TWI704335B - Automatic cell analysis device and operation method thereof - Google Patents

Abstract

The present disclosure provides an automatic cell analysis device that includes a camera, an electric stage, a processor and a storage device. The electric stage carries a sample of cells to be tested. The processor commands that the electric stage sequentially moves to a plurality of positions on a predetermined trajectory, so that the camera captures a plurality of pictures corresponding to the plurality of positions. The storage device stores a plurality of pictures. The processor analyzes the number of repeated cells in the overlapping region in any two adjacent pictures in the plurality of pictures, and then subtracts the number of repeated cells from the total number of cells in the plurality of pictures to obtain the cell counting result.

Description

Automated cell analysis device and its operation method

The present invention relates to a device and method, and more particularly to an automated cell analysis device and its operation method.

After the blood sample is stained by fluorescence, the required CTC cells will be separated first. In the process of automated shooting, due to the difference in the area of the glass slide and the shooting field of view (magnification, the size of the photosensitive element), it is generally necessary to shoot dozens of images at different locations to take a full-field blood sample image.

However, if the slide is moved multiple times with an electromechanical stage, it will cause movement accuracy problems, resulting in errors in calculating cells in adjacent overlapping regions. The current technology manually recognizes which cells are duplicated in the overlapping area, but this method is time-consuming and the accuracy varies from person to person.

The present invention proposes an automated cell analysis device and an operation method thereof to improve the problems of the prior art.

In an embodiment of the present invention, the automated cell analysis device provided by the present invention includes a camera, a motorized stage, a processor, and a storage device. Set. The motorized stage carries the cell sample to be tested. The processor causes the electric stage to sequentially move to a plurality of positions on the preset trajectory, so that the camera takes a plurality of pictures corresponding to the plurality of positions. The storage device stores multiple pictures. The processor analyzes the number of repeated cells in the overlapping area of any two adjacent pictures in the plurality of pictures, and then subtracts the number of repeated cells from the total number of cells in the plurality of pictures to obtain a cell count result.

In an embodiment of the present invention, the processor matches all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, and the processor further determines that each of any two adjacent pictures in the plurality of pictures Whether all cells in the overlapping region of has at least one set of matching cell pairs, wherein all matching cell pairs in the at least one set of matching cell pairs meet at least one of the same slope matching or the same distance matching.

In an embodiment of the present invention, when there is no at least one set of matching cell pairs in all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, the processor determines that the number of repeated cells is zero.

In an embodiment of the present invention, when there is only a unique set of matching cell pairs among all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, the processor will set a unique set of matching cells The number of matching cell pairs in the pair is regarded as the number of repeated cells.

In an embodiment of the present invention, when all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures have a complex array of matching cell pairs, the processor selects from the complex array of matching cell pairs with the most matching A group of matched cell pairs of cell pairs, the processor judges whether there are plural one-to-many matched cell pairs in the selected group of matched cell pairs, and if so, matches plural-to-many pairs The cell pair type similarity screening is used to screen out matching cell pairs that are all one-to-one, and the processor uses the number of matching cell pairs in the set of matching cell pairs that have been screened as the number of repeated cells.

In an embodiment of the present invention, in the method of operating an automated cell analysis device provided by the present invention, the automated cell analysis device includes a camera and an electric stage, the electric stage carries the cell sample to be tested, and the operation method includes: The stage moves to a plurality of positions on the preset trajectory in sequence, so that the camera takes a plurality of pictures corresponding to the plurality of positions; analyzes the number of repeated cells in the overlapping area in any two adjacent pictures in the plurality of pictures; and The total number of cells in the multiple pictures minus the number of duplicate cells to get the cell count result.

In an embodiment of the present invention, the operation method further includes: matching all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, and then determining whether any two adjacent pictures in the plurality of pictures Whether all the cells in the respective overlapping regions have at least one set of matching cell pairs, wherein all matching cell pairs in the at least one set of matching cell pairs meet at least one of the same slope matching or the same distance matching.

In an embodiment of the present invention, the operating method further includes: when there is no at least one set of matching cell pairs in all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, determining that the number of repeated cells is zero.

