JPS5842899B2 - Hatsuketsukiyuuno Tankyuurinpaiyubenbetsouchi - Google Patents

Description

[Detailed Description of the Invention] There are three types of white blood cells: basophils, eosinophils, neutrophils, monocytes, and lymphocytes, each of which is produced in different parts of the body and has different functions. Ru.

Therefore, by counting the white blood cells in the blood by type, or by understanding the changes in each type of white blood cell,
It can contribute to the diagnosis of diseases.

According to the method for classifying and detecting leukocytes according to the present invention, basophils, eosinophils, and neutrophils are first separated, and finally monocytes and lymphocytes are left.

Therefore, this invention, as part of this classification detection method,
The aim is to realize a device that distinguishes between monocytes and lymphocytes.

The present invention will be explained below based on the illustrated embodiments.

In FIG. 1 L, 1 is a sample consisting of a preparation containing blood cells, which is placed on a stage 2 that is movable in the X direction, and passed through a hole 3 made in the stage 2. It is illuminated by a light source 4 through a reflecting mirror 5 and a condensing lens 6.

A projection microscope 7 is arranged above the sample 1, and this microscope 7 includes an objective lens 8, an eyepiece lens 9, a reflector 10,
It has a screen 11, and an enlarged image of the sample 1 is displayed on the screen 1.
Project on 1.

On the back side of the screen 11, as shown in FIG. 2, a large number of light guides 13a to 13n are arranged in a row in the Y direction perpendicular to the moving direction X of the sample image 12 as the mounting table 2 moves. It is combined with L.

The other ends of these light guides are respectively light receiving elements 14a to 14.
The light receiving signals of each element are supplied to AND circuits 15a to 15n, respectively.

Reference numeral 16 denotes a time division circuit, which generates time division viruses a - n at different times in sequence in synchronization with the clock pulse given as an input 17, and outputs them to AND circuits 15a to 15a, respectively.
15n to time-divisionally sample the light-receiving signals of each of the light-receiving elements 14a to 14n.

The sample outputs of these AND circuits 15a to 15n are
are combined into a common output line 18 into a series of sample scanning signals S.

For example, the light guide 13a is located at the position of the chain line 19 in FIG.
When the tip of ~13n is present, the sample scanning signal S in the figure is obtained by time-divisional sampling of the light receiving element.

Each pulse in the sample scanning signal S transmits the transmitted light beam of each part of the white blood cell along the chain line 19 to each of the light receiving elements 14a to 14, respectively.
14n.

Further, the time division circuit 16 supplies the X direction drive pulse X to the X direction drive mechanism 20 at every blank period when the generation of the time division signals a to n has completed, and moves the sample stage 2 in the X direction. Move it a small distance.

FIG. 3 shows the overall configuration of a leukocyte classification and detection system in which the present invention is implemented. From the sample scanning signal S obtained by the sample scanning device 21 shown in FIG. Globules and neutrophils are classified and removed.

Although the basophil detector does not have a nucleus, it does have a large number of granules, so if an amplitude level higher than the envelope of the sample scanning signal S is taken out and given to the counter, other A significantly higher count is shown compared to seed leukocytes.

Based on this principle, the basophil detector 22 detects basophils and detects whether the white blood cells being detected are basophils (and sometimes emit a signal Qb indicating that they are not basophils). Occur.

Eosinophils have a much lower light transmittance in their cell parts outside the plate than other types of leukocytes, so the sample scan signal S has reached the level where cell parts of other types of leukocytes can also be detected. When the number of pulses and the number of pulses reaching the second level, where only nuclei or granules are detected in other types of leukocytes, are counted separately, both counts are almost equal for eosinophils, whereas for other types of leukocytes, It will appear very different.

The eosinophil detector 23 uses this principle to detect eosinophils, and sometimes the white blood cells being detected are eosinophils.
A signal Qe indicating this is generated.

Also, when comparing neutrophils with monocytes and lymphocytes, monocytes and lymphocytes have relatively simple nuclei, whereas neutrophils have relatively simple nuclei.
Neutrophils have complex-shaped nuclei.

