Fluorescence image analysis device and fluorescence image analysis method

Abstract

The present invention provides a fluorescence image analysis technology that can improve the accuracy of determining whether a sample is positive or negative. A fluorescence image analysis device (1) is a fluorescence image analysis device that analyzes the fluorescence image of cells contained in a sample (10), and comprises: a light source (120-123) that irradiates light to the sample (10); a photographing unit (160) that photographs the fluorescence image of cells that emit fluorescence due to the irradiation of light; and a processing unit (11) that processes the photographed fluorescence image, wherein the processing unit (11) obtains the type of bright spot of fluorescence in the fluorescence image, and displays a plurality of positive types that are pre-corresponding to at least one of the measurement item and the labeling reagent on a display unit (13), and based on the obtained bright spot type and positive type, displays at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells, and the ratio of normal cells contained in the sample (10) on the display unit (13).

Description

Technical Field

[0001] The invention relates to a fluorescence image analysis device and a fluorescence image analysis method. Background Art

[0002] Patent document 1 discloses a method for processing cells when using a flow cytometer or the like for detection by fluorescence in situ hybridization (FISH). According to the FISH method, cells are stained by pretreatment in which a labeled probe is hybridized with a DNA sequence region of a detection target in the cell. Then, abnormal cells can be detected by detecting the fluorescence generated by the labeled probe.

[0003] Prior art literature

[0004] Patent Literature

[0005] Patent document 1: Japanese Patent Publication No. 2005-515408. Summary of the invention

[0006] Technical problem to be solved by the invention

[0007] In the above-mentioned abnormal cell analysis method, there are typical positive types and atypical positive types in the positive types. In addition, the atypical positive types sometimes also include other positive types caused by chromosomal abnormalities such as chromosome deletions. In order to accurately determine whether a sample is positive or negative for a specific disease, the operator must determine without omission which positive type of multiple positive types the cells contained in the sample match. Therefore, if a missed judgment occurs in the above-mentioned analysis method, the accuracy of determining whether the cell is abnormal may be reduced.

[0008] In view of the above, one of the objects of the present invention is to provide a fluorescent image analysis technology that can improve the accuracy of determining whether a sample is positive or negative.

[0009] Technical means to solve technical problems

[0010] Figure 1 , 3As shown in , 4 and 7, a fluorescence image analysis device (1) involved in a technical solution of the present invention is a fluorescence image analysis device for analyzing the fluorescence image of cells contained in a sample (10), which comprises: a light source (120-123) for irradiating light to the sample (10); a shooting unit (160) for shooting the fluorescence image of the cells emitting fluorescence due to the irradiation light; a processing unit (11) for processing the shot fluorescence image; and a display unit (13), wherein the processing unit (11) obtains the bright spot type of fluorescence in the fluorescence image, and displays a plurality of positive types pre-correlated with at least one of the measurement item and the labeling reagent on the display unit (13), and based on the obtained bright spot type and the plurality of positive types, at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells and the ratio of normal cells contained in the sample (10) is displayed on the display unit (13).

[0011] According to the above technical solution, the fluorescent image analysis device (1) obtains the type of fluorescent bright spot in the fluorescent image, and displays a plurality of positive types that are pre-corresponded to at least one of the measurement items and the labeling reagent on the display unit (13). Based on the obtained bright spot type and the plurality of positive types, at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells, and the ratio of normal cells contained in the sample (10) is displayed on the display unit (13). Therefore, it is possible to suppress the missed judgment of which positive type among the plurality of positive types the cells contained in the sample (10) correspond to. Therefore, it is possible to improve the accuracy of judging whether the sample (10) is positive or negative. In addition, the user can easily grasp at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells, and the ratio of normal cells contained in the sample (10).

[0012] like Figure 1 , 3 As shown in FIG. 4 , in the fluorescence image analysis device ( 1 ), the processing unit ( 11 ) displays all the multiple positive types that are preliminarily associated with at least one of the measurement item and the labeling reagent on the display unit ( 13 ) and enables the selection.

[0013] According to the above technical solution, the processing unit (11) displays all the multiple positive types that are pre-associated with at least one of the measurement items and the labeling reagent on the display unit (13) and enables selection. Therefore, the possibility of suppressing the missed determination of which of the multiple positive types the cells contained in the sample (10) correspond to is improved. Therefore, the accuracy of determining whether the sample (10) is positive or negative can be further improved.

[0014] like Figure 1 , 3As shown in and 4, in the fluorescence image analysis device (1), the processing unit (11) displays on the display unit (13) a plurality of positive types pre-corresponding to at least one of the measurement items and the labeling reagent and check boxes respectively configured to correspond to the positive types.

[0015] According to the above technical solution, the processing unit (11) displays a plurality of positive types pre-associated with at least one of the measurement item and the labeling reagent and check boxes respectively configured corresponding to the plurality of positive types on the display unit (13). Therefore, all the plurality of positive types pre-associated with at least one of the measurement item and the labeling reagent can be selected in the form of check boxes. Therefore, the operation of selecting the positive type by the operator becomes easy.

[0016] like Figure 4 As shown, it can be designed that in the above-mentioned fluorescent image analysis device (1), the processing unit (11) manages the authority of each user to operate a selection interface that can select multiple positive types, and displays the selection interface corresponding to the specific user who is granted the authority on the display unit (13).

[0017] According to the above technical solution, the processing unit (11) manages the authority of each user to operate the selection interface that can select a plurality of positive types, and displays the selection interface corresponding to the specific user who is granted the authority on the display unit (13). Therefore, the specific user who is granted the authority can operate the selection interface corresponding to the specific user. Therefore, it is possible to prevent other users from operating the selection interface and arbitrarily changing the positive type desired by the specific user.

[0018] like Figure 1 and 9 As shown, the fluorescent image analysis device (1) may further include a storage unit (12) for storing the determination results of the sample (10), and the processing unit (11) may display all the stored determination results on the display unit (13).

[0019] According to the above technical solution, the processing unit (11) displays all the stored judgment results on the display unit (13). This can improve the possibility of suppressing the missed judgment of which positive type the cells contained in the sample (10) correspond to among the multiple positive types. Therefore, the judgment accuracy of whether the sample (10) is positive or negative can be further improved.

[0020] like Fig. 9 and 10 As shown, it can be designed that in the above-mentioned fluorescence image analysis device (1), the processing unit (11) causes at least one information including the number of abnormal cells contained in the sample (10), the ratio of abnormal cells, the number of normal cells and the ratio of normal cells to be displayed together with the fluorescence image of the cells contained in the sample (10) on the display unit (13).

[0021] According to the above technical solution, the processing unit (11) displays at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells and the ratio of normal cells contained in the sample (10) together with the fluorescent image of the cells contained in the sample (10) on the display unit (13). Therefore, the user can confirm the fluorescent image of the cells contained in the sample (10) together with at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells and the ratio of normal cells contained in the sample (10).

[0022] like Fig. 9 and 10 As shown, it can be designed that in the above-mentioned fluorescence image analysis device (1), the processing unit (11) causes text information representing at least one of the proportion of abnormal cells and the proportion of normal cells contained in the sample (10) and a graphic image representing at least one of the proportion of the abnormal cells and the proportion of the normal cells to be displayed on the display unit (13).

[0023] According to the above technical solution, the processing unit (11) causes the display unit (13) to display text information indicating at least one of the ratio of abnormal cells and the ratio of normal cells contained in the sample (10) and a graphic image indicating at least one of the ratio of abnormal cells and the ratio of normal cells. Therefore, the user can check the text information indicating at least one of the ratio of abnormal cells and the ratio of normal cells contained in the sample (10) and the graphic image together.

[0024] like Fig. 9 and 10 As shown, it can be designed that in the above-mentioned fluorescent image analysis device (1), the processing unit (11) causes the relevant judgment result of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells contained in the sample to be displayed on the display unit (13) in a display method different from the relevant judgment results of other positive types.

[0025] According to the above technical solution, the processing unit (11) displays the relevant judgment result of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells contained in the sample on the display unit (13) in a display mode different from the relevant judgment results of other positive types. Therefore, the user can more easily determine the relevant judgment result of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells in the judgment result interface including various judgment results.

[0026] like Figure 7 As shown, in the above-mentioned fluorescence image analysis device (1), the plurality of bright spot types pre-associated with at least one of the measurement item and the labeling reagent can be designed based on the color and number of bright spots of fluorescence emitted from the fluorescent dye labeling the target site.

[0027] like Figure 1 , 3 As shown in , 4 and 7, a fluorescence image analysis method involved in a technical solution of the present invention is a fluorescence image analysis method for analyzing the fluorescence image of cells contained in a sample (10), which includes the following steps: a step of photographing a fluorescence image of cells that emit fluorescence due to irradiating light from a light source to the sample; a step of processing the photographed fluorescence image, wherein the processing step obtains the bright spot type of fluorescence in the fluorescence image, so that a plurality of positive types pre-corresponding to at least one of the measurement items and the labeling reagent are displayed on the display unit (13), and based on the obtained bright spot type and positive type, at least one information of the number of abnormal cells contained in the sample (10), the ratio of abnormal cells, the number of normal cells and the ratio of normal cells is displayed on the display unit (13).