In an embodiment of the present invention, the operating method further includes: when there is only a unique set of matching cell pairs among all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, the only one The number of matched cell pairs in the group matched cell pairs is regarded as the number of repeated cells.

In an embodiment of the present invention, the operation method further includes: When all the cells in the overlapping regions of any two adjacent pictures in the several pictures have a complex array of matching cell pairs, select the group of matching cell pairs with the most matching cell pairs from the complex array of matching cell pairs; judge that it has been selected Whether there are multiple one-to-many matching cell pairs in the group of matching cell pairs, if so, screen the multiple-to-many matching cell pairs for type similarity to screen out matching cell pairs that are all one-to-one; and The number of matched cell pairs in the group of matched cell pairs that have been screened is regarded as the number of repeated cells.

In summary, the technical solution of the present invention has obvious advantages and beneficial effects compared with the prior art. The technical scheme of the present invention makes up for the problem of hardware movement accuracy, does not require manual interpretation, and is time-saving and accurate.

Hereinafter, the above description will be described in detail by way of implementation, and a further explanation will be provided for the technical solution of the present invention.

In order to make the above and other objects, features, advantages and embodiments of the present invention more obvious and understandable, the description of the attached symbols is as follows:

1~7‧‧‧Cell

100‧‧‧Automated cell analysis device

110‧‧‧Camera

120‧‧‧Electric stage

130‧‧‧Processor

140‧‧‧Storage device

210、220‧‧‧Picture

230‧‧‧Overlapping area

310、320‧‧‧Overlapping area

400‧‧‧Movement error range

500‧‧‧Operation method

S501~S511‧‧‧Step

A~G‧‧‧Cell

In order to make the above and other objectives, features, advantages and embodiments of the present invention more comprehensible, the description of the accompanying drawings is as follows: Figure 1 is a block diagram of an automated cell analysis device according to an embodiment of the present invention; And Figure 2 is a schematic diagram of overlapping regions in two adjacent pictures according to an embodiment of the present invention; Figure 3 is a schematic diagram of respective overlapping regions in two adjacent pictures according to an embodiment of the present invention; and 4A, 4B and 4C are schematic diagrams of the movement error range according to an embodiment of the present invention; and FIG. 5 is a flowchart of an operation method of an automated cell analysis device according to an embodiment of the present invention.

In order to make the description of the present invention more detailed and complete, please refer to the attached drawings and various embodiments described below. The same numbers in the drawings represent the same or similar elements. On the other hand, well-known elements and steps are not described in the embodiments to avoid unnecessary limitations on the present invention.

In the implementation and the scope of the patent application, the description of "connection" can generally refer to a component that is indirectly coupled to another component through other components, or that one component is directly connected to another component without other components.

In the scope of implementation and patent application, the description of "connection" can generally refer to a component that is indirectly connected to another component through other components, or that a component is physically connected to another component without going through other components. .

In the implementation mode and the scope of the patent application, unless the article is specifically limited in the context, "a" and "the" can generally refer to a single or plural.

"About", "approximately" or "approximately" used in this article are used to modify any amount that can be slightly changed, but such slight changes will not change its essence. If there is no special description in the implementation, it means that the error range of the value modified with "about", "approximately" or "approximately" is generally allowed within 20%, preferably within 10% Within, and better within five percent.

FIG. 1 is a block diagram of an automated cell analysis device 100 according to an embodiment of the invention. As shown in FIG. 1, the automated cell analysis device 100 may include a camera 110, a motorized stage 120, a processor 130 and a storage device 140. In terms of architecture, the processor 130 is electrically connected to the camera 110, the electric stage 120 and the storage device 140. For example, the processor 130 may be an image processor, a central processing unit, a microcontroller, other processing circuits, or a combination of the foregoing, the storage device 140 may be a hard disk, flash memory or other storage media, and the camera 110 may It is a digital video camera.

