CN109709346B - Automated cell analysis device and method of operating the same - Google Patents
Automated cell analysis device and method of operating the same Download PDFInfo
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Abstract
The invention provides an automatic cell analysis device, which comprises a camera, an electric carrying platform, a processor and a storage device. The motorized stage carries a cell sample to be tested. The processor makes the electric carrying platform move to a plurality of positions on a preset track in sequence, so that the camera shoots a plurality of pictures corresponding to the positions. The storage device stores a plurality of pictures. The processor analyzes the number of the repeated cells in the overlapping area of any two adjacent pictures in the multiple pictures, and then subtracts the number of the repeated cells from the total number of the cells in the multiple pictures to obtain a cell counting result.
Description
Technical Field
The present invention relates to an apparatus and method, and more particularly, to an automated cell analysis apparatus and a method of operating the same.
Background
After the blood sample is stained by fluorescence, the desired CTC cells are isolated. In the automatic imaging process, due to the difference in the area of the slide and the imaging field of view (magnification, size of the photosensitive element), dozens of images at different positions are generally required to be imaged to form an image of the blood sample in the full field of view.
However, moving the slide many times with the electromechanical stage creates problems with the accuracy of the movement, resulting in errors in calculating the cells in adjacent overlapping areas. Current techniques manually identify which cells are overlapping in the overlapping area, but this is time consuming and varies from person to person with accuracy.
Disclosure of Invention
The invention provides an automatic cell analysis device and an abdomen sound monitoring device thereof, which solve the problems of the prior art.
In an embodiment of the present invention, an automated cell analysis apparatus includes a camera, a motorized stage, a processor, and a storage device. The motorized stage carries a cell sample to be tested. The processor makes the electric carrying platform move to a plurality of positions on a preset track in sequence, so that the camera shoots a plurality of pictures corresponding to the positions. The storage device stores a plurality of pictures. The processor analyzes the number of the repeated cells in the overlapping area of any two adjacent pictures in the plurality of pictures, and further subtracts the number of the repeated cells from the total number of the cells in the plurality of pictures to obtain a cell counting result.
In an embodiment of the invention, the processor matches all cells in the overlapping region of any two adjacent pictures in the plurality of pictures, and the processor further determines whether all cells in the overlapping region of any two adjacent pictures in the plurality of pictures have at least one set of matched cell pairs, wherein all matched cell pairs in the at least one set of matched cell pairs meet at least one of a same slope match or a same distance match.
In an embodiment of the invention, when there is no at least one set of matched cell pairs in all cells in the overlapping region in any two adjacent pictures in the multiple pictures, the processor determines that the number of the repeated cells is zero.
In an embodiment of the invention, when there is only one set of matched cell pairs in all cells in the overlapping region in any two adjacent pictures in the plurality of pictures, the processor uses the number of matched cell pairs in the only one set of matched cell pairs as the number of the repeated cells.
In an embodiment of the invention, when all cells in the overlapping region of any two adjacent pictures in the multiple pictures have multiple sets of matched cell pairs, the processor selects a set of matched cell pairs having the most matched cell pairs from the multiple sets of matched cell pairs, the processor determines whether the selected set of matched cell pairs has multiple one-to-many matched cell pairs, if so, the multiple pairs of matched cell pairs are subjected to similarity screening to screen out the matched cell pairs which are all one-to-one, and the processor takes the number of the matched cell pairs in the screened set of matched cell pairs as the number of the repeated cells.
In an embodiment of the present invention, in an operating method of an automated cell analyzer, the automated cell analyzer includes a camera and a motorized stage, the motorized stage carries a cell sample to be tested, and the operating method includes: the electric carrying platform is sequentially moved to a plurality of positions on a preset track, so that a camera shoots a plurality of pictures corresponding to the positions; analyzing the number of the repeated cells in the overlapping area of any two adjacent pictures in the plurality of pictures; and subtracting the number of the repeated cells from the total number of the cells in the plurality of pictures to obtain a cell counting result.
