KR102122020B1 - Apparatus for analyzing blood cell, method for analyzing blood cell using the apparatus - Google Patents

Abstract

The blood cell analysis apparatus according to the present invention includes a first optical system composed of a first color sorting mirror reflecting a first laser incident from the first laser generating device in a first path and a first beam shaper arranged along the first path ; And a second color sorting mirror that transmits the second laser incident from the second laser generating device through the second path and reflects the side scattered light traveling in the second path among the side scattered light for the first laser, and the second path. It may include a second optical system composed of a second beam shaper arranged accordingly.

Description

Blood cell analysis device, blood cell analysis method using the same {APPARATUS FOR ANALYZING BLOOD CELL, METHOD FOR ANALYZING BLOOD CELL USING THE APPARATUS}

The present invention relates to a blood cell analysis device, a blood cell analysis method using the same, and more particularly, to a blood cell analysis device that analyzes a type of blood cell by irradiating a laser to a blood sample, and a blood cell analysis method using the same.

In flow cytometry or leukocyte differential test (CBC), various blood cells such as white blood cells are classified and used to classify normal cells and abnormal cells. These tests are used to test for various cell diseases, and it is very important to accurately classify multiple cells.

Among the methods of analyzing blood cells, there is a technique of classifying blood cells by measuring lasers scattered by blood cells by irradiating a laser to the blood cells. In this regard, it will be described with reference to FIG. 1.

1 is a conceptual diagram for explaining a blood cell analysis technique using a laser.

As shown in the figure, when a blood sample is flowed into the microtubule to analyze blood cells, and when the laser is irradiated to the microtubule in this process, the laser is irradiated to specific blood cells, such as forward scattered light and lateral scattered light ( side scattered light).

Among these, it is known that the forward scattered light is affected by the size of the cells, and the lateral scattered light is affected by the structure of the cells. Therefore, the type of blood cells can be distinguished by measuring the scattering angle and intensity of the laser, which varies depending on the type, size, and the like of blood cells.

Among the prior art, a blood cell analysis apparatus using such a laser irradiation method is disclosed, but since most of the cells are irradiated with a laser in a single direction, the density of light measured according to the laser irradiation angle varies, so that the cells cannot be accurately classified. There is this.

In addition, for accurate analysis, the laser and the scattered light must be transformed into a predetermined predetermined shape, and the conventional blood cell analysis apparatus changes each optical system to modify the shape of the laser light incident on the blood cells and the shape of the laser light scattered from the blood cells. Since it is necessary, there is a problem that the complexity of the device increases and the production cost increases.

Korean Registered Patent No. 10-1681422 Korean Patent Publication No. 10-2018-0051844

One aspect of the present invention is to irradiate a plurality of lasers at different angles to the blood sample, but it is possible to simplify the structure of the optical system by designing the same as the scattering path of one laser and the incident path of the other laser to the blood sample. A blood cell analysis device and a blood cell analysis method using the same are provided.

The technical problems of the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned will be clearly understood by those skilled in the art from the following description.

In the blood cell analysis apparatus for measuring light scattered by blood cells by irradiating a laser to a microtubule through which blood cells move according to an embodiment of the present invention, the first laser generated by the first laser generator The first color sorting mirror reflecting the first laser incident from the first laser generating device to the first path, and the reflected first laser in a predetermined cross-sectional shape so as to be irradiated along the first path to the customs. The second laser, which is generated by the first optical system and the second laser generating device, which is composed of the first beam shaper arranged along the first path, to be irradiated to the customs, and has a wavelength higher than the wavelength of the first laser, is The second path of the lateral scattered light to the first laser irradiated to the blood cells while transmitting the second laser incident from the second laser generator to the second path to be irradiated along the second path to the customs A second color sorting mirror reflecting the lateral scattered light traveling to and the second laser and the scattered light of the first laser traveling through the second path have a predetermined cross-sectional shape, along the second path And a second optical system composed of second beam shapers arranged.

The first beam shaper and the second beam shaper include a beam expander, a cylinder lens, and a condenser lens, wherein the first path or the second path from the microtubule Accordingly, the beam expander, the cylinder lens, and the focusing lens may be optical systems sequentially arranged.

The first color sorting mirror is disposed at the rear end on the first path from the focusing lens of the first beam shaper, and the second color sorting mirror is disposed at the rear end on the second path from the focusing lens of the second beam shaper. Can be.

The predetermined cross-sectional shape may be an ellipse whose major axis is larger than the diameter of the microtube, and its minor axis is smaller than the size of the blood cells.