This means that neutrophils have a large nuclear circumference relative to their nuclear area, and according to the inventor's research, when N is the magnification used for measurement, it is shown in Figure 4. 1 5, the ratio of nuclear circumference to nuclear area is 2.13←−−0cm−
With 1600 as the boundary, monocytes and lymphocytes are distributed in the low to low range, and neutrophils are distributed in the high range.

The neutrophil detector 24 calculates the nuclear circumference and nuclear area based on the sample scanning signal S, derives the ratio -1, compares this with the above constant 2.13 (-), 600 Signal Qb indicating that the above ratio is more dog than the above constant and that it is not a basophil, and signal σ7 indicating that it is not an eosinophil.
When the detected white blood cell is a neutrophil, it is determined that the detected white blood cell is a neutrophil, and when the detected white blood cell is not a neutrophil, a signal Qn indicating this is generated.

Therefore, the signal Qmll! obtained by processing each output of the detectors 22, 23, 240 by the AND circuit 25! This signal indicates that the white blood cells being detected are neither basophils, acidophils, nor neutrophils; in other words, they are either monocytes or lymphocytes.

Now, when comparing monocytes and lymphocytes, the nuclei of lymphocytes are often circular, square, oval, semicircular, etc., as shown in Figure 5 a, b, c, and d, and the nuclei of monocytes are As shown in Figure 5 e, f, and g, there are many L-shaped shapes such as heart-shaped, U-shaped, and linear.

Therefore, regarding the various types of nucleation shown in Figure 5, The following results were obtained.

Therefore, it was found that monocytes and lymphocytes can be distinguished by utilizing the following: lymphocytes: R2O, 204 monocytes: R≧0.207.

Here, the polar moment of inertia (J) around the center of gravity G is the sixth
In the figure, L is a small area at a distance r from the center of gravity G, a
The moment of inertia r2aA that A has around the center of gravity G is integrated over the entire area A of the white spot sphere nucleus. The method for determining the moment of inertia) will be explained below.

The sample scanning signal S illustrated in FIG. 2 is compared with a constant voltage Vr supplied by a constant voltage source 27 in a comparator 26 in FIG.
”, and when the voltage does not exceed Vr, the signal “0” is output.
” is output.

Therefore, if a nucleus as shown by the broken line 29 exists in the scanning field of view 28, the distribution of the signal “1” and tt Ojj will be the seventh
It will look like the illustration.

While the first and second stages are being scanned from above in the Y direction within the scanning field of view 28 in FIG. 7, the output signals of the comparator 26 are all 'O'; At the beginning, the upper end of the nucleus 29 is detected and a signal "1" appears in a portion.

The stage where the signal "1" first appears is set as Xl-=1, and the following stages are sequentially set as X1=2.3.4, . . . .

Then, for each stage, the number Eyi in which the signal "1" appears is counted.

Based on these values Xi and Fyi, [Xl
−0,5], (Fyi (X i −0,5) :l,
C(Xi-0,5)2), [Fyi(Xi-0,
5) Calculate 2:), Fyi, (Fyi (Xi-
0,5) ) and (Fyi (X i -0,5)2), the sum Σ of all stages is determined.

ΣFyi (-X i -0,5)2 obtained in this way
represents the moment of inertia around the upper end of the nucleus 29, that is, Xl-0,5.

If this is moved to the position of the center of gravity G using the parallel axis theorem, FIG. 8 shows an apparatus for calculating Iy based on the above-mentioned principle.

The output signal of the comparator 26 is applied to a flip-flop 31 which outputs a signal P1'' during scanning of the stages in the scanning field 28.
It is set when the time division circuit 16 appears, and is reset by the (n+1)th signal of the time division circuit 16 after completing the scanning of that stage.

Then, when the nth signal prior to reset appears, if it is in the set state, the AND circuit 32 produces an output,
It is added to the counter 33.

That is, the counter 33 calculates the number of stages in which the signal 1"1" appears, ie, Xi.