In an embodiment of the invention, the operation method further includes: and matching all cells in the overlapping region of any two adjacent pictures in the plurality of pictures, and further judging whether all cells in the overlapping region of any two adjacent pictures in the plurality of pictures have at least one group of matched cell pairs, wherein all the matched cell pairs in at least one group of matched cell pairs meet at least one of the same slope matching or the same distance matching.
In an embodiment of the present invention, the operation method further includes: and when at least one group of matched cell pairs does not exist in all cells in the respective overlapping regions of any two adjacent pictures in the plurality of pictures, judging that the number of the repeated cells is zero.
In an embodiment of the present invention, the operation method further includes: when only one group of matched cell pairs exists in all cells in the overlapping region of any two adjacent pictures in the multiple pictures, taking the number of the matched cell pairs in the unique group as the number of the repeated cells.
In an embodiment of the present invention, the operation method further includes: when all cells in respective overlapping regions of any two adjacent pictures in the plurality of pictures have a plurality of sets of matched cell pairs, selecting a set of matched cell pairs having the most matched cell pairs from the plurality of sets of matched cell pairs; judging whether the selected group matching cell pairs have a plurality of one-to-many matching cell pairs, if so, screening the similarity of the types of the plurality of one-to-many matching cell pairs to screen out the matching cell pairs which are all one-to-one; and taking the number of matched cell pairs in the screened matched cell pair group as the number of the repeated cells.
In conclusion, compared with the prior art, the technical scheme of the invention has obvious advantages and beneficial effects. The technical scheme of the invention makes up the problem of hardware movement precision, does not need manual interpretation, and is time-saving and accurate.
The above description will be described in detail by embodiments, and further explanation will be provided for the technical solution of the present invention.
Drawings
In order to make the aforementioned and other objects, features, advantages and embodiments of the invention more comprehensible, the following description is given:
FIG. 1 is a block diagram of an automated cell analysis apparatus according to one embodiment of the present invention; and
FIG. 2 is a diagram illustrating an overlapping area in two adjacent pictures according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of overlapping areas in two adjacent pictures according to an embodiment of the present invention; and
FIGS. 4A, 4B, and 4C are schematic diagrams of a motion error range according to an embodiment of the invention; and
FIG. 5 is a flow chart of a method of operating an automated cellular analysis device according to an embodiment of the present invention.
Description of the symbols
In order to make the aforementioned and other objects, features, and advantages of the present invention comprehensible, the following description is made:
1-7: cells
100: automated cell analysis device
110: camera with a camera module
120: electric carrying platform
130: processor with a memory having a plurality of memory cells
140: storage device
210. 220, and (2) a step of: picture frame
230: overlapping area
310. 320, and (3) respectively: overlapping area
400: range of motion error
500: method of operation
S501-S511: step (ii) of
A to G: cells
Detailed Description
In order to make the description of the present invention more complete and complete, reference is made to the accompanying drawings and the following description of various embodiments, in which like reference numerals refer to the same or similar elements. In other instances, well-known elements and steps have not been described in detail in order to avoid unnecessarily obscuring the present invention.
In the description of the embodiments and the claims, reference to "connected" may generally mean that an element is indirectly coupled to another element through another element or that an element is directly coupled to another element without passing through the other element.
In the description and claims, references to "connected" may refer broadly to an element being indirectly connected to another element through another element or to an element being physically connected to another element without the aid of another element.
In the embodiments and claims, the terms "a" and "an" can mean "one or more" unless the context specifically states otherwise.
As used herein, "about" or "approximately" is intended to modify the quantity by which any slight variation is made, but such slight variation does not alter the nature thereof. Unless otherwise specified, the range of error for values modified by "about", "about" or "approximately" is generally tolerated within twenty percent, preferably within ten percent, and more preferably within five percent.
FIG. 1 is a block diagram of an automated cellular analysis device 100 according to an embodiment of the present invention. As shown in fig. 1, the automated cell analysis apparatus 100 may include a camera 110, a motorized stage 120, a processor 130, and a storage device 140. In the configuration, the processor 130 is electrically connected to the camera 110, the motorized 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 thereof, the storage device 140 may be a hard disk, a flash memory, or other storage medium, and the camera 110 may be a digital video camera.