The motorized stage 120 can be a device for placing glass slides in a microscope system. The wave plate has a cell sample (such as a blood sample) to be tested, and passes through the biaxial ball screw, linear slide, stepping motor, and processor 130. Under software control, the wave plate can automatically move to various positions according to the set trajectory. Since the camera 110 cannot take a picture of the entire blood sample magnified by the microscope at one time, the processor 130 sets the moving track of the electric stage 120 to move the position of the slide, and takes pictures of each position to ensure that all cells are Taken.

Because the movement of the electric stage 120 has errors (such as: 20μm), and the accuracy of the movement of the stage (such as: 2.5μm) is unstable, these factors may cause the same cell to be repeated in different fields of view It is taken, so the cells in the adjacent area of the picture may be the corresponding duplicate cells, causing the error of the final counting result.

In view of this, please refer to FIGS. 1 and 2 at the same time. FIG. 2 is a schematic diagram of the overlapping area 230 in two adjacent pictures 210 and 220 according to an embodiment of the present invention. When the automated cell analysis device 100 is operating, the motorized stage 120 carries a cell sample to be tested (such as a blood sample in a wave plate). Processor 130 The electric stage 130 is sequentially moved to a plurality of positions on the preset track, so that the camera 110 takes a plurality of pictures 210 and 220 corresponding to the plurality of positions. The storage device 140 stores a plurality of pictures 210 and 220. The processor 130 analyzes the number of repeated cells in the overlapping area 230 in any two adjacent pictures 210 and 220 in the plurality of pictures 210 and 220, and then subtracts the number of repeated cells from the total number of cells in the plurality of pictures 210 and 220 to obtain Result of cell count. This compensates for the problem of the movement accuracy of the electric stage 120, and does not require manual interpretation, which saves time and is accurate.

It should be understood that only two pictures 210 and 220 are shown in Figure 2 for concise description. In practice, in order to realize automatic shooting with a full field of view, those who are familiar with this technique should consider the moving stroke of the electric stage 120 (eg: 40 x 60mm) and the movement track to determine the number of pictures.

In order to further explain the calculation method of the number of repeated cells, please refer to FIGS. 1 to 3 at the same time. FIG. 3 is a schematic diagram of overlapping regions 310 and 320 in two adjacent pictures according to an embodiment of the present invention. It should be understood that the overlapping area 310 of the picture 210 and the overlapping area 320 of the picture 220 become the overlapping area 230 in the second figure. In practice, the size of the overlapping areas 310 and 320 depends on the movement error and movement accuracy of the electric stage 120. If the movement error of the electric stage 120 is larger and or the movement accuracy is lower, the overlap area 310, 320 is larger; on the contrary, if the movement error of the electric stage 120 is smaller or the movement accuracy is higher, the overlap area 310, 320 is smaller. The system setter can manually or the processor 130 can automatically set the size of the overlapping areas 310 and 320 according to the movement error and movement accuracy of the electric stage 120.

As shown in Figure 3, there are cells 1-7 in the overlapping area 310, It is shown in white, and there are cells A to G in the overlapping area 320, and it is shown in black. It should be understood that in this embodiment, the white cells and the black cells are only the cells in the overlapping regions 310 and 320, and do not represent the actual colors of the cells. If the cells 1~7 in the overlapping area 310 are to find the duplicate cells corresponding to each other in the self-overlapping area 320, there may be multiple sets of permutations and combinations, and the processor 130 must sort these combinations one by one and select the most likely overlapping cell. One group.

In order to explain the analysis process, please refer to Figures 4A, 4B, and 4C. Figures 4A, 4B, and 4C are schematic diagrams of the movement error range 400 according to an embodiment of the present invention. The movement error range 400 may be the movement error of the electric stage 120 The upper white cells are associated with all the black cells within 400 within the moving error range of the adjacent repeating area. These black cells may be white repeating cells. In Figure 4A, a white cell corresponds to a black cell in the movement error range 400, and a total of one cell pair. In Figure 4B, one white cell corresponds to two black cells in the movement error range 400, a total of two cell pairs. In Figure 4C, two white cells correspond to three black cells in the movement error range 400, for a total of six cell pairs.