The first color-coded mirror, the first laser incident from the first laser generating device to separate the side scattered light for the first laser and the second laser superimposed on the first path, the first laser While reflecting in the path, lateral scattered light to the second laser incident along the first path can be transmitted.

The second color-coded mirror, the second laser is incident on the second path from the second laser generating device, so as to separate the scattered light for the second laser and the first laser superimposed on the second path 2 While transmitting the laser, lateral scattered light to the first laser incident along the second path may be reflected.

The blood cell analysis device is disposed on a straight line extending from the first path, and includes a first front light detector for measuring front scattered light for the first laser irradiated to the blood cells; A second forward light detector arranged on a straight line extending from the second path to measure forward scattered light for the second laser irradiated to the blood cells, and lateral scattered light for the first laser irradiated to the blood cells The first lateral light detector for measuring lateral scattered light traveling in the second path and a first lateral scattered light disposed on a straight line extending from the first path and irradiating the blood cells to the second laser A second lateral light detector for measuring lateral scattered light traveling through the path may be further included.

The first side light detector and the second side light detector are photo-multiplier tubes (PMT), and the first front light detector and the second front light detector are photo-diodes (PD). Can be

A third color selection mirror disposed between the first color selection mirror and the second lateral light detector, and reflecting fluorescence traveling in the first path among the fluorescence for the second laser irradiated to the blood cells and the A fluorescence detector measuring fluorescence reflected by the third color selection mirror may be further included.

The third color sorting mirror may transmit lateral scattered light to the second laser incident along the first path and reflect fluorescence from the second laser incident along the first path.

The fluorescence detector may be a photo-multiplier tube (PMT).

The first path and the second path may be orthogonal paths on the same two-dimensional plane.

At least one first disposed between the first laser generating device and the first color sorting mirror to reflect the first laser generated by the first laser generating device to be incident on the first color sorting mirror. At least one disposed between the reflection mirror and the second laser generating device and the second color sorting mirror to reflect the second laser generated by the second laser generating device to be incident on the second color selecting mirror. The second reflection mirror of the may further include.

In addition, in the blood cell analysis method using the blood cell analysis apparatus according to an embodiment of the present invention, the first laser and the second laser having different angles of incidence are irradiated to the microtubule through which blood cells move, and the first path is followed. The second laser irradiated to the microtube along the same path as the side scattered light traveling along the second path among the side scattered light for the first laser, and measuring the front scattered light and the side scattered light for the irradiated first laser Measures the forward scattered light and the lateral scattered light for, the forward scattered light for the first laser running in the direction extending from the measured first path, the lateral scattered light for the first laser traveling along the second path, The type of the blood cells is analyzed based on the forward scattered light for the second laser traveling in the direction extending from the second path and the lateral scattered light for the second laser traveling along the first path.

Further comprising measuring the fluorescence proceeding along the first path among the fluorescence for the second laser irradiated with the blood cells, and analyzing the type of the blood cells, the front for the measured first laser Analyze the type of the blood cells based on the voltage magnitude of scattered light, lateral scattered light for the first laser, forward scattered light for the second laser, lateral scattered light for the second laser, and fluorescence for the second laser It may be.

According to one aspect of the present invention described above, one laser incident on the blood sample and the other lasers scattered by the blood sample are designed to have the same travel path, and the optical beams of different lasers are the same. It can be formed, and by analyzing the blood cells based on a plurality of scattered light for the laser irradiated at different angles, it is possible to ensure the reliability of the blood cell analysis results.

1 is a conceptual diagram for explaining a blood cell analysis technique using laser irradiation.
2 is a view showing the configuration of the blood cell analysis apparatus according to an embodiment of the present invention.
FIG. 3 is a view showing a movement path of scattered light for the first laser and the second laser irradiated by the blood cell analyzer of FIG. 2.
4 is a view showing an example in which the cross-sectional shape of the laser is changed by the beam shaper.
5 is a diagram illustrating an example in which a detection result is changed according to a shape of a laser cross-section irradiated with a micro tube.
6 is a view showing an example of a cross-sectional shape of the side scattered light incident on the side light detector.
7 is a view showing the configuration of the blood cell analysis apparatus according to another embodiment of the present invention.
8 is a flowchart illustrating a schematic flow of a blood cell analysis method according to an embodiment of the present invention.
9 and 10 are graphs showing signal measurement results for blood cell analysis using the blood cell analysis apparatus according to the present invention according to the present invention.