The motorized stage 120 can be a device for placing a slide in a microscope system, and the wave plate has a cell sample (e.g., a blood sample) to be tested, and can be automatically moved to various positions according to a predetermined trajectory under the software control of the dual-axis ball screw, linear slide, stepper motor and processor 130. Since the camera 110 cannot capture the whole blood sample at a time after the whole blood sample is magnified by the microscope, the processor 130 sets the position of the slide moving along the movement track of the motorized stage 120, and captures a picture at each position to ensure that all cells are captured.
Because there is an error (e.g., a movement error of 20 μm) in the movement of the motorized stage 120 and the precision of the stage movement (e.g., a movement precision of 2.5 μm) is not stable, and these factors may cause the same cell to be repeatedly captured in different fields of view, cells in adjacent areas of the picture may be corresponding repeated cells, resulting in an error in the final counting result.
Referring to fig. 1-2, fig. 2 is a schematic diagram of an overlapping area 230 in two adjacent pictures 210 and 220 according to an embodiment of the invention. During operation of the automated cellular analysis device 100, the motorized stage 120 carries a sample of cells to be analyzed (e.g., a blood sample in a waveplate). The processor 130 sequentially moves the motorized stage 130 to a plurality of positions on a predetermined trajectory, so that the camera 110 captures a plurality of pictures 210 and 220 corresponding to the plurality of positions. The storage device 140 stores a plurality of pictures 210, 220. The processor 130 analyzes the number of the repeated cells in the overlapping area 230 of any two adjacent pictures 210 and 220 in the plurality of pictures 210 and 220, and subtracts the number of the repeated cells from the total number of the cells in the plurality of pictures 210 and 220 to obtain the cell count result. Therefore, the problem of the moving precision of the electric carrying platform 120 is solved, manual interpretation is not needed, and time is saved and accuracy is achieved.
It should be understood that only two pictures 210 and 220 are shown in fig. 2 for simplicity, and in practice, in order to realize the full-view automatic shooting, a person skilled in the art determines the number of pictures while looking at the moving stroke (e.g., 40 × 60mm) and the moving track of the motorized stage 120.
To further illustrate the calculation of the number of the repeat cells, please refer to fig. 1-3, and fig. 3 is a schematic diagram of the overlap regions 310 and 320 in two adjacent pictures according to an embodiment of the invention. It should be appreciated that the overlapping area 310 of the picture 210 and the overlapping area 320 of the picture 220 are overlapped to form the overlapping area 230 of fig. 2. In practice, the size of the overlapping areas 310 and 320 depends on the movement error and the movement precision of the motorized stage 120. If the movement error of the motorized stage 120 is larger and the movement precision is lower, the overlapping areas 310 and 320 are larger; on the contrary, the smaller the movement error of the motorized stage 120 and/or the higher the movement precision, the smaller the overlapping areas 310 and 320. The system operator can manually set the size of the overlapping areas 310 and 320 or the processor 130 can automatically set the size of the overlapping areas according to the movement error and the movement precision of the motorized stage 120.
As shown in FIG. 3, the overlapping area 310 has cells 1-7, which are depicted as white, and the overlapping area 320 has cells A-G, which are depicted as black. It should be understood that the white cells and the black cells in this embodiment are only for convenience of describing the cells in the respective overlapping regions 310, 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 overlapping cells corresponding to each other in the overlapping area 320, there may be multiple permutation combinations, and the processor 130 should select the most likely overlapping cell from the permutation combinations.
For convenience of illustration, please refer to fig. 4A, 4B, and 4C, fig. 4A, 4B, and 4C are schematic diagrams of a movement error range 400 according to an embodiment of the present invention, where the movement error range 400 can be a movement error range of the motorized stage 120, and the upper white cells are associated with all black cells in the movement error range 400 of the adjacent repeating area, and the black cells may be white repeating cells. In FIG. 4A, a white cell corresponds to a black cell within the motion error range 400, for a single cell pair. In FIG. 4B, one white cell corresponds to two black cells in the range of motion error 400, for a total of two cell pairs. In FIG. 4C, two white cells correspond to three black cells in the range of motion error 400, for a total of six cell pairs.