[0028] According to the above technical solution, the processing step obtains the type of fluorescent bright spot in the fluorescent image, and displays a plurality of positive types that are pre-corresponded to at least one of the measurement items and the labeling reagent on the display unit (13). Based on the obtained bright spot type and positive type, at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells, and the ratio of normal cells contained in the sample (10) is displayed on the display unit (13). Therefore, it is possible to suppress the missed judgment of which positive type among the plurality of positive types the cells contained in the sample (10) correspond to. Therefore, it is possible to improve the accuracy of judging whether the sample (10) is positive or negative. In addition, the user can easily grasp at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells, and the ratio of normal cells contained in the sample (10).

[0029] like Figure 1 , 3 As shown in Figure 4, in the above-mentioned fluorescence image analysis method, the processing step can be designed to display all the multiple positive types pre-correlated with at least one of the measurement items and the labeled reagents on the display unit (13) and enable them to be selected.

[0030] According to the above technical solution, the processing step displays all the multiple positive types that are pre-associated with at least one of the measurement items and the labeling reagent on the display unit (13) and enables selection. Therefore, the possibility of suppressing the missed determination of which of the multiple positive types the cells contained in the sample (10) correspond to is improved. Therefore, the accuracy of determining whether the sample (10) is positive or negative can be further improved.

[0031] like Figure 1 , 3As shown in FIG. 4 , in the fluorescent image analysis method, the processing step may be configured to display on the display unit ( 13 ) a plurality of positive types pre-corresponding to at least one of the measurement item and the labeling reagent and check boxes respectively configured to correspond to the positive types.

[0032] According to the above technical solution, the processing step displays a plurality of positive types pre-corresponding to at least one of the measurement item and the labeling reagent and check boxes respectively configured corresponding to the plurality of positive types on the display unit (13). Therefore, all the plurality of positive types pre-corresponding to at least one of the measurement item and the labeling reagent can be selected in the form of check boxes. Therefore, the operator can easily operate the selection of the positive type.

[0033] like Figure 4 As shown, it can be designed that in the above-mentioned fluorescence image analysis method, the processing step manages the authority of each user to operate a selection interface that can select multiple positive types, and displays the selection interface corresponding to the specific user who is granted authority on the display unit (13).

[0034] According to the above technical solution, the processing step manages the authority of each user to operate the selection interface that can select multiple positive types, and displays the selection interface corresponding to the specific user who is granted the authority on the display unit (13). Therefore, the specific user who is granted the authority can operate the selection interface corresponding to the specific user. Therefore, it is possible to prevent other users from operating the selection interface and arbitrarily changing the positive type desired by the specific user.

[0035] like Figure 1 , 9 As shown in FIG. 11 , the fluorescent image analysis method may further include a step of storing the determination results of the sample ( 10 ), wherein the processing step displays all the stored determination results on the display unit ( 13 ).

[0036] According to the above technical solution, the processing step displays all the stored judgment results on the display unit (13). This can improve the possibility of suppressing the missed judgment of which positive type the cells contained in the sample (10) correspond to among the multiple positive types. Therefore, the judgment accuracy of whether the sample (10) is positive or negative can be further improved.

[0037] like Figures 9-11 As shown, it can be designed that in the above-mentioned fluorescence image analysis method, the processing step causes at least one information including the number of abnormal cells contained in the sample (10), the ratio of abnormal cells, the number of normal cells and the ratio of normal cells to be displayed on the display unit (13) together with the determined fluorescence image of the cells contained in the sample (10).

[0038] According to the above technical solution, the processing unit (11) displays at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells and the ratio of normal cells contained in the sample (10) together with the fluorescent image of the cells contained in the sample (10) on the display unit (131). Therefore, the user can confirm the fluorescent image of the cells contained in the sample (10) together with at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells and the ratio of normal cells contained in the sample (10).

[0039] like Figures 9-11 As shown, it can be designed as follows: in the above-mentioned fluorescence image analysis method, the processing step causes text information representing at least one of the proportion of abnormal cells and the proportion of normal cells contained in the sample (10) and a chart image representing at least one of the proportion of abnormal cells and the proportion of normal cells to be displayed on the display unit (13).

[0040] According to the above technical solution, the processing unit (11) causes the display unit (13) to display text information indicating at least one of the ratio of abnormal cells and the ratio of normal cells contained in the sample (10) and a graphic image indicating at least one of the ratio of abnormal cells and the ratio of normal cells. Therefore, the user can check the text information indicating at least one of the ratio of abnormal cells and the ratio of normal cells contained in the sample (10) and the graphic image together.

[0041] like Figures 9-11 As shown, in the above-mentioned fluorescence image analysis method, the processing step can be designed so that the relevant judgment result of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells contained in the sample is displayed on the display unit (13) in a display method different from the relevant judgment results of other positive types.

[0042] According to the above technical solution, the processing unit (11) displays the relevant judgment result of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells contained in the sample on the display unit (13) in a display mode different from the relevant judgment results of other positive types. Therefore, the user can more easily determine the relevant judgment result of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells in the judgment result interface including various judgment results.

[0043] like Figure 7 As shown, in the fluorescent image analysis method, the plurality of bright spot types to which at least one of the measurement item and the labeling reagent is preliminarily associated may be determined based on the color and number of bright spots of fluorescence emitted from the fluorescent dye labeling the target site.

[0044] Effects of the Invention

[0045] According to the present invention, the accuracy of determining whether a sample is positive or negative can be improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 is a perspective view of an example of the structure of a fluorescence image analysis device and a preprocessing device involved in an embodiment;

[0047] Figure 2 (a) is an exemplary diagram of the first to third images and a bright field image acquired by the fluorescence image analysis device according to the embodiment; Figure 2 (b) is an explanatory diagram of extraction of a nuclear region by the fluorescence image analysis device according to the embodiment; Figure 2 (c) and (d) are explanatory diagrams of extraction of a bright spot region by the fluorescence image analysis device according to the embodiment;

[0048] Figure 3 is a diagram showing an example of a functional block configuration of a processing unit of a fluorescence image analysis device according to an embodiment;

[0049] Figure 4 (a) is an explanatory diagram of an example of a specific interface for specifying measurement items and labeling reagents in a display unit of a fluorescence image analyzer according to an embodiment; Figure 4 (b) is an explanatory diagram of an example of a positive type selection interface on a display unit of the fluorescence image analysis device according to the embodiment;

[0050] Figure 5 (a) is an explanatory diagram of an example of a process of reading a code attached to a storage box storing a labeled reagent using a reading device according to an embodiment; Figure 5 (b) is an explanatory diagram of an example of a process of reading a code described in an attached document related to a labeled reagent using a reading device according to an embodiment;

[0051] Figure 6 is an explanatory diagram of an example of an input interface for inputting information related to bright spot types in a display unit of a fluorescent image analysis device according to an embodiment;

[0052] Figure 7 (a) to (d) are schematic diagrams showing examples of arrangement of bright spots of negative type, positive type 1, positive type 2, and positive type 3, respectively, according to the embodiment;

[0053] Figure 8 A and Figure 8 B is an exemplary diagram of target sites to which each probe targeting the BCR / ABL fusion gene is hybridized according to an embodiment;

[0054] Fig. 9 is an explanatory diagram of an example of a determination result interface in a display unit of a fluorescence image analysis device according to an embodiment;

[0055] Fig.10 It is magnified Fig. 9 A diagram showing a portion of the determination result interface shown;

[0056] Fig.11 is a flowchart for explaining an example of the operation of the processing unit of the fluorescence image analysis device according to the embodiment;

[0057] Fig.12 is a flowchart for explaining an example of a bright spot type acquisition process of a processing unit of the fluorescence image analysis device according to the embodiment;

[0058] Fig.13 is a flowchart for explaining an example of a bright spot type selection process of a processing unit of the fluorescence image analysis device according to the embodiment;

[0059] Fig.14 is a flowchart for explaining an example of determination processing by a processing unit of the fluorescence image analysis device according to the embodiment;

[0060] Fig.15 is a flowchart for explaining another example of determination processing by the processing unit of the fluorescence image analysis device according to the embodiment;

[0061] Fig.16 This is a flowchart for explaining an example of display control processing by the processing unit of the fluorescence image analysis device according to the embodiment. DETAILED DESCRIPTION

[0062] The preferred embodiments of the present invention are described below with reference to the accompanying drawings. In addition, the same reference numerals are assigned to the same elements, and repeated descriptions are omitted. In addition, unless otherwise specified, the positional relationships such as up, down, left, and right are based on the positional relationships shown in the accompanying drawings. In addition, the dimensional ratios of the accompanying drawings are not limited to the ratios shown in the drawings. In addition, the following embodiments are illustrative for illustrating the present invention, and the present invention is not limited to these embodiments.