The output Xi of the counter 33 is introduced into a subtraction circuit 34, where the constant 0.5 given from the constant setter 35 is subtracted,
Produces a differential output (Xi-0,5).

This difference output is sent to the 2nd east circuit 36 and squared.

The output signal of the comparator 26 is also supplied to a counter 37 and counted.

This counter 37 is reset by the (n + 1)th signal of the time division circuit 16 every time the Y-direction scan is performed, so the count value is determined by the appearance of the signal "1" at each stage in the scanning field of view. The number of times, that is, Fyi.

The signal Fyi is multiplied by the output of the subtraction circuit 34 in a multiplier circuit 38, and the product is accumulated in a counter 39 to produce ΣFyi(X i -0,5).

Further, the signal Fyi is inputted to the second east circuit 36 in the multiplier circuit 40.
The product is accumulated by the counter 41 by the output of ΣFyi(Xi-0,5)2.

The signals supplied from the counter 42 and the east comparator 26 are accumulated over the entire scanning field of view to produce ΣFyi.

This is a count value corresponding to the area of the nucleus.

The output of the counter 39 is squared in a 2-east circuit 43 and divided by the output of the counter 42 in a division circuit 44 .

45 is a subtraction circuit, which divides the output of the counter 41 from the division circuit 4.
The high output of 4 is subtracted to produce a differential output Iy.

As is clear from the above calculation process, This difference output matches Equation (3).

After completing this calculation, counters 33, 39, 41, 4
2 is reset.

Referring again to FIG. 3, the output of comparator 26 is output to AND circuit 4.
6, the signal is supplied to an Iy arithmetic unit 47 shown in FIG.

During the period of the write signal applied from the terminal 50, the storage device 48 stores the same pattern as the scanning field of view 28 shown in FIG. During the signal period, the stored pattern is read by fast scanning in the X direction and slow scanning in the Y direction.

These write signals and read signals are both sent to the sample scanning device 21.
The write signal continues for one cycle of low-speed scanning in the X direction at , the write signal appears for the first week, and the read signal appears for the following week.

■The outputs of the y calculator 47 and the Ix calculator 49 are added together in the adder 52, and as shown in equation (2), a sum output corresponding to the polar moment of inertia J around the center of gravity of the nucleus is generated. Ru.

The output of the comparator 26 which has passed through the AND circuit 46 is also counted by a counter 53 to produce an output corresponding to the kernel area A.

Since the output ΣFyi of the counter 42 in the Ly calculator 47 is equal to A, this counter 4 is used instead of the counter 53.
You can use 2 L.

The output corresponding to the above ゑA is squared in the 2 east circuit 54, and the above-mentioned sum output J is divided in the divider 55, and (1
) produces a high output corresponding to R according to the equation.

The high output R is compared with a signal corresponding to the numerical value 0.206 supplied by the digitizer 57 to the comparator 56, so that R is 0.206.
When it is more dog than 206, turn to AND circuit 58,
Also, when R is smaller than 0.206, the AND circuit 59
Sends the comparison output to.

In addition to the above comparison output, the AND circuits 58 and 59 are supplied with the output Qml of the AND circuit 25 and the discrimination control signal applied to the terminal 60, and output the monocyte detection signal Qm and the lymphocyte detection signal Ql, respectively. Send as.

Note that the adder 52 and the counter 53 are reset by a reset signal, along with the counters 33, 39, and 4L42 in FIG.

In the above-described apparatus, the same white blood cells are repeatedly scanned twice by the sample scanning device 21.

When the scanned white blood cells are monocytes or lymphocytes, the basophil detector 22, eosinophil detector 23, and neutrophil detector 24 each output a signal Qb each time each scan is completed.

The AND circuit 25 generates the AND output Q
Generate ml.

During the first scan, a write signal is sent from the terminal 50 to the memory 48.
Since the pattern of the nucleus 29 as illustrated in FIG. 7 is stored in the memory 48, the terminal 5
Since the readout signal is not applied to L.1, the Ly calculator 47 and counter 53 do not operate.