Returning to fig. 3, the processor 130 finds each of the white cells 1-7 in the overlapped area 310 corresponding to the black cell in the dynamic error range 400, and the total result is: cell pair 1A (i.e., cell 1 corresponds to cell a, and so on hereinafter), cell pair 1B, cell pair 2A, cell pair 2B, cell pair 3C, cell pair 3D, cell pair 4E, cell pair 4F, cell pair 5D, cell pair 5E, cell pair 5F, cell pair 6E, cell pair 6F, cell pair 6G, and cell pair 7G.
Next, the processor 130 groups the total result from the screening, and counts the correspondence of the central points of all neighboring cells in the plane coordinate system, such as: the gradient is (y2-y1)/(x2-x1) ± p and the distance is (√ [ (x1-x2) ^2+ (y1-y2) ^2] ± q), wherein x1 and y1 are coordinate values of white cells in a cell pair, x2 and y2 are coordinate values of black cells in a cell pair, p is an allowable gradient error (e.g.: 0.02), and q is an allowable distance error (e.g.: 20). The processor 130 sorts the data into groups according to the distance-after-gradient scheme, and the split queues are as follows:
a 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.
Second group: match cell pair 2A, match cell pair 4D, match cell pair 5D, match cell pair 7G.
Third group: matched cell pair 3C.
To sum up, the processor 130 matches all the cells in the overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 in the multiple pictures 210 and 220, and the processor 130 further determines whether all the cells in the overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 in the multiple pictures 210 and 220 have at least one set of matched cell pairs, wherein all the matched cell pairs in the at least one set of matched cell pairs meet at least one of the same slope matching or the same distance matching. For example, the matching cell pairs 1A, 2B, 3D, 4E, 4F, 5E, 5F, 6G in the first group all conform to the same first slope match, the matching cell pairs 2A, 4D, 5D, 7G in the second group all conform to the same second slope match, and the matching cell pair 3C in the third group conform to the third slope match, where the first, second, and third slope users are different.
Next, when all cells in the overlapping regions 310, 320 of any two adjacent pictures 210, 220 in the plurality of pictures 210, 220 have multiple sets of matched cell pairs (e.g., the first, second, and third sets described above), the processor 130 selects a set of matched cell pairs having the most matched cell pairs from the multiple sets of matched cell pairs. For example, the processor 130 uses the first set as the selected set of matching cell pairs (i.e., the optimal queue) because the number of identical dips is the greatest. The cohort of duplicate cells is normally the largest group.
The final selected cohort may have multiple groups of the same number, or multiple replicates of the same cell within the cohort. For this case the processor 130 adds a morphological combination decision and further screens the results. Specifically, processor 130 determines whether the selected set of matched cell pairs has a plurality of one-to-many matched cell pairs (e.g., 4E, 4F, 5E, 5F), if yes, the type similarity of the plurality of pairs of matched cell pairs is screened to screen out the matched cell pairs that are all one-to-one, and processor 130 uses the number of the matched cell pairs in the screened set of matched cell pairs as the number of the repeat cells.
For example, the first group: 1A, 2B, 3D, 4E, 4F, 5E, 5F, 6G, the cells 4 and 5 correspond to 2 groups of cells E and F, respectively, because they are too close, the morphological judgment coefficients of the corresponding cells of each group are calculated, and the closest group is selected as 4E and 5F. Processor 130 assigns the selected matched set of pairs of cells to 1A, 2B, 3D, 4E, 5F, 6G, and repeats the number of cells to six.
In an embodiment of the present invention, the morphology determination coefficient used for the inter-cell morphology similarity screening may be a circularity determination coefficient, a diameter ratio coefficient, a relationship between eccentricity and inertia of an image, an average brightness ratio of a current cell and a corresponding repeat cell, a size (number of pixels) of the current cell and a ratio … of the corresponding repeat cell, and the like. If the difference between the shape determination coefficients of the two cells is smaller, the similarity between the shapes of the two cells is higher; on the contrary, if the difference between the two cell morphology determination coefficients is larger, the similarity between the two cell morphologies is lower.
