JPS60157050A - Method for measuring t cell percentage of lymphocyte - Google Patents

Abstract

PURPOSE:To lessen manpower for a measuring method and to speed up said method by separating the video signal obtd. by sensing the image of a specimen for every condition from the shape of the cell data inputted preliminarily by utilizing a video processing technique and counting said signal. CONSTITUTION:A dyed specimen 1 is read via an optical system 2 by a line scanning type solid-state image pickup element 3 and is digitally converted in a measuring circuit 4, by which the specimen is discriminated to T cells, the lymphocytes except the T cells and other cells from the shape, etc. of the cells included in the specimen 1. The T cells and the lymphocytes except the T cells are counted in a counter circuit 5 and after both are counted by, for example, 200 pieces, the valve of 1/2 the number of the T cells is outputted to a display circuit 6. A control part 8 matches the timings in the processing speed of the respective parts. The percentage measurement is thus accomplished by utilizing the video processing technique, by which the measurement at a higher speed with less manpower is made possible.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Application of the Invention] The present invention relates to a method for measuring the percentage of T cells in lymphocytes, and particularly to a testing method for determining the percentage of T cells in lymphocytes using image processing technology.

[Background of the invention]

The increase, decrease, and atypicality of lymphocytes, a type of white blood cell, have long been useful in diagnosing infectious diseases such as whooping cough, but in recent years, advances in lymphocyte analysis have led to the diagnosis of new diseases such as immunodeficiency diseases. Progress is being made in elucidating the issue.

Lymphocytes are classified into T cells, B cells, and NULL cells based on their functions. In a normal person, the proportion of these substances is stable, but when there is a disease, the proportion changes.

Since the test for measuring the percentage of T cells in lymphocytes is a new test, no automated test method has been proposed yet. A typical method of testing that has been carried out in the past is that a laboratory technician observes a specimen prepared by the E rosette method using a microscope and performs the test. Next, this inspection process will be explained.

(1) Lymphocyte extraction: PB to collected heparinized blood
Add S and C0NRAY-FICoL, perform centrifugation, and extract lymphocytes.

(ii) Lymphocyte count adjustment: Count the number of lymphocytes using a hemocytometer and adjust the number of lymphocytes in the sample.

(tit) Rosette formation: After adding sheep red blood cells, culture to allow T lymphocytes to form rosettes.

(1v) Application of slide glass: After drawing a slide glass Hesmear, it is dried and stained with safranin.

(V) Microscopic examination: 20 lymphocytes were detected by microscopic observation.
Count 0 cells and calculate the percentage of lymphoid T cells to which sheep red blood cells are attached.

However, with this method, a laboratory technician observes the microscope and
Since T cells, which are lymphocytes, are counted from within the field of view, the test requires time and effort.

Therefore, it is desirable to automate the inspection as an even simpler method.

Therefore, the present inventors measured the shape of cells contained in the specimen for microscopic examination from the video signal output from reading the specimen with a line scan type solid-state image sensor, and determined whether the cells were T cells or lymphocytes other than T cells. , proposed an apparatus for calculating the percentage of T cells by distinguishing them from other cells (Japanese Patent Application No. 57-36250,
(See the specifications of Japanese Patent Application No. 57-41454).

By the way, the specimen stained by the rosette method is examined under a microscope.
If you look closely, you will see that the objects are color-coded into the following three types. That is, (a) a colorless background, (b) pale yellow-brown sheep red blood cells and human red blood cells, and (c) red lymphocytes and granulocytes.

In order to accurately identify cells, the video signals output from reading the specimen with an imaging device such as the line scan type solid-state imaging device are classified into the three categories (a), (b), and (c). Of these, it is necessary to measure areas (b) and (c).

However, conventionally, there have been the following two problems.

(,) The number of cells contained in the sample is 200 lymphocytes, 15,000 sheep red blood cells, and several hundred other cells, so when measuring the area (b), a huge number of sheep red blood cells will be measured. There must be.

(b) Among the cells contained in the sample, T cells tend to aggregate, so in the method of measuring areas (b) and (c) at the same time,
Counting the assembled T cells becomes troublesome.