[0063] The following embodiments are obtained by applying the present invention to an apparatus for measuring and analyzing a sample prepared in a pretreatment including a step of hybridizing a nucleic acid probe labeled with a fluorescent dye to a target site in a nucleic acid. Specifically, in the following embodiments, the target site in the nucleic acid is the BCR gene on chromosome 22 and the ABL gene on chromosome 9, and cells that have a translocation between chromosome 9 and chromosome 22 seen in chronic myeloid leukemia are detected as abnormal cells based on the FISH method. That is, in the following embodiments, for example, cells in which the BCR gene or the ABL gene is translocated to generate a BCR-ABL fusion gene are detected as abnormal cells.

[0064] Figure 1 The schematic structure of the fluorescence image analysis device 1 of this embodiment is shown. As an example, Figure 1 The fluorescent image analysis device 1 shown has a measuring device 100 and a processing device 200, and measures and analyzes a sample 10 prepared by pretreatment by a pretreatment device 300. The operator (user) centrifuges a blood sample collected from a subject using a cell separation medium such as Ficoll, and recovers the measurement target cells, i.e., nucleated cells. When recovering the nucleated cells, it is also possible to use a hemolytic agent to hemolyze red blood cells to leave nucleated cells instead of recovering the nucleated cells by centrifugation.

[0065] The pretreatment device 300 includes a mixing container for mixing a nucleated cell suspension obtained by centrifugation or the like with a reagent, a dispensing unit for dispensing the nucleated cell suspension and the reagent into the mixing container, a heating unit for heating the mixing container, etc. The pretreatment device 300 performs pretreatment to prepare the sample 10, and the pretreatment includes, for example, a process of labeling a target part in a cell collected from a subject with a fluorescent dye, and a process of staining the nucleus of the cell with a nuclear staining dye. Specifically, in the process of labeling the target part with a fluorescent dye, the target sequence is hybridized with a probe including a nucleic acid sequence having a complementary sequence to the target sequence and labeled with a fluorescent dye.

[0066] The FISH method uses more than one fluorescent dye to detect the target site on the chromosome. Preferably, the FISH method uses more than two fluorescent dyes to detect the target site on the first chromosome and the target site on the second chromosome (the "first" and "second" that modify "chromosome" do not refer to the chromosome number, but are a general concept of numbers). For example, the probe hybridized with the BCR locus is a nucleic acid having a sequence complementary to the base sequence of the BCR locus, which is labeled with a first fluorescent dye that produces a first fluorescence of a wavelength of λ21 due to irradiation with light of wavelength λ11. By using this probe, the BCR locus is labeled with the first fluorescent dye. The probe hybridized with the ABL locus is a nucleic acid having a sequence complementary to the base sequence of the ABL locus, which is labeled with a second fluorescent dye that produces a second fluorescence of a wavelength of λ22 due to irradiation with light of wavelength λ12. By using this probe, the ABL locus is labeled with a second fluorescent dye. The nucleus is stained with a nuclear staining dye that produces a third fluorescence of a wavelength of λ23 due to irradiation with light of wavelength λ13. Wavelengths λ11, λ12, and λ13 are so-called excitation light.

[0067] Specifically, as an example, the pretreatment device 300 includes the following treatments: a treatment for fixing cells so that the cells do not shrink due to water loss; a membrane permeation treatment for opening holes in the cells of a size that allows the probe to be introduced into the cells; a thermal denaturation treatment for heating the cells; a treatment for hybridizing the target site and the probe; a washing treatment for removing unnecessary probes from the cells; and a treatment for nuclear staining.

[0068] As an example, the measuring device 100 includes a flow cell 110, light sources 120 to 123, condensers 130 to 133, dichroic mirrors 140 to 141, a condenser 150, an optical unit 151, a condenser 152, and an imaging unit 160. The sample 10 flows in a flow channel 111 of the flow cell 110.

[0069] The light sources 120 to 123 irradiate the sample 10 flowing in the flow cell 110 with light. The light sources 120 to 123 are constituted by, for example, semiconductor laser light sources. Light having wavelengths λ11 to λ14 is emitted from the light sources 120 to 123, respectively.

[0070] The condensers 130 to 133 respectively condense the light of wavelengths λ11 to λ14 emitted from the light sources 120 to 123. The dichroic mirror 140 transmits the light of wavelength λ11 and refracts the light of wavelength λ12. The dichroic mirror 141 transmits the light of wavelengths λ11 and λ12 and refracts the light of wavelength λ13. In this way, the light of wavelengths λ11 to λ14 irradiates the sample 10 flowing in the flow path 111 of the flow cell 110. In addition, the number of semiconductor laser light sources possessed by the measuring device 100 is not limited to 1 or more. The number of semiconductor laser light sources can be selected from 1, 2, 3, 4, 5 or 6, for example.

[0071] When the sample 10 flowing in the flow cell 110 is irradiated with light of wavelengths λ11 to λ13, fluorescence is generated from the fluorescent dye that stains the cells. For example, when the first fluorescent dye that marks the BCR locus is irradiated with light of wavelength λ11, the first fluorescence of wavelength λ21 is generated from the first fluorescent dye. When the second fluorescent dye that marks the ABL locus is irradiated with light of wavelength λ12, the second fluorescence of wavelength λ22 is generated from the second fluorescent dye. When the nuclear dye that stains the nucleus is irradiated with light of wavelength λ13, the third fluorescence of wavelength λ23 is generated from the nuclear dye. When the sample 10 flowing in the flow cell 110 is irradiated with light of wavelength λ14, the light passes through the cells. The transmitted light of wavelength λ14 that passes through the cells is used to generate a bright field image. For example, in an embodiment, the first fluorescence is a wavelength band of green light, the second fluorescence is a wavelength band of red light, and the third fluorescence is a wavelength band of blue light.

[0072] The condenser 150 condenses the first to third fluorescent lights generated from the sample 10 flowing in the flow path 111 of the flow cell 110 and the transmitted light that has passed through the sample 10 flowing in the flow path 111 of the flow cell 110. The optical unit 151 has a structure in which four dichroic mirrors are combined. The four dichroic mirrors of the optical unit 151 reflect the first to third fluorescent lights and the transmitted light at slightly different angles from each other and separate them on the light receiving surface of the imaging unit 160. The condenser 152 condenses the first to third fluorescent lights and the transmitted light.

[0073] The photographing unit 160 is composed of a time delay integration (TDI) camera. The photographing unit 160 photographs the first to third fluorescences and the transmitted light, and outputs the fluorescence images corresponding to the first to third fluorescences and the bright field image corresponding to the transmitted light as photographing signals to the processing device 200. The fluorescence images corresponding to the first to third fluorescences are referred to as "the first image", "the second image" and "the third image" respectively below. The "first image", "the second image" and "the third image" are used to analyze the overlap of bright spots, so they are preferably the same size. The "first image", "the second image" and "the third image" can be color images or grayscale images.

[0074] The bright spot in this embodiment refers to a small fluorescent spot generated in a fluorescent image. More specifically, the bright spot refers to a fluorescent spot obtained from the fluorescent dye of the nucleic acid probe bound to the gene at the target site in the nucleus.

[0075] Figure 2 (a) is a diagram showing examples of first to third images and a bright field image acquired by the fluorescence image analysis device according to the embodiment. Figure 2 The black dot-shaped part in the first image of (a) represents the bright spot of the first fluorescence, that is, the target part marked by the first fluorescent dye. In the second image, although not as bright as the first image, dark gray dots are found in the light gray representing the nucleus. This represents the bright spot of the second fluorescence, that is, the target part marked by the second fluorescent dye. In the third image, the roughly circular nuclear area is represented in black. In the bright field image, the actual state of the cell can be observed. In addition, Figure 2 Each image in (a) is an example image obtained by placing pre-treated leukocytes on a slide and observing them under a microscope. In the fluorescent image, in the raw data, the higher the fluorescence intensity, the whiter the image appears, and the lower the fluorescence intensity, the darker the image appears. Figure 2The first to third images (a) are images obtained by inverting the levels of the captured raw data and expressing them in grayscale. As described above, when the sample 10 flowing in the flow chamber 110 is captured by the imaging unit 160, since cells flow in the flow channel 111 in a state of being separated from each other, a fluorescent image and a bright field image are acquired for each cell.

[0076] return Figure 1 As an example, the processing device 200 has a processing unit 11, a storage unit 12, a display unit 13, and an input unit 14 as a hardware structure. The processing unit 11 is composed of a processor (CPU). The storage unit 12 is composed of a memory (RAM) that can be read and written, a read-only memory (ROM) that stores computer programs and data, and a hard disk, etc. used in the operation area of ​​various processes of the processing unit 11. The processing unit 11 and the storage unit 12 can be composed of a general-purpose computer. The hard disk can be included in the computer or placed as an external device of the computer. The display unit 13 is composed of, for example, a display. The input unit 14 is composed of, for example, a mouse, a keyboard, a touch screen device, etc. The processing unit 11 transmits data to and from the storage unit 12 via a bus 15, and inputs and outputs data to and from the display unit 13, the input unit 14, and the measuring device 100 via an interface 16.

[0077] For example, the processing unit 11 reads out and executes various computer programs stored in the ROM or hard disk through the RAM, thereby processing fluorescent images of cells obtained by measuring the sample 10 of the measuring device 100 and controlling the operation of the display unit 13, the input unit 14, etc.

