migration parameter calculation unit) 6 for calculating cell outflow rate (migration parameter), which is the number of cell X outflow per unit time, and calculated cell number change rate, cell inflow rate, and cell outflow rate. It is provided with a proliferation rate calculation unit 7 for calculating the cell proliferation rate in the counting region R in the proliferation measurement image.
Further, the cell image processing device 1 according to the present embodiment includes a region size calculation unit 8 for setting the size of the counting region R.

As shown in FIG. 2, the cell number change rate calculation unit 4 has two images P _n , acquired at intervals of the first time Δt ₁ , and the number of cells N _n , in the counting region R in P _{n + 1} . the n _{n + 1} counted respectively, each image _{P n,} counted cell number _n n in _{P n + 1,} by dividing the difference between _{n n + 1} in the first hour .DELTA.t1, per unit time the number of cells _{n n,} the _{n n + 1} The cell number change rate ΔN / Δt, which is the rate of change, is calculated.

Cellular influx rate calculating unit 5, as shown in FIG. 3, images P _n on the basis of the image P _n of the two obtained at a just time interval second time _.DELTA.t2, to P n + _1, the count in the P n + ₁ The number of inflows of cells X that entered the region R across the boundary from the adjacent region ΔN _{MOVEINn + 1} was counted, and the counted number of inflows of cells X ΔN _{MOVEINn + 1} was divided by the second time Δt2 to per unit time. The cell inflow rate ΔN _MOVEIN / Δt2, which is the number of inflows of cells X ΔN _MOVEIN , is calculated.

Cells outflow rate calculation part 6 also, as shown in FIG. 3, the second time Δt2 only the image P _n of the two that was acquired at time _intervals, P n + ₁ images P _n on the basis of the _counting of the P n + ₁ The number of outflows of cells X ΔN _{MOVEOUTn + 1} that exited from the region R across the boundary to the adjacent region was counted, and the counted number of outflows of cells X ΔN _{MOVEOUTn + 1} was divided by the second time Δt2 to obtain cells per unit time. The cell outflow rate ΔN _MOVEOUT / Δt2, which is the outflow number ΔN _MOVEOUT of X, is calculated.

The second time Δt2 for calculating the inflow of the cell X into the counting area R and the outflow from the counting area R preferably satisfies the conditional expression (1) below.
Δt2 ≦ D / V (1)
Here, D is the length of cell X, and V is the migration rate of cell X.

The length D and the migration speed V of the cells X may be set to reference values at the time of past counting, or may be set to average values in the counting area R. Further, as shown in FIG. 5, using the threshold value E having a length for determining entry or exit, if cells X enter from the boundary between the counting area R and the adjacent area by the threshold value E, it is determined that the cell X has entered, and from the boundary. When the cell X exits from the counting region R to the adjacent region by the threshold value E, it may be determined to exit.

The region size calculation unit 8 is connected to a memory 2 that stores the past cell proliferation rate ΔN _BIRTH / Δt, the past cell influx rate ΔN _MOVEIN / Δt, and the past cell outflow rate ΔN _MOVEOUT / Δt calculated for each cell X. Based on the past cell proliferation rate ΔN _BIRTH / Δt, cell influx rate ΔN _MOVEIN / Δt, and cell outflow rate ΔN _MOVEOUT / Δt stored in the memory 2, the index of error evaluation is set to a predetermined tolerance Ka or less. The size of the counting area R is determined so as to be.

Specifically, the following conditional expression is satisfied.
ΔN _BIRTH / Δt = A × L ² (2)
(ΔN _{MOVEIN −} ΔN _MOVEOUT ) / Δt = 4 × L × ( _BC ) (3)
_{K = ((ΔN MOVEIN -ΔN MOVEOUT} ) / Δt) / (ΔN BIRTH / Δt)
= 4 (BC) / A × L (4)
here,
A is the number of cells generated in a unit time per unit area in the counting area R.
B is the number of cells that enter the counting area R at the boundary line of the counting area R in a unit time per unit length.
C is the number of cells exiting the counting area R at the boundary line of the counting area R in a unit time per unit length.
L is the length of one side of the counting area R composed of squares.