[Purpose of the invention]

An object of the present invention is to solve these conventional problems and provide a method for measuring lymphocyte T cell percentages that can automatically and more easily and accurately measure the percentage of lymphocyte T cells.

[Summary of the invention]

In order to achieve the above object, 1 the lymphocyte T cell percentage measuring method of the present invention is based on a video signal obtained by imaging a specimen with an imaging device, and a predetermined first cell data of cell data contained in the specimen.
After measuring the shape that is greater than or equal to the output of the cell data, extract data in a range that is at least I pixels larger than the range of the cell data, and measure the shape that is greater than or equal to the predetermined second output of the data, and It is characterized by its separation into T lymphocytes, non-T lymphocytes, and other cells based on these values.

[Embodiments of the invention]

FIG. 1 is a block diagram of a lymphocyte T cell percentage measuring device showing an embodiment of the present invention.

The measurement device includes an optical system 2 for reading the sample 1, an automatic feed mechanism 7 for the sample 1, an automatic focus mechanism 9 for the optical system 2, a line scan type solid-state image sensor 3, a measurement circuit 4, and a counter circuit 5-'
It is composed of an RTA cell percentage display circuit 6 and a control section 8.

Specimen 1 is prepared by the E-rosette method, applied in a line with a width of 1 to 2 mm, fixed, and then stained.

The specimen 1 is moved by an automatic feed mechanism 7 while adjusting the focal position of the optical system 2 by an automatic focus mechanism 9. The line scan type solid-state image sensor 3 reads the specimen 1 through the optical system 2 and outputs a video signal according to its brightness.

The measurement circuit 4 digitizes the analog signal output from the line scan type solid-state image sensor 3, and further calculates the shape of the cells contained in the sample 1 into horizontal and vertical lengths.

It is measured by area, concentration, etc., and differentiated into T## cells, lymphocytes other than T cells, and other cells. Counter circuit 5 counts T cells and lymphocytes other than T cells, and counts both at 200%
After counting, the value of 172 T cells is output to the T cell percentage display circuit 6 for display. The control unit 8 adjusts the timing of the processing speed of each unit.

In this way, in the measuring device shown in Fig. 1, the image sensor 3 reads the sample 1 and outputs the video signal, which corresponds to the areas (a), (b), and (c), which are divided into the three color categories. After classifying into each type using the output of Only when it is determined that this is the case, the area corresponding to (c) above and the (mouth) area around it are measured.

FIG. 2 is an explanatory diagram of a cell discrimination method showing an embodiment of the present invention.

In Figure 2, (a), (b), and (c) correspond to three types of color-coded "colorless background J, r pale yellowish brown sheep red blood cells and human red blood cells" and "red lymphocytes and granulocytes." (A) is a pixel. Moreover, the symbols j~ and i~ on the outside of the screen are symbols indicating the coordinate positions in the vertical direction and the horizontal direction, respectively.

Here, is and ie respectively indicate the horizontal start coordinate position and end coordinate position of the area (C), and js and je respectively indicate the vertical start coordinate position and end coordinate position of the area (C). This is the symbol that indicates.

First, the measurement is performed on the area (c). Those having the region (c) include lymphocytes, granulocytes, and their low impurity content, and among these, lymphocytes have a horizontal length.

It can be determined based on the measurement items of vertical, surface length, area, and total concentration. The morphological characteristics of each particle are as follows.

That is, the stained area of lymphocytes is 3-8 μrn
The concentration is relatively uniform in size. In contrast, in the stained area of granulocytes, the size of the nucleus is larger than that of lymphocytes.

In addition, there are many types of impurities ranging from very large to small impurities, but their shapes are irregular and the difference in horizontal and vertical lengths is large. Therefore, by measuring standard lymphocytes or inputting numerical values, input the shape condition range that is determined as a lymphocyte in advance, and set the horizontal length, vertical length, and area.

It is determined whether the area (c) is a lymphocyte or not based on values such as total concentration.

If it is determined to be a lymphocyte, the position of an appropriate pixel in the area (c), for example, the position of pixel -1'A of (ieje), is recorded. Note that the pixel A to be recorded may be at any position as long as it is within the area (c). Next, based on the rectangle whose diagonal is the line connecting the two points of (151J sL ('ie-J e), we calculate the area to the outside by the number of pixels corresponding to the diameter of a sheep red blood cell + 1 pixel, that is, ( Measure the area corresponding to b) + (c).