On the other hand, if there is not at least one set of matched cell pairs in all cells in the overlapping region of any two adjacent pictures in the plurality of pictures, the processor 130 determines that the number of the repeated cells is zero. Alternatively, if there is only one set of matched cell pairs in all cells in the overlapping region in any two adjacent pictures in the plurality of pictures, the processor 130 uses the number of matched cell pairs in the only one set of matched cell pairs as the number of the repeated cells.
To further illustrate the operation of the automated cell analysis apparatus 100, please refer to fig. 1-5, and fig. 5 is a flowchart of an operation method 500 of the automated cell analysis apparatus 100 according to an embodiment of the invention. It should be appreciated that the method 500 is an in vitro cell counting method via the automated cell analysis device 100, and is not a human or animal diagnostic, therapeutic, or surgical method. As shown in fig. 5, the operation method 500 includes a step S501 and a step S511 (it should be understood that, except for the specific sequence, the steps mentioned in the present embodiment may be performed simultaneously or partially simultaneously according to the actual requirement.
In step S501, the camera 110 starts shooting. In step S502, the motorized stage 120 is sequentially moved to a plurality of positions on the predetermined trajectory, so that the camera 110 captures 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, all cells are circled in the pictures 210 and 220 and the cell attributes are recorded.
In step S505, all cells 1 to 7 in the overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 of the plurality of pictures 210 and 220 are matched with a to G. In step S506, it is determined whether all cells in the overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 in the multiple pictures 210 and 220 have at least one matched cell pair, wherein all matched cell pairs in the at least one matched cell pair meet at least one of the same slope match or the same distance match (i.e., the condition).
When there is not at least one set of matched cell pairs (i.e., 0 set) in all cells 1 to 7 and a to G in the overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 in the multiple pictures 210 and 220, it is determined in step S510 that the number of the repeated cells is zero.
When there is only one set of matched cell pairs (i.e., 1 set) in all cells 1 to 7 and a to G in the overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 in the plurality of pictures 210 and 220, the number of matched cell pairs in the only one set of matched cell pairs is used as the number of the repeated cells in step S510.
When there are a plurality of sets of matched cell pairs for all the cells 1 to 7 and a to G in the overlapping regions 310 and 320 of any two adjacent pictures 210 and 220 in the plurality of pictures 210 and 220, a set of matched cell pairs having the most matched cell pairs is selected from the plurality of sets of matched cell pairs in step S507.
In step S508, it is determined whether there are multiple one-to-many pairs of matching cells in the selected multiple sets of matching cell pairs (e.g., the number of the most matching cell pairs is the same) or the selected set of matching cell pairs. If yes, in step S509, the similarity of the types of the multiple pairs of matching cells in the selected multiple groups or groups is screened to screen out the pairs of matching cells that are all one-to-one. If there are more groups, the group with the largest number of one-to-one matched cell pairs is selected. In step S510, the number of matched cell pairs in the selected matched cell pair set is used as the number of repeat cells.
In step S511, the total number of cells in the multiple pictures 210 and 220 is subtracted by the number of the repeated cells to obtain a cell count result.
In conclusion, compared with the prior art, the technical scheme of the invention has obvious advantages and beneficial effects. The technical scheme of the invention makes up the problem of hardware movement precision, does not need manual interpretation, and is time-saving and accurate.
Although the present invention has been described with reference to the above embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (8)
1. An automated cellular analysis device, comprising:
a camera;
an electric carrying platform for carrying a cell sample to be tested;
the processor is used for enabling the electric carrying platform to sequentially move to a plurality of positions on a preset track, and enabling the camera to shoot a plurality of pictures corresponding to the positions; and
a storage device for storing the pictures,
wherein the processor analyzes a number of repeat cells in an overlapping region in any two adjacent pictures of the pictures to subtract the number of repeat cells from a total number of cells in the pictures to obtain a cell count result, further comprising: the processor matches all cells in the overlapping area of any two adjacent pictures in the pictures, and the processor further determines whether all cells in the overlapping area of any two adjacent pictures in the pictures have at least one group of matched cell pairs, wherein all the matched cell pairs in the at least one group of matched cell pairs meet at least one of the same slope matching or the same distance matching.