For a detailed description of the present invention, which will be described later, reference is made to the accompanying drawings that illustrate, by way of example, specific embodiments in which the present invention may be practiced. These examples are described in detail enough to enable those skilled in the art to practice the present invention. It should be understood that the various embodiments of the present invention are different, but need not be mutually exclusive. For example, the specific shapes, structures, and properties described herein can be implemented in other embodiments without departing from the spirit and scope of the invention in connection with one embodiment. In addition, it should be understood that the location or placement of individual components within each disclosed embodiment can be changed without departing from the spirit and scope of the invention. Therefore, the following detailed description is not intended to be taken in a limiting sense, and the scope of the present invention, if appropriately described, is limited only by the appended claims, along with all ranges equivalent to those claimed. In the drawings, similar reference numerals refer to the same or similar functions throughout several aspects.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to the drawings.

2 is a view showing a blood cell analysis apparatus according to an embodiment of the present invention.

The blood cell analysis apparatus 1 according to an embodiment of the present invention applies pressure to the microtube T to allow the blood sample to move at a high speed inside the microtube T, and at different angles to the moving blood sample. It is a device that classifies the type of blood cells included in a blood sample by irradiating a plurality of lasers, measuring a plurality of scattered light for the irradiated laser, and analyzing the measured scattered light.

Specifically, the blood cell analysis apparatus 1 according to an embodiment of the present invention includes a plurality of laser generators 110 and 120, beam shapers 510 and 520, and color selection mirrors 210 and 220. Optical system (710, 720), a plurality of front light detectors (310, 320) and a plurality of side light detectors (410, 420).

The plurality of laser generating devices 110 and 120 are installed around the microtube T through which blood cells move, and are light source light emitting devices that irradiate the blood sample flowing inside the microtube T with a laser.

The blood cell analyzing apparatus 1 according to the present invention is provided with at least two laser generating devices 110 and 120, and the first laser generating device 110 and the second laser generating device 120 are different incidences into blood samples. It can be arranged to irradiate the laser at an angle. In the illustrated embodiment, two laser generating devices 110 and 120 are represented, but three or more laser generating devices may irradiate the laser at different angles depending on a measurement environment and precision of a required analysis result. have.

In addition, the first laser irradiated from the first laser generator 110 and the second laser irradiated from the second laser generator 120 may have different center wavelengths. For example, the first laser generator 110 may irradiate a first laser having a central wavelength of 488 nm, and the second laser generator 120 may irradiate a second laser having a central wavelength of 635 nm. However, the center wavelengths of the first laser and the second laser are not limited to the above-described embodiments, and the numerical values are not limited as long as the center wavelengths of the first laser and the second laser are different.

The first optical system 710 and the second optical system 720 are installed in the first path and the second path, while transmitting or reflecting the laser traveling in the installed path, while the laser traveling in the path has a predetermined cross-section. I can do it.

The first optical system 710 includes a first beam shaper 510 and a first color sorting mirror 210, and the second optical system 720 includes a second beam shaper 510 and a second color sorting mirror 220. It is composed of an optical system.

A plurality of color sorting mirrors (dichroic mirrors 210, 220) are planar mirrors coated with a transparent multi-layer thin film, reflecting light in a specific wavelength range due to the interference effect of light in the thin film, and light of other wavelengths Is an optical member that transmits.

In this case, the color selection mirrors 210 and 220 according to an embodiment of the present invention are characterized in that the first laser having the first center wavelength reflects and the second laser having the second center wavelength transmits. For example, when the first laser generator 110 irradiates a first laser having a central wavelength of 488 nm, and the second laser generator 120 irradiates a second laser having a central wavelength of 635 nm, color The screening mirror may be designed to have a characteristic of passing only wavelengths in a band of 600 nm or more. Accordingly, while the first laser is reflected by the color selection mirror, the second laser can pass through the color selection mirror.

Among the plurality of color sorting mirrors 210 and 220, the first color sorting mirror 210 is disposed around a position where the first laser generating device 110 is disposed, and the second color sorting mirror 220 generates a second laser The device 120 may be disposed around the position in which it is disposed.

The first color sorting mirror 210 reflects the first laser irradiated from the first laser generator 110 so that the first laser passes the first path (for example, 12 o'clock based on the microtube T). Accordingly, blood samples flowing inside the microtube T may be irradiated. In other words, the first color sorting mirror 210 may reflect the first laser incident in any direction in the first path.