The shape of a sheep red blood cell is 3 to 4 μm, and if one pixel is 1 μm wide, the above width is 4 to 5 pixels.

If the measurement result includes pixel A recorded in Figure 2, by subtracting the value of the area (c) determined above from this value, the sheep adhering to the lymphocyte T cell can be determined. It can be treated as red blood cells.

FIG. 3 is a correspondence diagram of a microscopic image and cell data showing an example of the present invention.

Figure 3 (a-1) (b-1) is a schematic diagram of a microscope image, and Figure 3 (a-2) (b-2) shows three types of images depending on the size of the image signal output from the imaging device. FIG. 3 is a diagram of cell data divided into regions. (a-1) (a-2) is a combination of two T cells, (b-1) (b-2) is a combination of sheep red blood cells and non-T lymphocytes be.

The analog signal of the mirror image of (a-1) is digitized, and it has color-coded areas (a), (b), and (c).
Obtain the cell data of a-2). In this case, there will be one area (b) and two areas (c), but
First, since the area (c) is measured, it is counted as two pieces. Of course, the horizontal length and vertical length are two areas.

It is determined from the values such as total concentration that the region (c) is a lymphocyte. In this case, since there are many regions (mouth) corresponding to sheep red blood cells, the region (c) can be identified as T lymphocytes.

When the analog signal of the microscope image in (b-1) is digitized, the cell data in (b-2) is obtained. According to this, the area (c) and the area (b) occur near it. First, the region (c) is determined to be lymphocytes, and these are measured and counted.

Next, when measuring the areas (B) and (C) within the range of 5 pixels outside, it is found that the area C) does not surround pixel A located in the area (C), or the pixel A Since it is not in contact with the area (c) containing the area, it can be determined that it is separate from the area (c). Furthermore, when comparing the region (c) containing pixel A with the values of the horizontal length, vertical length 2 area, total concentration, etc. that were previously measured in the region (c), it can be seen that it is a lymphocyte other than T. .

Note that in the method of the present invention, the shape of sheep red blood cells in contact with lymphocytes can be determined, so that impurities are not mistakenly identified as sheep red blood cells.

To measure the T cell percentage, as described above, count Tf41 cells and lymphocytes other than T cells in the area (c), count 200 of both, and then count 1/2 of the number of T cells.
This is possible by outputting .

〔Effect of the invention〕

As explained above, according to the present invention, after measuring the shape of the cell data included in the specimen that is equal to or larger than the first output, data in a range that is at least one pixel larger than the range of the cell data is extracted; Since the shape of the output of 2 or more is measured and the value is used to separate and count T lymphocytes, non-T lymphocytes, and other cells, lymphocytes other than Tjll cells can be counted without being mistaken as T cells. Since only sheep red blood cells near lymphocytes are measured, there is no need to measure all T cells, allowing for rapid measurement.Furthermore,
Because the lymphocytes are measured first, the number of lymphocytes can be counted accurately.

[Brief explanation of the drawing]

Fig. 1 is a block diagram of a lymphocyte T cell percentage measuring device showing an embodiment of the present invention, Fig. 2 is an explanatory diagram of a cell discrimination method showing an embodiment of the present invention, and Fig. 3 is an embodiment of the present invention. FIG. 2 is a correspondence diagram between a microscopic image and cell data. l: Sample, 2: Optical system, 3: Solid-state imaging device, 4: Measurement circuit, 5: Counter circuit, 6: T cell percentage display circuit, 7
: automatic feed mechanism, 9: automatic focus mechanism, 8: control section, (a): background area, (b): area of Higgi's red blood cells, (c)
: Area of lymphocytes and granulocytes. Figure 1 Figure 2 ■ 8 3

Claims (1)

[Claims] From a video signal obtained by imaging a specimen with an imaging device. After measuring the shape of the cell data included in the sample that is equal to or larger than the predetermined first output, extract the data in a range that is at least one pixel larger than the range of the cell data, and A method for measuring lymphocyte T cell percentage, which comprises measuring a shape greater than the output, and separating the cells into T lymphocytes, non-T lymphocytes, and other cells based on the above two measured values.