[0078] Figure 3 FIG. 1 is a diagram showing an example of a module structure of the processing unit 11 of the fluorescence image analysis device 1 according to the embodiment. Figure 3 As shown, the processing unit 11 functionally includes an acquisition unit 21 , a selection unit 23 , a determination unit 25 , and a display control unit 27 .

[0079] For example, the acquisition unit 21 acquires first to third images obtained by gray-reversing the raw data captured by the imaging unit 160 . The acquisition unit 21 stores the acquired first to third images in the storage unit 12 .

[0080] The acquisition unit 21 acquires the bright spot type of fluorescence in the fluorescence image. For example, it acquires the bright spot type of the first fluorescence in the first image based on the first fluorescence, and acquires the bright spot type of the second fluorescence in the second image based on the second fluorescence.

[0081] return Figure 2 , Figure 2 (b) is a diagram for explaining extraction of a nuclear region by the fluorescence image analysis device according to the embodiment. Figure 2(c) and (d) are explanatory diagrams of extraction of a bright spot region by the fluorescence image analysis device according to the embodiment; Figure 2 The third image shown on the left side of (b) and Figure 2 The first image shown on the left side of (c) and Figure 2 The second image shown on the left side of (d) is obtained from Figure 1 The same region of the sample 10 flowing through the flow cell 110 is shown.

[0082] like Figure 2 As shown on the left side of (b), when the third image is acquired, the acquisition unit 21 generates a Figure 2 (b) shows a graph of brightness and frequency. The frequency on the vertical axis represents the number of pixels. The acquisition unit 21 sets a brightness threshold in the graph. Then, Figure 2 As shown by the dotted line at the right end of (b), the acquisition unit 21 extracts the range where pixels with brightness greater than the threshold are distributed as the core area. In addition, in the third image, when two cores overlap, the first to third images related to the overlapping cells are not used to determine whether the preprocessing is appropriate or to determine abnormal cells, and are eliminated.

[0083] like Figure 2 As shown on the left side of (c), when the first image is acquired, the acquisition unit 21 generates a Figure 2 (c) shows a graph of brightness and frequency in the center. In this graph, the acquisition unit 21 sets the brightness threshold as the boundary between the bright spot and the background based on the Otsu method, for example. Then, Figure 2 As shown by the dotted line at the right end of (c), the acquisition unit 21 extracts the range of pixels with brightness greater than the threshold as the bright spot area. In addition, when extracting the bright spot area from the first image, the bright spot with an extremely small area, the bright spot with an extremely large area, and the bright spot not included in the first image are extracted. Figure 2 The bright spots in the region of the nucleus shown on the right side of (b) are removed.

[0084] like Figure 2 As shown on the left side of (d), when the second image is acquired, the situation is the same as the first image, as shown in FIG. Figure 2 As shown in the center of (d), the acquisition unit 21 creates a graph of brightness and frequency based on the brightness of each pixel on the second image. The acquisition unit 21 sets a brightness threshold in the graph and Figure 2 As shown by the dotted line on the right end of (d), the range of pixels with brightness greater than the threshold is extracted as the bright spot area. In addition, when extracting the bright spot area from the second image, the bright spot with an extremely small area, the bright spot with an extremely large area, and the bright spot not included in the image are extracted. Figure 2 The bright spots in the region of the nucleus shown on the right side of (b) are removed.

[0085] In addition, the acquisition unit 21 may not be Figure 2 (b) to (d) are not used to make a chart as shown in the center of Figures 1 to 3. Instead, the region of the nucleus is extracted from the third image by calculating the region of the nucleus according to the above steps, and the region of the bright spot is extracted from the bright spots of the first image and the second image. In addition, the extraction of the bright spot can be designed to determine the degree of integration between the distribution waveform of the normal bright spot and the region of the judgment object. If the degree of integration is high, the region of the judgment object is extracted as a bright spot. The acquisition unit 21 detects cells by extracting the region of the nucleus from the third image, but it can also detect cells based on the bright field image. When detecting cells based on the bright field image, the acquisition of the third image can also be omitted.

[0086] return Figure 3 , the selection unit 23 of the processing unit 11 selects at least one bright spot type from a plurality of bright spot types including one or more positive types that are pre-corresponded to at least one of the measurement items and the labeling reagents based on the instruction from the operator. An example of the bright spot type selection process is described below. For example, the processing unit 11 performs the following process: at least one bright spot type corresponding to the measurement items of the sample and at least one of the labeling reagents is selected from a plurality of bright spot types corresponding to the measurement items and at least one of the labeling reagents stored in the storage unit 12.

[0087] Figure 4 (a) is an explanatory diagram of an example of a specific interface SC1 for specifying measurement items and labeling reagents in the display unit 13 according to the embodiment. Figure 4 As shown in (a), in the specific interface SC1, for example, the item "measurement item" or the item "probe name" (labeling reagent) can be determined by a pull-down method. Regarding the "measurement item", for example, BCR / ABL fusion gene, ALK gene or chromosome 5 long arm deletion can be selected for each cell. Regarding the "probe name", for example, if the "measurement item" is BCR / ABL fusion gene, DF probe or ES probe can be selected. In addition, the details of BCR / ABL fusion gene, DF probe and ES probe will be described later.

[0088] The processing unit 11 displays a specific interface SC1 for determining the measurement items and the labeling reagent for the specific sample 10, that is, a specific interface SC1 that allows the operator to prioritize the content of the measurement items rather than the content of the labeling reagent. Figure 4As shown in (a), the processing unit 11 may also perform display control so that the item "measurement item" in the specific interface SC1 is placed above the item "probe name". In addition, the processing unit 11 may also highlight and display only the item "measurement item" in the specific interface SC1. In addition, the processing unit 11 may also perform control so that after the item "measurement item" is determined in the specific interface SC1, the item "probe name" corresponding to the content of the specific item "measurement item" is determined. In addition, it may also be designed that: in order to allow the operator to prioritize the content of the measurement item rather than the content of the labeled reagent, the processing unit 11 first displays the specific interface including the item "measurement item", and after the "measurement item" is determined, a specific interface including the item "probe name" different from the specific interface including the item "measurement item" is displayed again. At this time, the processing unit 11 may control so that when the specific interface including the item "probe name" is displayed, the specific interface including the "measurement item" is not displayed.

[0089] According to the above technical solution, the operator can prioritize the content of the measurement item rather than the content of the marking reagent. Therefore, after locking the bright spot type corresponding to the measurement item, the bright spot type corresponding to the marking reagent can be further determined. Therefore, the bright spot type can be selected efficiently.

[0090] Figure 4 (b) is an explanatory diagram of an example of the positive type selection interface SC3 in the display unit 13 according to the embodiment. Figure 4 As shown in (b), the processing unit 11 displays all one or more positive types that are pre-associated with at least one of the measurement item and the labeling reagent on the display unit 13. For example, the operator Figure 4 After confirming the measurement item "BCR-ABL" and the probe name "ES" in the specific screen SC1 shown in (a), Figure 1 The display unit 13 shown in FIG. Figure 4 (b) shows the positive type selection interface SC3. Here, Figure 1 Among the plurality of positive types stored in the storage unit 12 shown in FIG. 1 , the positive type corresponding to the measurement item “BCR-ABL” and the probe name “ES” is Figure 4 (b) shows the typical type G1R2F1, the minority type G1R1F2, the 9q deletion type G2R1F1, and the 9q・22q deletion type G1R1F1. Therefore, Figure 4(b) All the check boxes of each type are checked. That is, all the positive types showing the determination results are selected. However, for example, the operator can remove the check in the check box by operating the positive type selection interface SC3, thereby removing at least one positive type from the display object of the determination result. On the other hand, it can be designed that even if it is temporarily removed from the display object of the determination result, if the operator operates the positive type selection interface SC3 and presses the "display all" button BT, the check boxes corresponding to the "typical type G1R2F1", "minority type G1R1F2", "9q deletion type G2R1F1" and "9q・22q deletion type G1R1F1" are all checked. In addition, although not shown in the figure, the positive type selection interface SC3 can also have a button for removing all checks, such as a "display all" button.

[0091] in addition, Figure 4 The positive type selection interface SC3 shown in (b) only displays the positive type, but may also include the negative type corresponding to the measurement item "BCR-ABL" and the probe name "ES". That is, the processing unit 11 may display an interface including the bright spot type including the positive type and the negative type corresponding to the specific measurement item and the probe name.

[0092] The processing unit 11 can change at least one selected bright spot type based on an instruction from the operator. For example, the processing unit 11 can display the positive type selection interface SC3 on the display unit 13, so that the operator can change each positive type in a pull-down manner. Then, the operator can change at least one of the "typical type G1R2F1", "minority type G1R1F2", "9q deletion type G2R1F1" and "9q・22q deletion type G1R1F1" to another type by operating the positive type selection interface SC3.

[0093] According to the above technical solution, at least one selected bright spot type is changed based on the instruction from the operator. Therefore, at least one selected bright spot type can be freely changed.