The second color sorting mirror 220 reflects the second laser incident from the second laser generator 120 so that the second laser passes the second path (eg, at 3 o'clock based on the microtube T). Accordingly, blood samples flowing inside the microtube T may be irradiated.

On the other hand, at least one reflective mirror (R1, R2) is installed on the first laser generator 110 and the second laser generator 120, the first laser and the first irradiated from the first laser generator 110 2 The second laser irradiated from the laser generator 120 may be incident on the first color sorting mirror 210 and the second color sorting mirror 220 at a predetermined angle (for example, 45°).

As such, angles at which the first laser and the second laser are irradiated to the blood cells may be determined according to the placement position and the installation angle of the color selection mirrors 210 and 220. In this embodiment, the placement position and installation angle of the color selection mirrors 210 and 220 are such that the first path through which the first laser is incident and the second path through which the second laser is incident have an orthogonal path on a two-dimensional plane. Can be set. However, the illustrated first path and the second path do not necessarily have to be orthogonal, and the first path and the second path are designed to have an obtuse or acute angle according to the arrangement position and installation angle of the color sorting mirrors 210 and 220. It may be.

A plurality of beam shapers (beam shaper, 510, 520) is an optical system that transforms the incident laser into a predetermined shape. Specifically, the beam shapers 510 and 520 are optical systems composed of beam expanders 511 and 521, cylinder lenses 512 and 522, and condenser lenses 511 and 521. The beam shaper 510 may be disposed on the first path, and the second beam shaper 520 may be disposed on the second path.

Specifically, in the first beam shaper 510, the first focusing lens 511, the first cylinder lens 512, and the first beam expander 513 are sequentially arranged along the first path from the microtube T. Optical system, the second beam shaper 520 is a second focusing lens 521, a second cylinder lens 522 and a second beam expander 523 are sequentially arranged along the second path from the microtube T It may be an optical system.

In this case, the first color sorting mirror 210 is installed at the rear end of the first beam expander 513, and the first beam shaper 510 installed on the first path is the first laser incident from the first laser generator 110. ), the first laser may sequentially pass through the first beam expander 513, the first cylinder lens 512, and the first focusing lens 511. Similarly, the second color sorting mirror 220 may transmit the second laser to sequentially pass through the second beam expander 523, the second cylinder lens 522, and the second focusing lens 521. .

The front light detectors 310 and 320 and the side light detectors 410 and 420 are photoelectric conversion elements that convert the incident light into current. That is, the front light detectors 310 and 320 and the side light detectors 410 and 420 may represent the characteristics of the sensed light by converting the converted current into a voltage output proportional to the current intensity.

The plurality of forward light detectors 310 and 320 may measure forward scattered light for a laser irradiated to blood cells flowing through the microtube T. In addition, the plurality of side light detectors 410 and 420 may measure side scattered light for a laser irradiated to blood cells. In this regard, it will be described with reference to FIG. 2 together.

FIG. 3 is a view showing a movement path of scattered light for the first laser and the second laser.

Among the plurality of front light detectors 310 and 320, the first front light detector 310 measures the forward scattered light for the first laser irradiated to the blood cells, and the second front light detector 320 is irradiated to the blood cells The forward scattered light for the second laser can be measured.

To this end, the first front light detector 310 may be disposed at a position spaced a predetermined distance from the opposite side of the incident surface of the microtube T into which the first laser is incident. Specifically, the first front light detector 310 is a straight line extending from the first path, which is the path where the first laser is irradiated with blood cells, so that the front scattered light scattered in the same or similar direction as the first path can be incident. Can be placed on. For example, when the first laser irradiates blood cells in the 12 o'clock direction of the microtubule, the first anterior photodetector 310 may be disposed at 6 o'clock.

Similarly, the second front light detector 320 may be disposed at a position spaced a predetermined distance from the opposite side of the incident surface of the microtube T in which the second laser is incident, and specifically, the second laser may be used as blood cells. It may be disposed on a straight line extending from the irradiated second path.

On the other hand, the first lateral light detector 410 of the plurality of lateral light detectors 410 and 420 measures lateral scattered light for the first laser irradiated to the blood cells, and the second lateral light detector 420 is attached to the blood cells. Lateral scattered light for the irradiated second laser can be measured.

In this case, the first side light detector 410 may measure the side scattered light traveling in the second path, which is the path through which the second laser is incident, among the side scattered lights in all directions to the first laser.