2. The automated cell analysis apparatus of claim 1, wherein the processor determines that the number of duplicate cells is zero when there is no at least one set of matched cell pairs in all cells in the overlapping region of each of any two adjacent pictures in the pictures.
3. The automated cell analysis apparatus of claim 1, wherein the processor uses the number of matched cell pairs in a unique set of matched cell pairs as the number of duplicate cells when there is only a unique set of matched cell pairs in all cells in the overlapping region in each of two adjacent pictures.
4. The automated cell analysis apparatus of claim 1, wherein the processor selects a set of matched cell pairs having the most matched cell pairs from the sets of matched cell pairs when all cells in the overlapping region of any two adjacent ones of the sets of matched cell pairs exist in the overlapping region, the processor determines whether the selected set of matched cell pairs has a plurality of one-to-many matched cell pairs, if so, the pairs are further subjected to similarity screening to screen out the pairs that are all one-to-one matched cell pairs, and the processor uses the number of matched cell pairs in the screened set of matched cell pairs as the number of the overlapping cells.
5. An operation method of an automated cell analysis device, the automated cell analysis device comprising a camera and a motorized stage, the motorized stage carrying a cell sample to be tested, the operation method comprising:
the electric carrying platform is sequentially moved to a plurality of positions on a preset track, so that the camera shoots a plurality of pictures corresponding to the positions;
analyzing the number of repeat cells in the overlapping region of any two adjacent pictures in the pictures; and
subtracting the number of the repeated cells from the total number of the cells in the pictures to obtain a cell counting result;
and the processor further judges whether all cells in the overlapping region of any two adjacent pictures in the pictures have at least one group of matched cell pairs, wherein all the matched cell pairs in the at least one group of matched cell pairs meet at least one of the same slope matching or the same distance matching.
6. The method of operation of claim 5, further comprising:
when all the cells in the overlapping area of any two adjacent pictures in the pictures do not have the at least one group of matched cell pairs, the number of the repeated cells is judged to be zero.
7. The method of operation of claim 5, further comprising:
when only one set of matched cell pairs exists in all cells in the overlapping region of any two adjacent pictures in the pictures, taking the number of the matched cell pairs in the only one set of matched cell pairs as the number of the repeated cells.
8. The method of operation of claim 5, further comprising:
selecting a set of matched cell pairs having a most matched cell pair from the sets of matched cell pairs when all cells in respective overlapping regions of any two adjacent pictures in the sets of matched cell pairs;
judging whether the selected group of matched cell pairs has a plurality of one-to-many matched cell pairs, if so, screening the similarity of the types of the plurality of pairs of matched cell pairs to screen out the matched cell pairs which are all one-to-one; and
the number of matched cell pairs in the selected matched cell pair group is used as the number of the repeated cells.
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| TW108108087A TWI704335B (en) | 2018-12-07 | 2019-03-11 | Automatic cell analysis device and operation method thereof |
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| JP2000501184A (en) * | 1995-11-30 | 2000-02-02 | クロマビジョン メディカル システムズ，インコーポレイテッド | Method and apparatus for automatic image analysis of biological specimens |
| TWI614500B (en) * | 2016-11-21 | 2018-02-11 | 國立清華大學 | Image registering and stitching method and image detection system for cell detection chip |
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| US4535284A (en) * | 1981-07-10 | 1985-08-13 | Coulter Electronics, Inc. | High and low frequency analysis of osmotic stress of cells |
| CN1590982A (en) * | 2003-09-01 | 2005-03-09 | 亚诺法生技股份有限公司 | Automated Optical Detection System for Cells |
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| TW202022345A (en) | 2020-06-16 |
| TWI704335B (en) | 2020-09-11 |
| CN109709346A (en) | 2019-05-03 |
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