JPH08145988A - Method for identifying lymphocyte of different type - Google Patents

Abstract

PURPOSE: To quickly perform identification by automatically identifying an identification process along the step for each form feature of lymphocyte in different type using a computer. CONSTITUTION: The presence or absence of cellular nucleus in a cell is identified, a cell where the cellular nucleus exists is left, and a cell where no cellular nucleus exists is excluded. Then, it is judged whether the remaining cellular nucleus is lobate or not, the cell which is not lobate is left, and the cell which is lobate is excluded. Then, it is identified whether azure granule is present or absence in the remaining cell, the cell where no azure granule exists is left, and the cell where the azure granule exists is excluded. Then, the ratio of the remaining cell to basophil is identified, a cell with a high ratio is left, and a cell with a low ratio is excluded. Then, it is identified whether the cytoplasm of the remaining cytoplasm increases or not, the increasing cell is left, and the cell which does not increase is excluded. Then, the structure of chromatin of the remaining cell is identified, the cell in leptochromatic structure is left, and a cell of the chromatin in pachychromatic structure is excluded.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an atypical lymphocyte identification method capable of easily identifying atypical lymphocytes in blood in a short time. More specifically, the present invention provides a method for identifying atypical lymphocytes that can be suitably used for diagnostic indexes in the psychiatric field, psychiatric fields and immunological studies, grasping immunological pathological conditions after drug administration, toxicity tests, and the like. It is about.

[0002]

2. Description of the Related Art The discovery of atypical lymphocytes present in the peripheral blood of patients with schizophrenia has attracted attention as an immunological abnormality associated with an abnormality of the central nervous system. This atypical lymphocyte is found in all types of schizophrenic patients in various living environments and is morphologically similar to virus-infected cells, and is about half of the peripheral lymphocyte fraction of schizophrenic patients. It occupies one tenth. In addition, according to a double-blind test by a blood morphologist, about 95% of cases of schizophrenia diagnosed by a psychiatrist show the presence of atypical lymphocytes. Therefore, the identification of atypical lymphocytes can be used as an auxiliary diagnostic index in the diagnosis of schizophrenia.

However, conventionally, the identification process of this variant lymphocyte requires a considerable amount of time in the identification process, and since the morphology of blood cells must be identified, it also requires the cooperation of a hematological morphologist and is extremely specialized in identification. Knowledge and long-term experience were needed. This made it very difficult for a neuropsychologist to identify. In addition, little research has been conducted among psychoneurologists (psychiatrists) because the research content is related to hematology (blood morphology) and is too far from the field of psychiatry.

Therefore, the present invention was devised in order to solve the above-mentioned drawbacks of the prior art. It is possible to easily identify atypical lymphocytes in blood in a short time, and the department of neuropsychiatry. It is an object of the present invention to provide a new atypical lymphocyte identification method which can be suitably used as a diagnostic index for diseases such as schizophrenia and stress in a region.

[0005]

In order to solve the above-mentioned problems, the atypical lymphocyte identification method of the present invention is characterized by automatically identifying atypical lymphocytes according to the following 6 steps. Step 1: The presence or absence of cell nuclei is discriminated in the cells, the cells in which the cell nuclei exist are left, and the cells in which the cell nuclei do not exist are excluded.

Step 2: It is discriminated whether or not the cell nuclei of the cells left in step 1 are lobed, and the cells in which the cell nuclei are not lobed are left and the cells in which the cell nuclei are lobed are excluded. Step 3: The presence or absence of Azure granules is discriminated in the cells left in Step 2, cells without Azure granules are left, and cells with Azure granules are excluded. Step 4: Discriminate the cell nucleus to cytoplasmic ratio of the cells left in step 3, leaving cells with a high ratio and excluding cells with a low ratio.

Step 5: It is discriminated whether the cytoplasmic basophilicity of the cells left in step 4 is increased, and the cells with increased cytoplasmic basophilicity are left, and the cytoplasmic basophilicity of Exclude cells that have not increased. Step 6: Identify the structure of the chromatin of the cells left over in Step 5
Leaving cells that are chromatin of Leptochromatic structure, Pachyc
Exclude cells that are chromatin of the hromatic structure.

[0008]

The atypical lymphocytes in the blood can be easily identified in a short time by performing the identification process as described above by the computer step by step for each morphological feature of the atypical lymphocytes. More specifically, for example, in schizophrenia, P-cells as atypical lymphocytes and stress lymphocytes appear during stress. Therefore, this can be easily identified according to the above method. .