The area size calculation unit 8 uses the following conditional expression (5) obtained by modifying the conditional expression (4) with the counting tolerance as Ka, and uses the following conditional expression (5) to obtain the length L (size) of one side of the counting area R. Is to be calculated.
L = 4 (BC) / A × Ka (5)

The memory 2 stores the past cell numbers A, B, C, and the counting tolerance Ka, which represent the past cell proliferation rate ΔN _BIRTH / Δt calculated in the past or determined for each cell X.
The region size calculation unit 8 reads the past cell numbers A, B, C and the margin of error Ka from the memory 2, and uses the read cell numbers A, B, C and the margin of error Ka to formulate the conditional expression (5). It is used to calculate the length of one side of the counting area R.

As shown in FIG. 4, the growth rate calculation unit 7 calculates the cell growth rate ΔN _BIRTH / Δt, which is the number of cell divisions per unit time in the counting region R. Specifically, by subtracting the cell inflow rate ΔN _MOVEIN / Δt from the cell number change rate ΔN / Δt and adding the cell outflow rate ΔN _MOVEOUT / Δt by the following equation (6), the cell proliferation rate ΔN _BIRTH / Δt Is to be calculated.
ΔN _BIRTH / Δt = ΔN / Δt−ΔN _MOVEIN / Δt + ΔN _MOVEOUT / Δt (6)

The cell image processing method using the cell image processing apparatus 1 according to the present embodiment configured as described above will be described below.
Here, the cell image processing method according to the present embodiment is described.
Δt1 = Δt2 = Δt
It is explained as.

First, as shown in FIG. 6, the start time t1 of the count, the end time t _m and the time interval Δt of the counting is set (step S1).
Next, the number of cells A, B, C and the margin of error Ka are read from the memory 2 (step S2). Then, the length L of one side of the counting region R is calculated using the read cell numbers A, B, C and the margin of error Ka (region size calculation step S3).

Next, the counter n = 1 is set (step S4), the image P _n acquired at time t _n is processed by the cell number change rate calculation unit 4, and within the square counting area R having a side length L. The number of cells N _n of the cells is counted (proliferation parameter calculation step S5). Further, the image P _{n + 1} obtained cell number N _{n + 1} in the counting region R is processed in the cell number change rate calculating section 4 is counted at time t _{n + 1} from the time t _n has passed by the time interval Delta] t (growth parameters Calculation step S6).

Next, the number of cells ΔN _{MOVEIN n + 1} that entered the counting area R from outside the counting area R and the number of cells that exited from the counting area R to the outside of the counting area R by comparing the image P _n and the image P _{n + 1} with each other. ΔN _{MOVEOUTn + 1} is calculated (migration parameter calculation steps S7 and S8).
Then, in the growth rate calculation unit 7, the cell growth rate ΔN _{BIRTHn + 1} / Δt per unit time is calculated by the following formula (7) (growth rate calculation step S9).
ΔN _{BIRTHn + 1} / Δt
= (N _{n + 1} −N _n ) / Δt−ΔN _{MOVEINn + 1} / Δt + ΔN _{MOVEOUTn + 1} / Δt (7)

Here, the cell number change rate ΔN / Δt, the cell inflow rate ΔN _{MOVEINn + 1} / Δt, the cell outflow rate ΔN _{MOVEOUTn + 1} / Δt, and the cell proliferation rate ΔN _{BIRTHn + 1} / Δt are all calculated based on the same adjacent image. Δt exists in the denominator in all the terms of the above equation (7). Therefore, all the terms can be multiplied by Δt and converted equivalently to the following conditional expression (8).
ΔN _{BIRTHn + 1} = (N _{n + 1} −N _n ) −ΔN _{MOVEINn + 1} + ΔN _{MOVEOUTn + 1} (8)

The counted number of cells N _{n + 1} and the calculated number of proliferating cells ΔN _{BIRTHn + 1} are output to the display unit 60 and displayed as a graph showing the time change with time on the horizontal axis (step S10). At the same time, it is determined whether or not the counter n is n = m (step S11), and if n <m, n is incremented and the process from step S5 is repeated (step S12).