[0094] The processing unit 11 manages the authority of each operator to operate the positive type selection interface SC3 that can select at least one positive type. For example, the processing unit 11 associates the identification information of the selection interface SC3 with the identification information of the operator and stores them in the storage unit 12. The processing unit 11 can display the positive type selection interface SC3 corresponding to the specific operator who has been granted the authority on the display unit 13 by referring to the information stored in the storage unit 12.

[0095] According to the above technical solution, a specific operator with authority can operate the positive type selection interface SC3 corresponding to the specific operator. Therefore, it is possible to prevent other operators from operating the positive type selection interface SC3 and arbitrarily changing the positive type desired by the specific operator.

[0096] Figure 5 (a) is an explanatory diagram of an example of a process of reading a code attached to a tank storing a labeled reagent using a reading device according to an embodiment. Figure 5 (b) is an explanatory diagram of an example of a reading process of a code recorded in an attached document related to a labeled reagent using a reading device according to an embodiment; Figure 5 As shown in (a) and (b), Figure 1 The fluorescent image analysis device 1 shown may also have a reading device R, which reads the code C attached to the storage box B storing the labeled reagent or the attached document D related to the labeled reagent. Then, Figure 3 The acquisition unit 21 of the processing unit 11 shown acquires information included in the code C read by the reader R. The information included in the code C includes, for example, information related to the probe name (labeling reagent) and at least one of the positive type and the negative type corresponding to the probe name.

[0097] In addition, the code C includes not only a one-dimensional code, that is, a barcode, but also a two-dimensional code, that is, a QR code (registered trademark), and may include other codes.

[0098] Figure 6 1 is an explanatory diagram of an example of an input interface RC1 for inputting information related to a bright spot type in the display unit 13 according to the embodiment. Figure 6 As shown in FIG. 1 , when the code C contains relevant information of a new bright spot type, the processing unit 11 displays an input interface RC1 for inputting relevant information of the new bright spot type on the display unit 13. Figure 5 As shown in (a) and (b), the reading device R reads the code C attached to the storage box B storing the labeling reagent or the document D related to the labeling reagent. Then, the processing unit 11 refers to the information pre-stored in the storage unit 12, such as the information related to the probe name (labeling reagent) and the information related to at least one of the positive type and the negative type corresponding to the probe name. When information other than the information pre-stored in the storage unit 12 is included in the code C, the processing unit 11 displays the input interface RC1 on the display unit 13 to allow the operator to input the relevant information of the new bright spot type.

[0099] exist Figure 6In the example, the relevant information of the input content (such as "measurement item", "probe name", "negative type name", "bright spot information" of "negative type name", "positive type name", and "bright spot information" of "positive type name") has been entered. However, when the input interface RC1 is displayed, it is usually in a state where no input is made. The operator refers to Figure 5 (b) The input contents recorded in the attached document D shown are entered (for example, manually input) for each item. Regarding the input contents, the attached document D of labeling reagents manufactured by other companies or sold on the market, for example, records the labeling position and the positive type and negative type when the labeling reagent is used. In addition, the code attached to the labeling reagent manufactured by our company (Sysmex Corporation) contains relevant information of the input contents. The information can be read by the reading device R, thereby automatically entering the relevant information of the input contents into the input interface RC1. In addition, the input contents of the input interface RC1 can be freely edited by the operator, and can also be managed as follows. Figure 4 The positive type selection interface SC3 shown in (b) can also only be edited by operators who have been given permission.

[0100] According to the above technical solution, the code (C) attached to the storage box (B) storing the marking reagent or the document (D) related to the marking reagent is read by the reading device (R). When the read code (C) contains relevant information of a new bright spot type, an input interface for inputting relevant information of the new bright spot type is displayed on the display unit (13). Therefore, relevant information of the new bright spot type can be obtained effectively and easily, and an input interface for inputting relevant information of the new bright spot type can be displayed.

[0101] It can be designed that the processing unit 11 displays the input interface RC1 on the display unit 13 in a predetermined display mode in order to urge the operator to enter the relevant information of the new bright spot type. For example, when the labeling reagent is manufactured by another company or is commercially available, as described above, the processing unit 11 allows the operator to manually input the relevant information of the input content. Thus, for example, "G" representing the first bright spot, "R" representing the second bright spot, and "F" representing the third bright spot, which are expected to be commonly entered at the "bright spot information" in the input interface RC1, can be pre-displayed. Specifically, when the input interface RC1 is displayed on the display unit 13, "G□R□F□" is pre-displayed at the "bright spot information" of the "positive type name", thereby urging the operator to enter 0, 1, 2, or 3 only in the "□ (space)" part.

[0102] According to the above technical solution, the burden on the operator when entering relevant information of a new bright spot type can be reduced.

[0103] return Figure 3The determination unit 25 of the processing unit 11 determines whether the acquired bright spot type corresponds to one or more positive types included in the selected bright spot type, based on the acquired bright spot type and the selected bright spot type.

[0104] An example of determination processing will be described below. As an example, a method of determining whether a cell is an abnormal cell having a chromosomal abnormality will be described.

[0105] Figure 7 (a) is an example of the arrangement of bright spots of normal cells without chromosomal abnormalities, that is, an example of bright spot types (negative types). Figure 7 (b) to (d) are examples of bright spot types (positive types) of abnormal cells. Figure 7 In any one of (a) to (d), each image is displayed in a state of being superimposed on the third image.

[0106] like Figure 7 As shown in (a), when there is no abnormality in chromosomes such as the translocation of the BCR locus and the ABL locus, each gene independently exists as a pair of alleles in one nucleus. Therefore, in the first image, there are two first bright spots in one nuclear region. In addition, in the second image, there are two second bright spots in one nuclear region. At this time, after the first image and the second image taken at the same size are overlapped and synthesized, in the synthesized image, the two first bright spots and the two second bright spots exist in one nuclear region without overlapping. Therefore, as shown in Figure 7 As shown in (a), cells with two first bright spots and two second bright spots in the nuclear region are judged to have no chromosomal abnormality, that is, normal cells with negative chromosomal abnormality.

[0107] An example of a positive type is described using a probe targeting the BCR / ABL fusion gene [Cytocell BCR / ABL Translocation, Extra Signal (ES) Probe (Sysmex Corporation)] (hereinafter sometimes simply referred to as "ES probe"). In addition, there are multiple types of probes targeting the BCR / ABL fusion gene.

[0108] Figure 8 A and Figure 8 B is a diagram showing an example of the target site to which each probe hybridizes. The BCR gene is located at chromosome 22q11.22-q11.23, and the probe hybridizing with the BCR locus is labeled with the first fluorescent light (e.g., green). The ABL gene is located at chromosome 9q34.11-q34.12, and the probe hybridizing with the ABL locus is labeled with the second fluorescent light (e.g., red). Figure 8 A represents the binding site of the above-mentioned ES probe, Figure 8B represents the binding site of Cytocell BCR / ABL Translocation, Dual Fusion (DF) Probe (Sysmex Corporation, Cat No. LPH007) (hereinafter sometimes simply referred to as "DF probe").

[0109] like Figure 7 As shown in (b), when part of the ABL locus moves to chromosome 9 due to translocation, in the first image, the first bright spot exists at two points in the nucleus, and in the second image, the second bright spot exists at three points in the nucleus. At this time, after the first image and the second image are synthesized, in the synthesized image, there is one first bright spot, two second bright spots, one first bright spot, and a bright spot of the fourth fluorescent light (for example, yellow) that overlaps with the second bright spot in one nucleus (fused bright spot). Therefore, Figure 7 The cells with bright spots as shown in (b) were judged to be abnormal cells that were positive for translocation of the BCR gene and the ABL gene, that is, chromosomal abnormality.

[0110] like Figure 7 As shown in (c), due to translocation, part of the BCR gene locus moves to chromosome 22, and part of the ABL gene moves to chromosome 9. In the first image, there are three first bright spots in the nucleus, and in the second image, there are three second bright spots in the nucleus. At this time, after synthesizing the first and second images, in the synthesized image, there is one first bright spot, one second bright spot, two first bright spots, and a fusion bright spot where the second bright spots overlap each other in one nucleus. Therefore, as Figure 7 The cells with bright spots as shown in (c) were judged to be abnormal cells with translocation between the BCR locus and the ABL locus, that is, positive for chromosomal abnormality.

[0111] like Figure 7 As shown in (d), when the ABL locus moves to chromosome 9 due to translocation, there are two first bright spots in the nucleus in the first image, and two second bright spots in the nucleus in the second image. At this time, after the first and second images are synthesized, in the synthesized image, there is one first bright spot, one second bright spot, and one first bright spot and a fused bright spot where the second bright spot overlap in one nucleus. Therefore, Figure 7 The cells with bright spots as shown in (d) were judged to be abnormal cells with translocation between the BCR locus and the ABL locus, that is, positive for chromosomal abnormality.