Returning to Figure 3, the processor 130 finds that each white cell 1~7 in the overlapping area 310 corresponds to a black cell in its dynamic error range 400. The total result: cell pair 1A (that is, cell 1 corresponds to cell A, the following is Analogy), cell pair 1B, cell pair 2A, cell pair 2B, cell pair 3C, cell pair 3D, cell pair 4D, cell pair 4E, cell pair 4F, cell pair 5D, cell pair 5E, cell pair 5F, cell pair 6E , Cell to 6F, Cell to 6G, Cell to 7G.

〔(x1-x2)^2+(y1-y2)^2〕±q)，其中x1,y1為細胞對中白色細胞的座標值，x2,y2為細胞對中黑色細胞的座標值，p為允許傾斜誤差(如：0.02)，q為允許距離誤差(如：20)。處理器130把這些數據優先傾斜度後距離的方案進行排序分組，拆分出來的隊列如下： Next, the processor 130 groups the filtered total results, and calculates the corresponding relationship between the center points of all adjacent cells in the plane coordinate system, such as: inclination=(y2-y1)/(x2-x1)±p and Distance=(

[(X1-x2)^2+(y1-y2)^2]±q), where x1, y1 are the coordinate values of the white cells in the cell pair, x2, y2 are the coordinate values of the black cells in the cell pair, and p is Allowable tilt error (such as: 0.02), q is the allowable distance error (such as: 20). The processor 130 sorts and groups these data prioritized inclination distance solutions, and the split queues are as follows:

The first group: matched cell pair 1A, matched cell pair 2B, matched cell pair 3D, matched cell pair 4E, matched cell pair 4F, matched cell pair 5E, matched cell pair 5F, matched cell pair 6G.

The second group: matched cell pair 2A, matched cell pair 4D, matched cell pair 5D, and matched cell pair 7G.

The third group: matched cell pair 3C.

To sum up, the processor 130 matches all the cells in the respective overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 in the plurality of pictures 210 and 220, and the processor 130 further determines the plurality of pictures 210 and 220. Whether there are at least one set of matching cell pairs for all cells in the respective overlapping regions 310, 320 in any two adjacent pictures 210 and 220 in 220, wherein all matching cell pairs in the at least one set of matching cell pairs conform to the same slope At least one of matching or matching at the same distance. For example, the matched cell pairs 1A, 2B, 3D, 4E, 4F, 5E, 5F, and 6G in the first group all match with the same first slope, and the matched cell pairs in the second group 2A, 4D, 5D, 7G All match with the same second slope, and the matched cell pair 3C in the third group meets the third slope match, where the first, second, and third slopes are different from each other.

Next, when all the cells in the overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 in the plurality of pictures 210 and 220 have a complex array of matching cell pairs (such as: the first, second, third When group), the processor 130 selects a group of matched cell pairs with the most matched cell pairs from the complex array of matched cell pairs. For example, the processor 130 regards the first group as the selected group of matching cell pairs (ie, the optimal queue), because the number of the same slope is the most. Normally, the queue of duplicate cells is the group with the most.

The final selected queue may have multiple groups with the same number, or the same cell in the queue may have multiple duplicate cells. In response to this situation, the processor 130 adds morphological combination determination and further filters the results. Specifically, the processor 130 determines whether there are a plurality of one-to-many matching cell pairs (such as 4E, 4F, 5E, 5F) in the selected group of matching cell pairs, and if so, the plurality of pair-to-many matching cells The type similarity screening is performed to screen out matching cell pairs that are all one-to-one, and the processor 130 uses the number of matching cell pairs in the set of matching cell pairs that have been screened as the number of repeated cells.

For example, in the first group: 1A, 2B, 3D, 4E, 4F, 5E, 5F, 6G in the above queue, cells 4 and 5 correspond to the two groups of cells E and F because they are too close. Find each group separately Corresponding to the cell morphological determination coefficient, the closest group is selected as 4E and 5F. The processor 130 sets the group of matched cell pairs that have been screened into 1A, 2B, 3D, 4E, 5F, and 6G, and the number of repeated cells is six.