When the first laser is irradiated with blood cells, scattered light is generated in a plurality of directions depending on the type of blood cells. As some of them progress along the second path, lateral scattered light traveling along the second path may be incident on the second color selection mirror 220 disposed on the second path. As described above, the second color sorting mirror 220 may reflect the side scattered light for the first laser having the first center wavelength incident along the second path, and the first side light detector 410 may One of the side scattered light for the laser may be disposed on the reflection path of the side scattered light of the first laser reflected by the second color sorting mirror 220 so as to measure the side scattered light traveling in the second path.

Similarly, the second lateral light detector 420 may measure lateral scattered light traveling in a first path, which is a path in which the first laser is incident, among lateral scattered lights in all directions to the second laser. To this end, the second side light detector 420 may be disposed on the reflection path of the side scattered light of the second laser reflected by the first color sorting mirror 210.

Here, the front photo detectors 310 and 320 are photo-diodes (PDs), and the side photo detectors 410 and 420 may be provided as photo-multiplier tubes (PMTs).

In general, since the laser has a strong linearity, forward scattered light scattered in a straight line extending from the incident path is easy to measure. However, lateral scattered light, which is laser light scattered side by side by blood cells, is relatively weak compared to forward scattered light. Accordingly, the side light detectors 410 and 420 may be provided as a photoelectric amplification tube equipped with transimpedence to amplify the sensed signal to effectively detect weak light. However, the present invention is not limited thereto, and both the front light detectors 310 and 320 and the side light detectors 410 and 420 may be provided as photoelectric diodes or photoelectric amplification tubes.

As described above, the blood cell analysis apparatus 1 according to an embodiment of the present invention is different from the prior art in that the light path through which one laser is irradiated to the blood cells and the different wavelengths generated in the blood cells by the other lasers By allowing the scattered light paths to coincide, different lasers can be transformed into the same beam shape with one optical system (first and second beam shapers 510 and 520). In this regard, it will be described with reference to FIGS. 4 to 6 together.

FIG. 4 is a diagram illustrating an example in which the cross-sectional shape of a laser is changed by a beam shaper, and FIG. 5 is a diagram showing an example in which a detection result is changed according to a laser cross-sectional shape irradiated to a microtube, and FIG. 6 is a laser An example of a cross-sectional shape is shown.

As described above, a part of the side scattered light for the first laser irradiated from the first laser generator 110 and the second laser irradiated by the second laser generator 120 proceeds along the first path. , A portion of the second laser irradiated from the second laser generator 120 and the side scattered light with respect to the first laser may proceed along the second path.

Accordingly, the first beam shaper 510 disposed on the first path may allow the side scattered light for the first laser and the second laser traveling along the first path, which is the same path, to have the same beam shape. In addition, the second beam shaper 520 may allow the lateral scattered light for the second laser and the first laser traveling along the second path, which is the same path, to have the same beam shape.

As illustrated in FIG. 4, the first and second lasers generated by the laser generators 110 and 120 are reflected or transmitted through the color sorting mirrors 210 and 220 and beam expanders of the beam shapers 510 and 520 ( 513, 523). The laser that has passed through the beam expanders 513 and 523 may be incident on the cylinder lenses 512 and 522 in a state in which the cross section is expanded, and the vertical axis (short axis) diameter is reduced while passing through the cylinder lenses 512 and 522. It can be transformed into a shape.

Thereafter, when a laser having an elliptical cross-section is incident on the focusing lenses 511 and 521, the cross-sectional area may be reduced as a whole. Specifically, the laser passing through the beam shapers 510 and 520 may be deformed such that the long axis has an elliptical cross section larger than the diameter of the microtube T and the short axis smaller than the size of the blood cells.

That is, the first laser passing through the first optical system 710 and the second laser passing through the second optical system 720 may have a cross-sectional shape as shown in (a) of FIG. 5.

At this time, when the laser cross-sectional shape irradiated to the microtube T is as shown in FIG. Because it is not scattered, blood cells may not be detected. In addition, when the laser cross-sectional shape irradiated to the microtubule T is shorter than the size of the blood cells, as shown in FIG. 5(c), two blood cells that move continuously are linked to one blood cell. As the result can be measured, the accuracy of the analysis may be deteriorated. On the other hand, when the laser cross-sectional shape has a shape as shown in Figure 5 (a), it is possible to not only detect blood cells flowing into the entire region of the microtubule (T), but also continuously flow blood cells into individual blood cells. It has an effect that can be detected.