[0112] As described above, it is possible to determine whether each cell is an abnormal cell with chromosomal abnormality based on the position and number of each bright spot in the composite image that is a composite of the first image and the second image. Therefore, the processing unit 11 counts the number of positions of each bright spot in the composite image of each first image and the second image for each cell, that is, the number of first bright spots that do not overlap with the second bright spot, the number of second bright spots that do not overlap with the first bright spot, and the number of fused bright spots where the first bright spot and the second bright spot overlap with each other as the bright spot type of fluorescence in the fluorescence image. For example, in Figure 7 In (d), the bright spot type can be created with the number of single first bright spots being "1", the number of single second bright spots being "1", and the number of combined bright spots consisting of the first bright spots and the second bright spots being "1".

[0113] In addition, the first bright spot, the second bright spot, and the merged bright spot can also be displayed as colors. For example, the first bright spot of a single body can be displayed as green (G), the second bright spot of a single body can be displayed as red (R), and the merged bright spot can be displayed as yellow (F). The number of bright spots G, R, and F can be displayed immediately after G, R, and F, respectively, to make it a bright spot type. For example, in Figure 7 In (d), the highlight type can be displayed as "G1R1F1".

[0114] In addition, the number of the first bright spots can be the total number of the first bright spots in the first image, and the number of the second bright spots can be the total number of the second bright spots in the second image to create a fluorescent bright spot type for a fluorescent image. Figure 7 In (d), the number of the first bright spots in the first image is "2", the number of the second bright spots in the second image is "2", and the number of fused bright spots where the first bright spots and the second bright spots overlap with each other in the composite image is "1" to create the bright spot type, which is displayed as "G2R2F1" and has the same meaning.

[0115] In the composite image, it is possible to determine whether the first bright spot of the first image and the second bright spot of the second image overlap by determining the ratio of the area where the first bright spot and the second bright spot overlap each other, for example, by determining whether the ratio of pixels located at the same position (coordinate information (x, y)) as each pixel included in the second bright spot among the plurality of pixels included in the first bright spot is greater than a threshold. Alternatively, it is possible to determine whether the distance between the center point of the first bright spot (the position of the pixel with the highest fluorescence intensity) and the center point of the second bright spot (the position of the pixel with the highest fluorescence intensity) is less than a threshold.

[0116] In addition, the type of fluorescent bright spot in the fluorescent image acquired for each cell can be the number of colors for each bright spot in the synthesized image. That is, the color of each pixel of the first image is displayed with a green gradation (RGB value) based on the pixel value, and the color of each pixel of the second image is displayed with a red gradation (RGB value), instead of displaying each image in grayscale. Then, when the images are overlapped and synthesized, based on the combination of the RGB values ​​of each pixel of the synthesized image, when the cell is an abnormal cell, there is a green first bright spot, a red second bright spot, and a yellow fusion bright spot formed by the overlap of the first bright spot and the second bright spot in the nuclear region. Therefore, counting the number of colors for each bright spot as a bright spot type can also determine whether the cell is an abnormal cell.

[0117] As described above, the processing unit 11 determines whether a cell is an abnormal cell or a normal cell based on the bright spot type obtained for each cell. In the present embodiment, the storage unit 12 stores reference types (plural bright spot types including one or more positive types pre-associated with at least one of the measurement items and the labeling reagent) for determining whether a cell is an abnormal cell or a normal cell. The processing unit 11 compares the bright spot type obtained for each cell with the reference type (at least one bright spot type) selected by the selection unit 23 from the plurality of reference types stored in the storage unit 12, thereby determining whether each cell is an abnormal cell.

[0118] In addition, reference types include Figure 7 At least one of the bright spot type of fluorescence in the fluorescent image of abnormal cells with chromosomal abnormality (positive type) and the bright spot type of fluorescence in the fluorescent image of normal cells without chromosomal abnormality (negative type) is shown. In this embodiment, the reference type includes both the bright spot type of abnormal cells (positive type) and the bright spot type of normal cells (negative type).

[0119] Regarding the comparison processing of the bright spot type, when the bright spot type of the cell of the analysis object is consistent with the negative type, the processing unit 11 determines that the cell is a normal cell. On the other hand, when the bright spot type of the cell of the analysis object is inconsistent with the negative type, it is compared with the typical positive type. When it is consistent with the typical positive type, the cell is determined to be a typical abnormal cell. On the other hand, when it is inconsistent with the typical positive type, the cell is determined to be an atypical abnormal cell. The processing unit 11 repeatedly performs the same comparison processing on all the cells of the analysis object, and determines whether each cell is an abnormal cell or a normal cell. The processing unit 11 stores the determination results for each cell in the storage unit 12.

[0120] As described above, the fluorescence image analysis device 1 can be designed as follows: the bright spot type pre-corresponding to at least one of the measurement items and the labeled reagent also includes a negative type, and the processing unit 11 determines whether the sample 10 matches the negative type contained in the selected bright spot type based on the acquired bright spot type and the selected bright spot type.

[0121] According to the above technical solution, not only the positive type but also the negative type contained in the selected bright spot type can be referred to, so that the accuracy of determining the presence or absence of abnormality of cells contained in the sample can be further improved.

[0122] return Figure 3 The display control unit 27 of the processing unit 11 controls various interfaces to be displayed on the display unit 13. For example, various interfaces include the above Figure 4 (a) The specific interface SC1 shown, Figure 4 (b) The positive type selection interface SC3 shown, Figure 6 The input interface RC1 shown, described later Fig. 9 The judgment result interface RC3 is shown.

[0123] Fig. 9 1 is an explanatory diagram of an example of the determination result interface RC3 in the display unit 13 according to the embodiment. Fig. 9 As shown, the display control unit 27 displays the information related to the determination result for each analysis target cell on the display unit 13. As the information related to the determination result, for example, the processing unit 11 displays the fluorescent image (the composite image of the first image, the second image, and the third image) of the cell determined to be an abnormal cell and the fluorescent image (the composite image of the first image, the second image, and the third image) of the cell determined to be a normal cell on the display unit 13.

[0124] exist Fig. 9In the judgment result interface RC3, the fluorescence images of the cells detected based on a certain analysis method are displayed in a horizontal and vertical arrangement in the cells of the analysis object. The determination result interface RC3 displays the measurement items 33 used for the analysis of the cells, and is provided with an analysis method selection column 34 for selecting the analysis method. In the analysis method selection column 34, it is possible to select whether to detect the DF type as the analysis method for normal cells. In addition, in the analysis method selection column 34, it is possible to select whether to detect typical abnormal cells (ES majority type) or whether to detect atypical abnormal cells (ES minority type, ES deletion type), etc. as the analysis method for abnormal cells. In addition, the judgment result interface RC3 is provided with a display image selection column 38, which can select, for example, in a pull-down manner, an option to display the fluorescence images of cells determined as abnormal cells (positive), an option to display the fluorescence images of cells determined as normal cells (negative), and an option to display the fluorescence images of all analyzed cells for the cells detected based on the analysis method selected in the analysis method selection column 34.

[0125] For cells detected based on the analysis method selected in the analysis method selection column 34, the processing unit 11 displays the fluorescent image of the cell selected in the display image selection column 38 in the image display column 35 of the determination result interface RC3. In the fluorescent image of the cell displayed in the image display column 35, the Cell ID is displayed for each cell, and the determination result of whether it is an abnormal cell (positive) or a normal cell (negative) is also displayed. In addition, the fluorescent image of the cell determined to be a normal cell or the fluorescent images of all analyzed cells can be displayed according to the selection in the display image selection column 38.

[0126] Thus, the operator or the like can observe the fluorescent image of the cell determined as an abnormal cell through the display unit 13. In addition, when the cell determined as an abnormal cell is determined as a normal cell through the observation of the operator or the like, the processing unit 11 selects the normal cell from the fluorescent image of the abnormal cell displayed on the display unit 13 through the input unit 14 by the operator or the like, and corrects the abnormal cell determination result revised by the revision button ("Revise Judgement") 36 to a normal cell. Then, the processing unit 11 stores the revised determination result in the storage unit 12. Similarly, when the cell determined as a normal cell is determined as an abnormal cell through the observation of the observer such as the operator, the processing unit 11 selects the abnormal cell from the fluorescent image of the normal cell displayed on the display unit 13 through the input unit 14 by the operator or the like, and corrects the normal cell determination result revised by the revision button 36 to an abnormal cell. Then, the processing unit 11 stores the revised determination result in the storage unit 12. Thus, the detection accuracy of abnormal cells and normal cells can be improved. In addition, the processing unit 11 can also display the revised fluorescent image of the cell determined as an abnormal cell or a normal cell again on the display unit 13.

[0127] In this manner, the processing unit 11 displays, on the display unit 13 , a determination result screen RC3 including a determination result of whether the acquired bright spot type corresponds to one or more positive types included in the bright spot type.

[0128] According to the above technical solution, the operator can easily grasp the determination result.

[0129] The processing unit 11 generates information related to the determination result of the sample 10 based on the determination result of whether each cell is an abnormal cell. For example, the processing unit 11 performs processing to generate at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells, and the ratio of normal cells based on the analysis result of the analysis method selected in the analysis method selection column 34. For example, the ratio of the number of abnormal cells and the ratio of the number of normal cells may be a ratio relative to the number of all cells detected (the total value of the number of cells determined to be abnormal cells and the number of cells determined to be normal cells), or a ratio relative to the total number of analyzed cells.