In an embodiment of the present invention, the morphological determination coefficient used in the morphological similarity screening between cells may be the roundness determination coefficient, the diameter ratio coefficient, the relationship between the eccentricity of the image and the inertia rate, the current cell and Corresponding to repeated fine The average brightness ratio of the cell, the current cell size (number of pixels) and the ratio of the corresponding repeated cells... etc. If the difference between the morphological determination coefficients of two cells is smaller, the similarity of the morphology of the two cells is higher; conversely, if the difference of the morphological determination coefficients of the two cells is larger, the similarity of the morphology of the two cells is lower.

On the other hand, if there is no at least one set of matching cell pairs in all the cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, the processor 130 determines that the number of repeated cells is zero. Or, if there is only a unique set of matching cell pairs in all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, the processor 130 will determine the matching cell pair in the only set of matching cell pairs. The number is taken as the number of repeated cells.

In order to further explain the operation method of the automated cell analysis device 100, please refer to FIGS. 1 to 5 at the same time. FIG. 5 is a flowchart of an operation method 500 of the automated cell analysis device 100 according to an embodiment of the present invention. It should be understood that the operation method 500 is an in vitro cell counting method through the automated cell analysis device 100, and is not a method of diagnosis, treatment or surgery for humans or animals. As shown in Figure 5, the operation method 500 includes steps S501 to S511 (it should be understood that the steps mentioned in this embodiment can be adjusted according to actual needs, except for those whose order is specifically stated. It can even be executed simultaneously or partially simultaneously).

In step S501, the camera 110 starts shooting. In step S502, the electric stage 120 is sequentially moved to a plurality of positions on the preset trajectory, so that the camera 110 takes a plurality of pictures 210 and 220 corresponding to the plurality of positions. In step S503, the storage device 140 stores a plurality of pictures 210 and 220. In step S504, circle all cells in pictures 210 and 220 and Record cell properties.

In step S505, all the cells 1-7 in the overlapping regions 310, 320 of any two adjacent pictures 210, 220 in the plurality of pictures 210, 220 are matched with A~G. In step S506, it is determined whether all the cells in the respective overlapping regions 310, 320 of any two adjacent pictures 210, 220 of the plurality of pictures 210, 220 have at least one set of matching cell pairs, wherein the at least one set of matching cell pairs All matching cell pairs in, meet at least one of the same slope matching or the same distance matching (ie, conditions).

When all the cells 1~7 and A~G in any two adjacent pictures 210 and 220 in the respective overlapping regions 310 and 320 of the plural pictures 210 and 220 do not have at least one group of matching cell pairs (ie, group 0) In step S510, it is determined that the number of repeated cells is zero.

When any two adjacent pictures 210 and 220 in the plurality of pictures 210 and 220, all the cells 1~7 in the respective overlapping regions 310, 320 and A~G only have a unique set of matching cell pairs (ie, 1 Group), in step S510, the number of matching cell pairs in the unique set of matching cell pairs is used as the number of repeated cells.

When all the cells 1~7 in the overlapping regions 310, 320 of any two adjacent pictures 210 and 220 in the plural pictures 210 and 220 have a complex array of matching cell pairs with A~G, in step S507, from the plural The group of matched cell pairs with the most matched cell pairs is selected from the group of matched cell pairs.

In step S508, it is determined whether the selected groups of matching cell pairs (for example, the maximum number of matching cell pairs are the same) or whether there are multiple one-to-many matching cell pairs in the selected group of matching cell pairs. If yes, in step S509, the selected multiple groups or multiple pair-to-many matching cell types in a group State similarity screening to screen out matching cell pairs that are all one-to-one. If there are multiple groups, select the group with the most one-to-one matching cell pairs. In step S510, the number of matched cell pairs in the group of matched cell pairs that have been screened is used as the number of repeated cells.

In step S511, the total number of cells in the plurality of pictures 210 and 220 is subtracted from the number of repeated cells to obtain a cell count result.

In summary, the technical solution of the present invention has obvious advantages and beneficial effects compared with the prior art. The technical scheme of the present invention makes up for the problem of hardware movement accuracy, does not require manual interpretation, and is time-saving and accurate.