As described above, the laser generated by the laser generating devices 110 and 120 includes beam expanders 513 and 523 and cylinder lenses 512 and 522 of the beam shapers 510 and 520 disposed in the first path or the second path. And while passing through the focusing lenses (511, 521) sequentially, the long axis is larger than the diameter of the microtube (T), the minor axis is to be irradiated to the microtube (T) in a deformed state to have an elliptical cross-section smaller than the size of the blood cells Can be.

In addition, in the blood cell analysis apparatus 1 according to an embodiment of the present invention, a distance between components constituting the optical system may be designed such that the laser has a predetermined cross-sectional shape. For example, the distance between the microtubule T and the focusing lenses 511 and 521 is 7 mm, the distance between the focusing lenses 511 and 521 and the cylinder lenses 512 and 522 is 23 mm, the cylinder lenses 512 and 522 and the beam expander. When the optical system is designed such that the distance between 513 and 523 is 25 mm, and the distance between the beam expanders 513 and 523 and the color sorting mirrors 210 and 220 is 40 mm, the laser may have the above-described cross-sectional shape. However, the distance between the components constituting the optical system is not limited to the above-described examples, and the distance between the components may be changed according to the diameter of the microtube T and the type of blood cells to be analyzed.

On the other hand, the scattered light of the first laser and the second laser irradiated to the blood cells may proceed in a direction opposite to the laser irradiated to the microtube (T). That is, the lateral scattered light of the first laser traveling along the second path passes through the second beam shaper 520 in the order of the second focusing lens 521, the second cylinder lens 522, and the second beam expander 523. The lateral scattered light of the first laser that has passed through the second beam expander 523 may be reflected on the second color sorting mirror 220 in a state in which the cross-section is expanded and enter the first lateral light detector 410. have. At this time, a focusing lens (not shown) for reducing the cross-sectional area of the side scattered light of the first laser may be additionally disposed between the second color selection mirror 220 and the first side light detector 410, and accordingly, the first color The side light detector 410 may detect side scattered light having a cross-sectional shape as shown in FIG. 6. That is, as the side scattered light passes through the beam shapers 510 and 520 in the reverse order, the final beam shape may have a shape in which the horizontal axis is short and the vertical axis is long as compared to FIG. 5.

Accordingly, the laser shape required by the incidence unit (micro tube (T)) and the laser shape required by the measurement unit (front light detectors 310 and 320 and side light detectors 410 and 420) are satisfied with one optical system. I can do it.

In this process, the first color sorting mirror 210 and the second color sorting mirror 220 may separate two lasers having different wavelengths traveling in the same path.

That is, the first color sorting mirror 210 firstly applies the first laser incident from the first laser generator 110 to separate the side scattered light for the first laser and the second laser superimposed on the first path. While reflecting in the path, lateral scattered light to the second laser incident along the first path can be transmitted. In addition, the second color sorting mirror 210 separates the second laser from the second laser generating device 120 so as to separate the scattered light for the second laser and the first laser superimposed on the second path. While transmitting through the path, lateral scattered light to the first laser incident along the second path can be reflected.

Accordingly, the second side light detector 420 disposed on a straight line extending from the first path can detect only the side scattered light of the second laser selectively transmitted by the first color sorting mirror 210, and The one side light detector 410 can detect only the side scattered light of the first laser that is selectively reflected by the second color sorting mirror 220 to extract the exact characteristics of each side scattered light.

In some other embodiments, the blood cell analysis apparatus according to the present invention may be further provided with a means for measuring fluorescence. In this regard, it will be described with reference to FIG. 4.

7 is a view showing a blood cell analysis apparatus according to another embodiment of the present invention.

Specifically, the blood cell analysis device 2 according to another embodiment of the present invention includes a plurality of laser generating devices 110 and 120, a plurality of color sorting mirrors 210, 220 and 230, and a plurality of front light detectors 310, 320), a plurality of side light detectors (410, 420), a plurality of beam shapers (510, 520) and a fluorescence detector (600).

At this time, the first laser generating device 110, the second laser generating device 120, the first color sorting mirror 210, the second color sorting mirror 220, the first front light detector 310, the second front Since the photo detector 320, the first side photo detector 410, the second side photo detector 420, the first beam shaper 510, and the second beam shaper 520 are the same as those shown in FIG. , Repeated description will be omitted.

That is, the blood cell analysis apparatus 2 according to another embodiment of the present invention includes a third color selection mirror 230 and a fluorescence detector 600 on the blood cell analysis apparatus 20 according to an embodiment of the present invention shown in FIG. 1. ) May be further provided.