[0130] The processing unit 11 stores at least one information of the number of abnormal cells, the ratio of abnormal cells, the number of normal cells, and the ratio of normal cells in the storage unit 12 and displays it on the display unit 13. Fig. 9 In an example of the determination result interface RC3, a determination result column 37 is provided, and the number and ratio of cells determined as abnormal cells and the number and ratio of cells determined as normal cells are displayed in the determination result column 37. Here, the number and ratio of typical abnormal cells (positive majority) whose determination result is "G2R3F1", the number and ratio of typical abnormal cells (positive minority) whose determination result is "G3R3F3", the number and ratio of atypical abnormal cells (positive atypical) whose determination result is "G2R2F1", and the number and ratio of normal cells (negative) whose determination result is "G2R2F0" are displayed in the determination result column 37. In addition, for example, after selecting an analysis method for detecting atypical abnormal cells (ES minority type, ES deletion type) in the analysis method selection column 34, the number and ratio of atypical abnormal cells (positive) whose determination result is the number and ratio of normal cells (negative) and the like can be displayed in the analysis result column 37.

[0131] like Fig. 9 As shown in the determination result column 37 , the processing unit 11 displays at least one information of the number of abnormal cells and the ratio of abnormal cells and the number of normal cells and the ratio of normal cells contained in the sample 10 on the display unit 13 .

[0132] According to the above technical solution, the operator can easily grasp at least one information of the number of abnormal cells and the ratio of abnormal cells and the number of normal cells and the ratio of normal cells contained in the sample 10 .

[0133] like Fig. 9 As shown in the image display column 35 and the judgment result column 37, the processing unit 11 displays at least one information including the number of abnormal cells contained in the sample 10, the ratio of abnormal cells, the number of normal cells and the ratio of normal cells together with the fluorescent image of the cells contained in the judged sample 10 on the display unit 13.

[0134] According to the above technical solution, the operator can confirm at least one information of the number and ratio of abnormal cells and the number and ratio of normal cells contained in the sample 10 together with the fluorescent image of the cells contained in the sample 10 .

[0135] like Fig. 9 As shown in the judgment result column 37, the processing unit 11 can display the relevant judgment results of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells (such as typical abnormal cells (positive majority)) on the display unit 13 in a display method different from the relevant judgment results of other positive types. For example, the relevant information of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells can be displayed in red, and the information of other bright spot types can be displayed in black. In addition, the relevant information of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells can be highlighted. In addition, the relevant information of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells can be displayed in bold.

[0136] According to the above technical solution, the operator can more easily determine the relevant judgment result of the positive type with the largest number of abnormal cells or the largest proportion of abnormal cells in the judgment result interface RC3 including various judgment results.

[0137] Furthermore, the processing unit 11 can generate various other information such as text information such as “possibly positive” or “possibly negative” as information related to the determination result of the sample 10 and display it on the display unit 13 .

[0138] Fig.10 It is magnified Fig. 9 FIG. 1 is a diagram showing a portion of the determination result interface RC3. Fig. 9 and Fig.10 As shown, the processing unit 11 can display the determination result interface RC including the chart display column 32 that displays the relevant information of the determination result in a chart on the display unit 13. Fig.10As shown, the "Type" related pie chart shows the ratio of typical abnormal cells, atypical abnormal cells and normal cells, the "Nega / Posi" related pie chart shows the ratio of abnormal cells to normal cells, and the "Positive" related pie chart shows the ratio of typical abnormal cells to atypical abnormal cells.

[0139] like Fig. 9 and 10 As shown, the processing unit 11 causes the display unit 13 to display text information indicating at least one of the ratio of abnormal cells and the ratio of normal cells contained in the sample 10 and a graph image indicating at least one of the ratio of abnormal cells and the ratio of normal cells. In addition, the display is not limited to a pie chart, and other graphs such as a bar chart may be used.

[0140] According to the above aspect, the operator can check the text information indicating at least one of the ratio of abnormal cells and the ratio of normal cells contained in the sample 10 together with the graph image. The operator can easily understand the ratio of abnormal cells or the ratio of normal cells by checking the graph image.

[0141] The following reference Fig.11 An example of a fluorescence image analysis method executed by the processing unit 11 based on a computer program defining a processing procedure for analyzing a fluorescence image of a cell will be described. Fig.11 is a flowchart for explaining an example of the operation of the processing unit of the fluorescence image analysis device according to the embodiment. Figure 1 The storage unit 12 is shown, but the program may be installed from a computer-readable removable recording medium (not shown) such as a CD-ROM, or downloaded and installed from an external server (not shown) via a network (not shown).

[0142] like Fig.11 As shown, for example, the processing unit 11 obtains the first to third images by inverting the hierarchy of the original data captured by the imaging unit 160 and displaying them in grayscale (step S1). The processing unit 11 obtains the bright spot type of fluorescence in the fluorescent image (step S3). The processing unit 11 displays one or more positive types that are pre-corresponded to at least one of the measurement item and the labeled reagent on the display unit 13 (step S5). The processing unit 11 displays the judgment result of the sample 10 on the display unit 13 based on the acquired bright spot type and positive type (step S7). Fig.12 For details of step S3, refer to Fig.13 For details of step S5, refer to Figure 14~Figure 16 The details of step S7 will be described.

[0143] Fig.12FIG. 1 is a flowchart for explaining an example of a bright spot type acquisition process performed by a processing unit of a fluorescence image analysis device according to an embodiment of the present invention. Fig.12 As shown, Figure 2 When the third image is acquired as shown on the left side of (b), Figure 3 The acquisition unit 21 shown in FIG. 1 is based on the brightness of each pixel on the third image, as shown in FIG. Figure 2 (b) is shown in the center, a graph of brightness and frequency is created (step S31). Figure 2 When the first image is acquired as shown on the left side of (c), the acquisition unit 21 obtains the first image based on the brightness of each pixel on the first image, as shown in FIG. Figure 2 (c) is shown in the center, a graph of brightness and frequency is created (step S33). Figure 2 When the second image is acquired as shown on the left side of (d), the acquisition unit 21 acquires the second image based on the brightness of each pixel on the second image, as in the case of the first image. Figure 2 As shown in the center of (d), a graph of brightness and frequency is created (step S35). The acquisition unit 21 extracts the core area from the third image and extracts the bright spot area from the bright spots of the first image and the second image (step S37). The acquisition unit 21 creates a bright spot type based on the extracted core area and bright spot area (step S39).

[0144] Fig.13 FIG. 1 is a flowchart for explaining an example of a bright spot type selection process performed by a processing unit of a fluorescence image analysis device according to an embodiment of the present invention. Fig.13 As shown, Figure 3 The display control unit 27 shown in FIG. 1 displays a specific screen SC1 (see FIG. 1 ) for specifying the measurement items and the labeling reagent on the display unit 13. Figure 4 (a) (step S51). The operator determines the measurement item in the specific interface SC1 displayed on the display unit 13 (step S53). After determining the measurement item, the display control unit 27 displays the specific interface SC1 on the display unit 13 so that the probe name corresponding to the content of the specific measurement item can be determined (step S55). The operator determines the probe name in the specific interface SC1 displayed on the display unit 13 (step S57). The selection unit 23 selects the positive type associated with the determined measurement item and probe name. The display unit 13 displays the positive type (step S59).

[0145] Fig.14 FIG. 4 is a flowchart for explaining an example of determination processing by a processing unit of a fluorescence image analysis device according to an embodiment of the present invention. Fig.14 As shown, Figure 3The selection unit 23 shown selects a reference type (at least one bright spot type) from a plurality of reference types stored in the storage unit 12 (step S71). In the determination unit 25, when the bright spot type of the cell to be analyzed coincides with the negative type, step S72 is yes, and the process proceeds to step S73 to determine that the cell is a normal cell. On the other hand, when the bright spot type of the cell to be analyzed does not coincide with the negative type, step S72 is no, and the process proceeds to step S74 to compare with the typical positive type. In the determination unit 25, when it coincides with the typical positive type, step S74 is yes, and the process proceeds to step S75 to determine that the cell is a typical abnormal cell. On the other hand, when it does not coincide with the typical positive type, step S74 is no, and the process proceeds to step S76 to determine that the cell is an atypical abnormal cell. The determination unit 25 repeatedly performs the same comparison process on all the cells to be analyzed, and determines whether each cell is an abnormal cell or a normal cell. The determination unit 25 stores the determination result for each cell in the storage unit 12.

[0146] Fig.15 This is a flowchart for explaining an example of determination processing by the processing unit of the fluorescence image analysis device according to the embodiment. Fig.15 The determination process shown is different from the determination process in that the bright spot type of the cell to be analyzed is determined to be consistent with the positive type. Fig.14 The determination process shown is different. Fig.15 As shown, Figure 3 The selection unit 23 shown selects a reference type (at least one bright spot type) from a plurality of reference types stored in the storage unit 12 (step S81). In the determination unit 25, regarding the comparison process of the bright spot type, when the bright spot type of the cell to be analyzed is consistent with the typical positive type, step S82 is yes, and the process proceeds to step S83 to determine that the cell is a typical abnormal cell. On the other hand, in the determination unit 25, when the bright spot type of the cell to be analyzed is inconsistent with the typical positive type, step S82 is no, and the process proceeds to step S84 to compare with the atypical positive type. In the determination unit 25, when it is consistent with the atypical positive type, step S84 is yes, and the process proceeds to step S86 to determine that the cell is an atypical abnormal cell. On the other hand, in the determination unit 25, when it is inconsistent with the atypical positive type, step S84 is no, and the process proceeds to step S85 to determine that the cell is a normal cell. The determination unit 25 repeatedly performs the same comparison process on all the cells to be analyzed, and determines whether each cell is an abnormal cell or a normal cell. The determination unit 25 stores the determination result for each cell in the storage unit 12 .