According to the image processing method for the cell image processing apparatus 1 and the cells according to the present embodiment configured as described above, the cell proliferation rate ΔN _{BIRTHn + 1} / Δt is two images P _n , P _{n + 1} with a time interval Δt. Since it is obtained by considering not only the cell number change rate ΔN / Δt but also the cell influx rate ΔN _{MOVEINn + 1} / Δt and the cell outflow rate ΔN _{MOVEOUTn + 1} / Δt, the degree of cell X proliferation due to cell division can be detected more accurately. Therefore, there is an advantage that the cell culture can be managed accurately.

Further, in the present embodiment, the second time Δt2 for calculating the cell inflow rate ΔN _{MOVEINn + 1} / Δt2 and the cell outflow rate ΔN _{MOVEOUTn + 1} / Δt2 is set in the counting region R by satisfying the conditional expression (1). The entering cell X and the exiting cell X can be properly detected. That is, if Δt2 is too short, it is necessary to calculate the cell inflow rate ΔN _{MOVEINn + 1} / Δt2 and the cell outflow rate ΔN _{MOVEOUTn + 1} / Δt2 for a large number of images P _n and P _{n + 1} , which increases the amount of data and increases the burden of counting. On the other hand, if Δt2 is too long, the migration of cell X cannot be captured, and the cell influx rate ΔN _{MOVEINn + 1} / Δt2 and the cell outflow rate ΔN _{MOVEOUTn + 1} / Δt2 cannot be calculated. However, such a condition (1) is satisfied. There is no inconvenience.

In the description of the cell image processing apparatus 1 according to the present embodiment, Δt1 = Δt2 = Δt and the first region = the second region = the counting region R have been described, but they may be different. In this case, the time Δt1 for calculating the cell number change rate ΔN / Δt1 is set to be the shortest, and the time Δt2 for calculating the cell influx rate ΔN _{MOVEINn + 1} / Δt2 and the cell outflow rate ΔN _{MOVEOUTn + 1} / Δt2 is larger than the time Δt1. It is preferable to set an appropriate time that satisfies the conditional expression (1).

Further, in the present embodiment, the processing unit 3 includes a cell number change rate calculation unit 4, a cell inflow rate calculation unit 5, a cell outflow rate calculation unit 6, a growth rate calculation unit 7, and a region size calculation unit 8. , The processing unit 3 may be configured by an electric circuit, or may be configured by one or more processors.
When configured by a processor, the present invention comprises a cell image processing program capable of executing the proliferation parameter calculation steps S5 and S6, the migration parameter calculation steps S7 and S8, the proliferation rate calculation step S9 and the region size calculation step S3 by a computer. You may.

Further, in the present embodiment, as shown in FIGS. 7 to 9, a cell mortality rate calculation unit that calculates the cell mortality rate ΔN _{DEATHn + 1} / Δt, which is the number of deaths of cells X per unit time in the counting region R. 9 may be further provided.
In this case, the growth rate calculation unit 7 may calculate the cell growth rate ΔN _{BIRTHn + 1} / Δt by the following formula (9) (step S13).
ΔN _{BIRTHn + 1} / Δt
= (N _{n + 1} −N _n ) / Δt−ΔN _{MOVEINn + 1} / Δt
+ ΔN _{MOVEOUTn + 1} / Δt + ΔN _{DEATHn + 1} / Δt (9)

By doing so, even in an environment where an event in which cell X dies occurs, the degree of proliferation of cell X due to cell division can be detected more accurately, and cell culture can be managed accurately. There is.
Here, when observing cells X that adhere to the bottom surface of the culture vessel and proliferate as cells X, the number of deaths ΔN _{BIRTHn + 1} of the cells X counts the number of cells N _{n + 1} floating without being able to adhere to the bottom surface. By doing so, it can be easily detected.