The third color sorting mirror 230 may be disposed between the first color sorting mirror 210 and the second side light detector 420. The third color sorting mirror 230 may reflect fluorescence among lateral scattered light of the second laser transmitted from the first color sorting mirror 210.

The side scattered light for the second laser transmitted by the first color sorting mirror 210 may include a wavelength (the phenomenon in which the frequency is modulated by cells). In this case, the blood cell analysis apparatus 1 shown in FIG. 1 uses a band pass filter on the side of the second side light detector 420 to prevent the fluorescent laser light from being detected by the second side light detector 420. It can be equipped to block the fluorescent laser.

On the other hand, the blood cell analysis apparatus 2 according to another embodiment of the present invention is further provided with a third color selection mirror 230, and transmitted through the first color selection mirror 210 by the third color selection mirror 230. While transmitting lateral scattered light to the general second laser, fluorescence from the second laser incident along the first path may be reflected.

The fluorescence detector 600 may measure fluorescence reflected by the third color sorting mirror 230. To this end, the fluorescence detector 600 may be disposed on a reflection path of fluorescence reflected by the third color selection mirror 230, but may be in the form of an optoelectronic amplifying tube, but is not limited thereto.

As such, the blood cell analysis apparatus 2 according to another embodiment of the present invention can additionally collect measurement information for a fluorescence channel, thereby improving accuracy of blood cell analysis.

8 is a flowchart illustrating a schematic flow of a blood cell analysis method according to an embodiment of the present invention.

The blood cell analysis method according to the present embodiment may be performed by the blood cell analysis devices 1 and 2 according to the embodiments of the present invention. Hereinafter, for convenience of description, the blood cell analysis method according to an embodiment of the present invention will be described on the assumption that it is performed by the blood cell analysis apparatus 1 shown in FIG. 1.

The blood cell analysis apparatus 1 may drive the first laser generator 110 and the second laser generator 120 to irradiate the first laser and the second laser to the microtube T (51).

At this time, the first laser and the second laser may have different incidence angles, the first laser is reflected by the first color sorting mirror 210 and is incident along the first path, and the second laser is the second color Transmitted by the selection mirror 220 may be incident along the second path orthogonal to the first path.

The blood cell analyzer 1 may measure front scattered light and lateral scattered light for the first laser, and front scattered light and lateral scattered light for the second laser (53).

Specifically, the first front light detector 310 provided in the blood cell analyzer 1 measures the front scattered light for the first laser, and the second front light detector 320 measures the front scattered light for the second laser. The first side light detector 410 measures side scattered light for the first laser, and the second side light detector 420 measures side scattered light for the second laser.

At this time, the blood cell analysis apparatus 1 according to an embodiment of the present invention measures the lateral scattered light of the second laser traveling along the first path, which is the incident path of the first laser, and the second, which is the incident path of the second laser The positions of the first side light detector 410 and the second side light detector 420 may be designed to measure the side scattered light of the first laser traveling along the path.

In addition, beam shapers 510 and 520 are provided on the first path and the second path so that beam shapes of different lasers overlapping on the first path and the second path can be transformed using one optical system. Can be. At the same time, in order to separate different lasers overlapping on the first path and the second path, lasers having a first center wavelength on each path are reflected, while lasers having a second center wavelength greater than the first center wavelength The color selection mirrors 210 and 220 to pass may be disposed.

Thereafter, the blood cell analyzer 1 may analyze the type of blood cell based on the measured plurality of scattered lights (55). In this regard, it will be described with reference to FIGS. 9 and 10 together.

9 and 10 are graphs showing signal measurement results for blood cell analysis using the blood cell analysis device 1 according to the present invention.

9 is a graph showing a signal measurement result for a first white blood cell sample, and FIG. 10 is a graph showing a signal measurement result for a second white blood cell sample.

According to a component such as a nucleus constituting a cell, when the laser light is irradiated to the cell, the light is transmitted, refracted, reflected or fluorescent based on the cell to be classified. Using this effect, the distribution of laser light, that is, the scattering angle and the amount of light of the first laser and the second laser are measured and compared with pre-stored reference information to classify blood cells.

The illustrated figure is a result of measuring a signal for a white blood cell sample through erythrocyte lysis in whole blood. Each scattered light is measured by a voltage signal having a different size, but it can be seen that the measured time points are almost similar. That is, as the laser is irradiated to the blood cells, the laser transformed from the original laser is scattered, and the blood cell analyzer 1 analyzes the type of blood cells detected based on the variation and characteristics of the voltage measured through the photo detector. Can be.