In the atypical lymphocyte identification method of the present invention, lymphocytes are extracted by the steps 1 to 3 as described above. Then, the atypical lymphocytes are re-identified with respect to the extracted lymphocytes. More specifically, in the lymphocyte re-identification step at this time, cells in which the cell nucleus is depressed and deformed and cells in which the cell nucleus is not depressed are identified. FIG. 1 illustrates a cell in which the nucleus identified here is depressed. In FIG. 1, (1) is an example of a normal cell, and (2) to (11) are examples of a cell having a depressed nucleus. The irregularity of the depressions is arranged from small to large.

Further, cells with and without eosinophilic granules are distinguished. FIG. 2 illustrates cells (12) to (17) in which the eosinophilic granules identified here are present. Note that FIG. 3 exemplifies cells having a high ratio and cells having a low ratio at the stage of distinguishing between cells having a high ratio of cell nucleus to cytoplasm and cells having a low ratio. In FIG. 3, (18) and (19) are normal cells with a low ratio, and (20) and (21) are cells with a high ratio.

FIG. 4 exemplifies strong basophilic cells at the stage of distinguishing between strongly basophilic cells having increased cytoplasmic basophilicity and cells having no increased basophilicity. In FIG. 4, (22) to (24) are normal cells, (25)
~ (33) are examples of strongly basophil cells. Further, in the present invention, cells and Pa that are chromatin of Leptochromatic structure are
It is distinguished from cells that are chromatin of chychromatic structure.

In practice, it is possible to carry out the above steps using a system consisting of a combination of a microscope, an image input device and a computer. That is, the atypical lymphocytes are determined from the test sample in which the procedures up to step 6 proceed. By identifying the presence of the atypical lymphocytes, it becomes possible to perform clinical determination, for example, determination of schizophrenia and the like, grasp of medical condition and pathological condition during treatment, and prescribing based on this.

[0013]

The present invention will be described in more detail with reference to the following examples. Of course, the present invention is not limited to the following examples. Venous blood smears were collected from 12 schizophrenic patients and 5 healthy volunteers. Fix these smears with methanol,

[0014]

Dyeing is performed, and P lymphocytes (P cells), which are a type of atypical lymphocytes, are identified by computer from these blood smears by the identification method of the present invention. Figure 5
FIG. 4 is a diagram showing a discrimination step when P lymphocytes (P cells), which are a type of atypical lymphocytes, are automatically discriminated by a computer. This will be described step by step. Step 1: Identify the presence or absence of cell nuclei in the cells. The cells with cell nuclei, WBC, are retained, and the cells without cell nuclei, erythrocytes and platelets, are excluded.

Step 2: Discriminates whether or not the cell nucleus of the cell left in Step 1 is divided, and determines that the cell is a cell having a band shaped nucleus which is a cell in which the cell nucleus is not divided.
Exclude Segmented Granulocytes, which are cells dividing the nucleus of the cell, leaving the monocytes and lymphocytes. Step 3: Special granules in the cells left in Step 2

[0017]

The presence or absence of azurophilic granules that are red-stained with a dye is identified, and cells that do not have azurophilic granules, monocytes and lymphocytes, are left, and granulocytes that have a bandshaped nucleus, which is a cell in which azurophilic granules are present, are excluded. Step 4: The ratio of cell nucleus to cytoplasm of the cells left in Step 3 is identified, and normal small lymphocytes and stimulated lymphocytes, which are cells with a high ratio, are left, and the ratio is low.
Exclude monocytes and normal medium and large sized lymphocytes, stress lymphocytes and NK lymphocytes.

Step 5: Stimulated lymphocytes, which are strongly basophil cells with increased cytoplasmic basophilicity, discriminating whether the cytoplasmic basophilicity of the cells left in step 4 is increased or not. , Normal small lymphocytes, which are cells with no increased cytoplasmic basophilicity. This left st
Downey type I cells and Downey typ for imulated lymphocytes
e III cells and P cells are included.

Step 6: The structure of the chromatin of the cells left in Step 5 was identified, and P cells and Downey type III cells having a structure rich in the leptochromatic structure, that is, heterochromatin, were left. , Downey type I cells having a structure rich in pachychromatic structure, that is, normal chromatin (euchromatin) are excluded. The above is the step of identifying P cells, and finally in step 6, P cells can be identified. Here, Downey type III cells have remained in step 6, but these Downey type III cells must have certain immunological complications such as influenza and autoimmune diseases, or have been infected with a virus. Since it does not exist, Downey type III cells can be excluded from the beginning by excluding those with such complications when collecting blood from schizophrenic patients. Therefore, P cells can be finally identified.