[0147] In addition, Fig.14 and Fig.15In the above determination process, there may be plural negative types. That is, in determining whether the bright spot type of the cell to be analyzed is consistent with the negative type, the bright spot type of the cell to be analyzed may be compared with plural different negative types. Fig.14 and Fig.15 In the embodiment, when the bright spot type of the cell of the analysis object does not match the positive type of the reference object or the bright spot type of the cell of the analysis object does not match the negative type of the reference object, the display unit 13 displays a message indicating that an error has occurred. Fig.14 and Fig.15 When the bright spot type of the cell of the analysis object does not match the positive type of the reference object or the bright spot type of the cell of the analysis object does not match the negative type of the reference object, other positive types or other negative types that are not the original reference object are read from the storage unit 12 and compared with the bright spot type of the cell of the analysis object.

[0148] Fig.16 FIG. 1 is a flowchart for explaining an example of display control processing of a processing unit of a fluorescence image analysis device according to an embodiment. Fig.16 As shown, Figure 3 The display control unit 27 shown in FIG. 1 acquires the determination result from the determination unit 25 or the storage unit 12 (step S91). The display control unit 27 generates display information for displaying the determination result on the display unit 13, and outputs the generated display information to the display unit 13 (step S93). The display unit 13 displays the display information based on the received display information. Fig. 9 The determination result interface RC3 is shown (step S95).

[0149] As described above, according to the above embodiment, the fluorescence image analysis device 1 acquires the bright spot type of fluorescence in the fluorescence image, displays a plurality of positive types that are pre-associated with at least one of the measurement item and the labeling reagent on the display unit 13, and displays the determination result of the sample 10 on the display unit 13 based on the acquired bright spot type and positive type. Therefore, it is possible to suppress missed determination of which positive type among the plurality of positive types the cells contained in the sample 10 correspond to. Therefore, it is possible to improve the accuracy of determination of whether the sample 10 is positive or negative.

[0150] <Other embodiments>

[0151] The above embodiments are used to facilitate understanding of the present invention and are not intended to limit the present invention. The present invention may be modified / improved (such as combining various embodiments or omitting some technical solutions of various embodiments) without departing from its main purpose, and the present invention also includes its equivalents.

[0152] Number Description

[0153] 1: fluorescent image analysis device, 10: sample, 11: processing unit, 12: storage unit, 13: display unit, 120-123: light source, 160: imaging unit, B: storage box, C: encoding, D: attached document, R: reading device.

Claims

1. A fluorescence image analysis device for analyzing the fluorescence image of cells contained in a sample, characterized in that have: a light source, irradiating light to the sample; an imaging unit that captures a fluorescent image of the cell that emits fluorescence due to being irradiated with the light; A processing unit, for processing the captured fluorescence image; Display unit; Wherein, the processing unit performs the following processing: Acquire the bright spot type of the fluorescence in the fluorescence image, Displaying a list of a plurality of positive types that are preliminarily associated with at least one of the measurement item and the labeling reagent on the display unit, accepts a selection from the list of plural positive types to be used in parsing, Based on the acquired bright spot type and the selected plurality of positive types, at least one information of the number of abnormal cells contained in the sample, the ratio of the number of abnormal cells, the number of normal cells, and the ratio of normal cells is displayed on the display unit.

2. The fluorescence image analysis device according to claim 1, characterized in that: The list displays all of the plurality of positive types that are preliminarily associated with at least one of the measurement item and the labeling reagent and enables selection.

3. The fluorescence image analysis device according to claim 2, characterized in that: The processing unit displays, on the display unit, the plurality of positive types that are preliminarily associated with at least one of the measurement item and the labeling reagent and check boxes that are arranged to respectively correspond to the plurality of positive types.

4. The fluorescence image analysis device according to any one of claims 1 to 3, characterized in that: The processing unit performs the following processing: For each user, management is performed to enable the user to select the plurality of positive types of selection interfaces. The selection interface corresponding to the specific user to whom the authority is granted is displayed on the display unit.

5. The fluorescence image analysis device according to any one of claims 1 to 3, characterized in that: It also includes a storage unit for storing the determination results of each cell included in the sample; The processing unit displays all the stored determination results on the display unit.

6. The fluorescence image analysis device according to any one of claims 1 to 3, characterized in that: The processing unit displays at least one of the number of abnormal cells and the ratio of abnormal cells and the number of normal cells and the ratio of normal cells contained in the sample on the display unit together with the fluorescent image of the cells contained in the sample.

7. The fluorescence image analysis device according to any one of claims 1 to 3, characterized in that: The processing unit displays text information indicating at least one of the ratio of the abnormal cells and the ratio of the normal cells contained in the sample together with a graph image indicating at least one of the ratio of the abnormal cells and the ratio of the normal cells on the display unit.

8. The fluorescence image analysis device according to any one of claims 1 to 3, characterized in that: The processing unit displays on the display unit a correlation determination result of a positive type having the largest number or ratio of the abnormal cells contained in the sample in a display mode different from that of correlation determination results of other positive types.

9. The fluorescence image analysis device according to any one of claims 1 to 3, characterized in that: The plurality of positive types preliminarily associated with at least one of the measurement item and the labeling reagent are determined based on the color and number of bright spots of fluorescence emitted from the fluorescent dye labeling the target site.

10. The fluorescence image analysis device according to any one of claims 1 to 3, characterized in that: The plurality of positive types preliminarily associated with at least one of the measurement item and the labeling reagent include typical positive types and atypical positive types.

11. A fluorescence image analysis method, which analyzes the fluorescence image of cells contained in a sample, characterized in that The steps include: a step of capturing a fluorescent image of the cell that emits fluorescence due to irradiation of light from a light source to the sample; A step of processing the captured fluorescence image; The processing steps are as follows: Acquire the bright spot type of the fluorescence in the fluorescence image, Displaying a list of a plurality of positive types that are preliminarily associated with at least one of the measurement item and the labeling reagent on the display unit, accepts a selection from the list of plural positive types to be used in parsing, Based on the acquired bright spot type and the selected plurality of positive types, at least one of the number of abnormal cells, the ratio of the number of abnormal cells, the number of normal cells, and the ratio of normal cells contained in the sample is displayed on the display unit.

12. The fluorescence image analysis method according to claim 11, characterized in that: The processing step displays all of the plurality of positive types that are preliminarily associated with at least one of the measurement item and the labeling reagent on the display unit and enables selection.

13. The fluorescence image analysis method according to claim 12, characterized in that: The processing step includes displaying, on the display unit, the plurality of positive types that are preliminarily associated with at least one of the measurement item and the labeling reagent and check boxes that are arranged to correspond to the plurality of positive types, respectively.

14. The fluorescence image analysis method according to any one of claims 11 to 13, characterized in that: The steps of the processing are as follows: For each user, management is performed to enable the user to select the plurality of positive types of selection interfaces. The selection interface corresponding to the specific user to whom the authority is granted is displayed on the display unit.

15. The fluorescence image analysis method according to any one of claims 11 to 13, characterized in that: The method further comprises the step of storing the determination results for each cell contained in the sample. In the processing step, all of the stored determination results are displayed on the display unit.

16. The fluorescence image analysis method according to any one of claims 11 to 13, characterized in that: The processing step causes at least one of the number of abnormal cells and the ratio of abnormal cells, the number of normal cells and the ratio of normal cells contained in the sample to be displayed on the display unit together with the fluorescent image of the cells contained in the sample.

17. The fluorescence image analysis method according to any one of claims 11 to 13, characterized in that: The processing step causes the display unit to display text information indicating at least one of the ratio of the abnormal cells and the ratio of the normal cells contained in the sample together with a graph image indicating at least one of the ratio of the abnormal cells and the ratio of the normal cells.

18. The fluorescence image analysis method according to any one of claims 11 to 13, characterized in that: The processing step causes the display unit to display the correlation determination result of the positive type having the largest number or the largest proportion of the abnormal cells contained in the sample in a display mode different from that of the correlation determination results of other positive types.

19. The fluorescence image analysis method according to any one of claims 11 to 13, characterized in that: The plurality of positive types preliminarily associated with at least one of the measurement item and the labeling reagent are determined based on the color and number of bright spots of fluorescence emitted from the fluorescent dye labeling the target site.

20. The fluorescence image analysis method according to any one of claims 11 to 13, characterized in that: The plurality of positive types preliminarily associated with at least one of the measurement item and the labeling reagent include typical positive types and atypical positive types.

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