Although the present invention has been disclosed in the above embodiments, it is not intended to limit the present invention. Anyone familiar with the art can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection of the present invention The scope shall be subject to those defined in the attached patent scope.

100‧‧‧Automated cell analysis device

110‧‧‧Camera

120‧‧‧Electric stage

130‧‧‧Processor

140‧‧‧Storage device

Claims (8)

An automated cell analysis device, comprising: a camera; an electric stage, which carries a cell sample to be tested; a processor, which makes the electric stage move to a plurality of positions on a preset track in sequence, so that the camera can shoot A plurality of pictures corresponding to the positions; and a storage device for storing the pictures, wherein the processor analyzes the number of a repeated cell in the overlapping area of any two adjacent pictures in the pictures, and then the The total number of cells in the picture is subtracted from the number of repeated cells to obtain a cell count result, wherein the processor matches all the cells in the respective overlapping regions of any two adjacent pictures in the pictures, and the processor further Determine whether there are at least one set of matching cell pairs for all cells in the respective overlapping regions of any two adjacent pictures in the pictures, wherein all matching cell pairs in the at least one set of matching cell pairs conform to the same slope matching or the same At least one of the distance matches. The automated cell analysis device according to claim 1, wherein when there is no at least one set of matching cell pairs in all cells in the respective overlapping regions of any two adjacent pictures in the pictures, the processor determines the duplicate The number of cells is zero. The automated cell analysis device described in claim 1, Wherein, when there is only a unique set of matching cell pairs in all the cells in the respective overlapping regions of any two adjacent pictures in the pictures, the processor will determine the matching cell pair in the unique set of matching cell pairs. The number is used as the number of repeating cells. The automated cell analysis device according to claim 1, wherein when all cells in the overlapping regions of any two adjacent pictures in the pictures have the groups of matching cell pairs, the processor matches from the groups A group of matching cell pairs with the most matching cell pairs is selected among the cell pairs, and the processor determines whether there are a plurality of one-to-many matching cell pairs in the selected group of matching cell pairs, and if so, the pair-to-many matching Cell pair type similarity screening is used to screen out matching cell pairs that are all one-to-one, and the processor uses the number of matching cell pairs in the set of matching cell pairs that have been screened as the number of repeated cells. An operation method of an automated cell analysis device, the automated cell analysis device comprising a camera and an electric stage, the electric stage carrying a cell sample to be tested, the operation method comprising: sequentially moving the electric stage to a preset Set a plurality of positions on the trajectory, so that the camera takes a plurality of pictures corresponding to these positions; match all the cells in the respective overlapping regions of any two adjacent pictures in the pictures, and then determine the pictures Whether there are at least one set of matching cell pairs for all cells in the respective overlapping regions of any two adjacent pictures, wherein all matching cell pairs in the at least one set of matching cell pairs are in line with the same slope matching or the same distance matching At least one Analyze the number of a repeated cell in the overlapping area in any two adjacent pictures in the pictures; and subtract the number of repeated cells from the total number of cells in the pictures to obtain a cell count result. The operation method according to claim 5, further comprising: determining the number of repeated cells when there is no matching cell pair in all cells in the respective overlapping regions of any two adjacent pictures in the pictures Is zero. The operation method described in claim 5 further includes: when there is only a unique set of matching cell pairs among all cells in the respective overlapping regions of any two adjacent pictures in the pictures, the only one The number of matched cell pairs in the set of matched cell pairs is used as the number of repeated cells. The operation method according to claim 5, further comprising: when all cells in the respective overlapping regions of any two adjacent pictures in the pictures have the groups of matching cell pairs, select from the groups of matching cell pairs Select a group of matching cell pairs with the most matching cell pairs; determine whether there are multiple one-to-many matching cell pairs in the selected group of matching cell pairs, and if so, the type similarity of the pair-many matching cell pairs Screening to screen out matching cell pairs that are all one-to-one; and the number of matching cell pairs in the set of matching cell pairs that have been screened is used as the number of repeated cells.

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