Further, in the present embodiment, the time change of the counted number of cells N and the calculated number of proliferating cells ΔN _BIRTH is graphically displayed on the display unit 60, but instead of or in addition to this. _Therefore , at least one of the number of deaths of cell X ΔN _DEATH , the number of cell inflows ΔN _MOVEIN, and the number of cell X outflows ΔN _MOVEOUT may be displayed in a graph.

Further, in the present embodiment, the cell proliferation rate ΔN _{BIRTHn + 1} / Δt is exemplified as the proliferation parameter, but instead, the rate of change in confluency, which is the ratio of the area of the cell region to the area of the counting region R, is adopted. You may.
Further, as the migration parameter, the difference between the cell inflow rate ΔN _{MOVEINn + 1} / Δt and the cell outflow rate ΔN _{MOVEOUTn + 1} / Δt was illustrated, but instead of this, the two images P _n and P _{n + 1} in which the counting region R was photographed The amount of image deformation, which is a conversion parameter of the projective transformation, or the amount of movement of attention cells, which is the average value of the amount of movement of some cells of interest detected by image processing, may be adopted.

Further, in the present embodiment, the size of the counting area R is calculated based on the past proliferation parameters, the migration parameters, and the tolerance Ka, but when the size of the counting area R is predetermined. In addition, the magnitude of the error may be estimated.

Further, in the present embodiment, the length L of one side of the square counting area R is calculated as the size of the counting area R, but instead, when the counting area R is circular, the radius r The length of may be calculated by the following equation (10).
r = 2 × (BC) / (A × K) (10)

Further, the short side L2 of the rectangular counting area R having the long side L1 and the short side L2 may be calculated by the following equation (11) by defining, for example, the long side L1.
L2 = 2 × (BC) × L1 / (A × L1 × K + 2 × (CB)) (11)

1 Cell image processing device 4 Cell number change rate calculation unit (proliferation parameter calculation unit)
5 Cell influx rate calculation unit (migration parameter calculation unit)
6 Cell outflow rate calculation unit (migration parameter calculation unit)
7 Proliferation rate calculation unit (proliferation rate calculation unit)
X cell N _n , N _{n + 1} cell number ΔN / Δt cell number change rate ΔN _MOVEOUT / Δt, ΔN _MOVEIN / Δt2 cell influx rate (migration parameter)
ΔN _MOVEOUT / Δt, ΔN _MOVEOUT / Δt2 Cell outflow rate (migration parameter)
ΔN _BIRTH / Δt cell proliferation rate (proliferation parameter)
P _n , P _{n + 1} image (evaluation image)
S3 Area size calculation step S5, S6 Proliferation parameter calculation step S7, S8 Migration parameter calculation step S9 Proliferation rate calculation step

In the above aspect, the growth parameter calculation unit may calculate the number of cells increased in the counting region per unit time as the growth parameter.
By doing so, it is easy to process the two evaluation images acquired at time intervals, count the number of cells existing in each counting area, and divide the difference by the time interval. Proliferation parameters can be calculated.
Further, in the above aspect, the growth parameter calculation unit may calculate the rate of change in confluency, which is the ratio of the area of the cell region to the area of the counting region, as the growth parameter.

Further, in the above aspect, the migration parameter calculation unit may calculate the difference between the number of cells flowing in from the outer edge of the counting region and the number of cells flowing out from the outer edge of the counting region per unit time as the migration parameter.
By doing so, the two evaluation images acquired at time intervals are processed to count the number of cells that have flowed in and out of the outer edge of each counting area, and the difference is timed. The migration parameter can be easily calculated by dividing by the interval.
Further, in the above aspect, the migration parameter calculation unit may calculate the image deformation amount, which is a conversion parameter of the projection conversion of the two images captured in the counting area, as the migration parameter.
Further, in the above aspect, the migration parameter may use the attention cell migration amount, which is the average value of the migration amounts of some attention cells detected by the image processing, as the migration parameter.
Further, in the above aspect, the image taken by the migration parameter calculation unit at intervals equal to or less than the time obtained by dividing the length of the cell by the speed of the cell may be used.