FIG. 6 is a photograph of an example of identified P cells. In FIG. 6, (1) is a normal lymphocyte of small size (10 μm or less), (2) is a normal lymphocyte of medium size (10 to 12 μm), (3) is a stress lymphocyte, and (4) is NK.
Lymphocytes, (5) to (15) are P cells, of which (5) to (9) are small P cells, (10) to (13) are medium P cells, and (14) and (15). Are large P cells.

FIG. 7 is an electron micrograph showing normal lymphocytes (A) and atypical lymphocytes (P-cells) (B). Symbols in FIG. 7B indicate the following. The length of the line segment is 1 μm. Ni: nucleolus M: mitochondria Ce: centriole Gr: granule From FIGS. 6 and 7, the morphological characteristics of P cells can be clearly seen by comparison with normal lymphocytes. That is, P
It can be seen that the cells have a high ratio of cell nucleus to cytoplasm, increased cytoplasmic basophilicity, and a chromatin having a Leptochromatic structure. Atypical lymphocytes can be easily identified in a short time by identifying such morphological characteristics stepwise by a computer.

Further, when the stress lymphocytes are further distinguished from the cells excluded in step 4 of the above-mentioned step, it can be distinguished by using the morphological characteristics of the stress lymphocytes shown in Table 1. FIG. 8 illustrates stress lymphocytes identified using the morphological features of Table 1. In FIG. 8, (34) to (35) are stress lymphocytes, and (36) the right end is an example of normal lymphocytes.

[0024]

[Table 1]

As an apparatus for the present invention, as shown in FIG. 9, for example, it may be constituted by an input device for observation by a microscope, a calculation device for discrimination and determination, a display device, an output device and the like. It is possible, and automatic identification is possible by the processing steps described in detail above. Figure 10
Shows an example of morphological analysis in automatic identification by a computer. It can be displayed by analyzing all of sizes (cells, nuclei), nuclear morphology, nuclear network structure, color tone of cytoplasm, special granules, etc. and calculating as numerical values.

[0026]

Since the present invention is configured as described in detail above, it has the following effects. Atypical lymphocytes can be easily identified in a short time by clearly identifying the identification process step by step for each morphological feature of the atypical lymphocytes and automatically identifying by a computer along the steps. Therefore, if this is used for schizophrenia diagnosis, even a psychiatrist can easily identify atypical lymphocytes in blood, and thus can be a useful diagnostic index for psychosis diagnosis.

[Brief description of drawings]

FIG. 1 is a photomicrograph replacing a drawing of an example of a cell in which the nucleus is depressed.

FIG. 2 is a photomicrograph instead of a drawing showing an example of cells in which eosinophilic granules are present.

FIG. 3 is a photomicrograph instead of a drawing showing an example of cells having a high ratio of cell nucleus to cytoplasm and a cell having a low ratio.

FIG. 4 is a microscope photograph instead of a drawing of a strongly basic cell example.

FIG. 5 is a diagram showing stepwise identification of P lymphocytes (P cells) by a computer according to an embodiment of the present invention.

FIG. 6 is a photomicrograph instead of a drawing of the example P cells identified in FIG.

FIG. 7 is a photomicrograph instead of a drawing showing normal lymphocytes and atypical lymphocytes (P-cells).

FIG. 8 is a micrograph, instead of a drawing, of an example of stress lymphocytes.

FIG. 9 is a schematic diagram showing a device configuration example for the method of the present invention.

FIG. 10 is a microscopic photograph as an alternative to a drawing showing an example of morphological analysis by a computer and a photograph including a halftone image displayed on a display.

Claims (1)

[Claims] 1. A method for identifying atypical lymphocytes, which comprises automatically identifying atypical lymphocytes according to the following 6 steps. Step 1: The presence or absence of cell nuclei is discriminated in the cells, the cells in which the cell nuclei exist are left, and the cells in which the cell nuclei do not exist are excluded. Step 2: Identify whether or not the cell nuclei of the cells left in Step 1 are lobed, and leave cells that are not lobed in the cell nuclei,
Exclude cells that have lobed cell nuclei. Step 3: The presence or absence of Azure granules is discriminated from the cells left in Step 2, cells without Azure granules are left, and cells with Azure granules are excluded. Step 4: Discriminate the cell nucleus to cytoplasmic ratio of the cells left in step 3, leaving cells with a high ratio and excluding cells with a low ratio. Step 5: It is discriminated whether or not the cytoplasm of the cells left in Step 4 has increased basophilicity, and the cells having increased cytoplasmic basophilicity are left, and the cytoplasmic basophilicity is increased. Exclude no cells. Step 6: The structure of the chromatin of the cells left in Step 5 is identified, and the cells that are the chromatin of the Leptochromatic structure are left and Pa
Exclude cells that are chromatin of the chychromatic structure.