During the second scan, the write signal at the terminal 50 disappears and the read signal is applied from the terminal 51, so the Lx calculator calculates the value of Lx based on the signal read from the memory 48. Based on the signal of the comparator 26 which has been calculated and passed through the AND circuit 46,
- The Ly calculator 47 calculates the value of Ly.

Then, the calculated Ly and Lx are sent to the adder 52 at °C.
produces a sum output J, and the sum output J is sent to the divider 55.
yields a high power R.

At the stage where the second scan is completed, if the high output R is smaller than 0.206, it is directed to the AND circuit 58, and again 0, 2
When it is smaller than 06, the output of the comparator 56 is sent to the AND circuit 59 and becomes L.

As mentioned above, R≧0.207 for monocytes and R2O,204 for lymphocytes, so if the scanned white blood cells are monocytes, the AND circuit 58 is used, and if the scanned white blood cells are lymphocytes, the AND circuit 59 is used. The output will be given.

At this point, the AND circuit 25 supplies a signal Qml indicating that the white blood cell is a monocyte or a lymphocyte to the AND circuits 58 and 59.

Therefore, when a discrimination control signal is applied to the terminal 60, if the scanned white blood cell is a monocyte, the AND circuit 58 outputs the monocyte detection signal Qm.
If it is a lymphocyte, the AND circuit 59 generates a lymphocyte detection signal Ql.

Once this discrimination is completed, a reset signal is supplied to reset the counters 33, 39, 41, 42, 53 and the adder, and the test is started for the next leukocyte in the sample.

As described above, by carrying out the present invention, it is possible to classify and detect monocytes and lymphocytes in cooperation with the means for classifying and detecting basophils, eosinophils, and neutrophils, and to detect all white blood cells. types of classification detection can be completed.

Therefore, it can greatly contribute to disease diagnosis by testing white blood cells.

[Brief explanation of the drawing]

The figures show an embodiment of the present invention; FIG. 1 is a configuration diagram of a sample scanning device, FIG. 2 is an explanatory diagram of a sample scanning mode and a sample scanning signal, and FIG. Figure 4 is a distribution curve diagram of monocytes, lymphocytes, and neutrophils, Figure 5 is a model diagram of various white blood cell nuclear patterns, and Figure 6 is the center of gravity G of white blood cell nuclei.
FIG. 7 is an explanatory diagram of the polar moment of inertia around -, FIG. 7 is an explanatory diagram of a quantized white blood cell nucleus pattern and calculation values based thereon, and FIG. 8 is a configuration diagram of a moment of inertia calculator. 12... White blood cells, 21... Sample scanning device, 22...
・Basophil detector, 23...Eosinophil detector, 24...
- Neutrophil detector, 26... comparator (nuclear corresponding signal extraction device). 31-52...Polar moment of inertia around the center of gravity) J calculation device, 53...Nuclear area A counter, 54 and 55
. . . R value calculation device, 56 to 60 . . . Discrimination means.

Claims (1)

[Claims] 1. Each part of the white blood cell sample divided into tiny quanta is two-dimensionally scanned in sequence by high-speed scanning in the Y direction and low-speed scanning in the X direction that intersects with this. a sample scanning device that detects the sample scanning signal; a device that detects that the white blood cells under test are neither basophils, eosinophils, nor neutrophils based on the sample scanning signal; A device that extracts a signal corresponding to the white plum bulb nucleus by wave height discrimination, a counter that obtains the nucleus area A by counting the nucleus corresponding signal, and a counter that obtains the nucleus area A by counting the nucleus corresponding signal, A device that calculates the moment of inertia J, a calculation device that calculates the above nuclear area A and polar moment of inertia) J, and when the white blood cell is not a basophil, an eosinophil, or a neutrophil. a discriminating means for discriminating the leukocytes as monocytes when the R value exceeds a value between 0.204 and 0.207 and discriminating the leukocytes as lymphocytes when the R value does not exceed the above value; Ball discriminator.