Further, in the above aspect, a cell mortality rate calculation unit for calculating the cell mortality rate, which is the number of deaths of the cells per unit time in the counting region, may be provided.
Further, in the above aspect, the cell mortality rate may be detected by counting the number of cells suspended in the region per unit time.
Further, in the above aspect, at least one of the proliferation parameter, the migration parameter, the cell proliferation rate and the cell mortality rate may be output as a graph.
Further, in the above aspect, at least one of the proliferation parameter, the migration parameter, the cell proliferation rate and the cell mortality rate may be output as a graph showing the time change.

Claims (8)

Multiple images obtained by photographing cells at time intervals are input, and
Proliferation parameter calculation unit that processes any two of the input evaluation images and calculates a growth parameter representing a change in the amount of the cells per unit time in a predetermined counting region of the evaluation image. When,
A migration parameter calculation unit that calculates a migration parameter representing the inflow or outflow of the cells per unit time of the counting region of each evaluation image, and
A region size calculation unit that calculates the size of the counting region based on the past proliferation parameters and the past migration parameters.
A cell image processing apparatus including a growth rate calculation unit that calculates a cell growth rate based on the growth parameter calculated for the counting area of the size calculated by the region size calculation unit and the migration parameter. The cell image processing apparatus according to claim 1, wherein the growth parameter calculation unit calculates the number of cells increased in the counting region per unit time as the growth parameter. The cell image processing apparatus according to claim 1 or 2, wherein the migration parameter calculation unit calculates the difference between the number of cells flowing in from the outer edge of the counting region and the number of cells flowing out from the outer edge of the counting region as the migration parameter. .. Claims 1 to 3 in which the region size calculation unit calculates a lower limit of the size of the counting region by comparing the ratio of the past migration parameters to the past proliferation parameters with a predetermined margin of error. The cell image processing apparatus according to any one. The cell image processing apparatus according to any one of claims 1 to 4, wherein the counting area is rectangular. It is equipped with a processor that inputs multiple images acquired by photographing cells at time intervals.
The processor has a proliferation parameter representing a change in the amount of the cells per unit time in a predetermined counting region in the past and a migration parameter representing the inflow or outflow of the cells per unit time in the counting region in the past. Based on this, the size of the counting area is calculated, and using the counting area of the calculated size, any two of the input images are processed to perform the counting of the images. A cell image processing apparatus that calculates the proliferation parameter and the migration parameter in the region. Multiple images obtained by photographing cells at time intervals are input, and
A growth parameter calculation step of processing any two of the input images to calculate a growth parameter representing a change in the amount of the cells per unit time within a predetermined counting region of the image. ,
A migration parameter calculation step for calculating a migration parameter representing the inflow or outflow of the cells per unit time of the counting region of each of the images, and
A region size calculation step for calculating the size of the counting region based on the past proliferation parameters and the past migration parameters, and
A cell image processing method including a growth rate calculation step of calculating a cell growth rate based on the growth parameter calculated for the counting area of the size calculated by the region size calculation step and the migration parameter. Multiple images obtained by photographing cells at time intervals are input, and
A growth parameter calculation step of processing any two of the input images to calculate a growth parameter representing a change in the amount of the cells per unit time within a predetermined counting region of the image. ,
A migration parameter calculation step for calculating a migration parameter representing the inflow or outflow of the cells per unit time of the counting region of each of the images, and
A region size calculation step for calculating the size of the counting region based on the past proliferation parameters and the past migration parameters, and
A cell image processing program that causes a computer to execute a growth rate calculation step of calculating a cell growth rate based on the growth parameter calculated for the counting area and the migration parameter of the size calculated by the area size calculation step. ..

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