Although described above with reference to embodiments, those skilled in the art understand that various modifications and changes can be made to the present invention without departing from the spirit and scope of the present invention as set forth in the claims below. Will be able to.

1, 2: blood cell analysis device
110: first laser generator
120: second laser generating device
210: first color sorting mirror
220: second color sorting mirror
230: third color sorting mirror
310: first front light detector
320: second front light detector
410: first side light detector
420: second side light detector
510: first beam shaper
520: second beam shaper
600: fluorescence detector

Claims (15)

In the blood cell analysis apparatus for measuring the light scattered by blood cells by irradiating a laser to the microtubule through which blood cells move,
A first color sorting mirror reflecting the first laser incident from the first laser generator to the first path so that the first laser generated by the first laser generator is irradiated along the first path to the microtube. , A first optical system composed of a first beam shaper arranged along the first path so that the reflected first laser is irradiated to the microtube in a predetermined cross-sectional shape; And
The second laser incident from the second laser generator, such that a second laser having a wavelength higher than that of the first laser is irradiated along the second path to the microtube, generated by the second laser generator. A second color selection mirror reflecting lateral scattered light traveling through the second path of the lateral scattered light to the first laser irradiated to the blood cells while transmitting the second path, and the transmitted second laser and the And a second optical system composed of a second beam shaper arranged along the second path such that the scattered light of the first laser traveling in the second path has a predetermined cross-sectional shape,
The first color selection mirror,
The first laser incident from the first laser generator is reflected to the first path while separating lateral scattered light for the first laser and the second laser superimposed on the first path, while the first laser is reflected. A blood cell analysis device that transmits lateral scattered light to the second laser incident along a path.
According to claim 1,
The first beam shaper and the second beam shaper,
Includes beam expanders, cylinder lenses, and condenser lenses,
A blood cell analysis device, wherein the beam expander, the cylinder lens, and the focusing lens are sequentially arranged along the first path or the second path from the microtube.
According to claim 2,
The first color sorting mirror is disposed at a rear end on the first path from the focusing lens of the first beam shaper,
The second color sorting mirror is disposed on the rear end on the second path from the focusing lens of the second beam shaper, a blood cell analysis device.
According to claim 2,
The predetermined cross-sectional shape is a blood cell analysis device, the major axis of which is larger than the diameter of the microtube, and the minor axis is smaller than the size of the blood cells.
delete According to claim 1,
The second color sorting mirror,
The second laser incident on the second path is transmitted from the second laser generator to transmit the second laser and the second laser superimposed on the second path. 2 A blood cell analysis device that reflects side scattered light to the first laser incident along the path.
According to claim 1,
A first forward photodetector disposed on a straight line extending from the first path to measure forward scattered light for the first laser irradiated to the blood cells; A second front light detector arranged on a straight line extending from the second path to measure front scattered light for the second laser irradiated to the blood cells;
A first lateral light detector measuring lateral scattered light traveling in the second path among lateral scattered light to the first laser irradiated to the blood cells; And
It is disposed on a straight line extending from the first path, further comprising a second side light detector for measuring the side scattered light traveling to the first path of the side scattered light for the second laser irradiated to the blood cells, Blood cell analysis device.
The method of claim 7,
The first side light detector and the second side light detector are photo-multiplier tubes (PMT),
The first anterior photodetector and the second anterior photodetector are photo-diode (PD) devices.
The method of claim 7,
A third color sorting mirror disposed between the first color sorting mirror and the second lateral light detector to reflect fluorescence traveling in the first path among fluorescence on the second laser irradiated to the blood cells; And
And a fluorescence detector for measuring fluorescence reflected by the third color selection mirror.
The method of claim 9,
The third color selection mirror,
A blood cell analysis device that transmits lateral scattered light to the second laser incident along the first path and reflects fluorescence from the second laser incident along the first path.
The method of claim 9,
The fluorescence detector is a photoelectron amplification tube (Photo-Multiplier Tube, PMT), blood cell analysis device.
According to claim 1,
The first path and the second path are orthogonal paths on the same two-dimensional plane, a blood cell analysis apparatus.
According to claim 1,
At least one first disposed between the first laser generating device and the first color selection mirror to reflect the first laser generated by the first laser generation device to be incident on the first color selection mirror. Reflective mirrors; And
At least one second disposed between the second laser generating device and the second color selection mirror to reflect the second laser generated by the second laser generation device to be incident on the second color selection mirror. A blood cell analysis device further comprising a reflection mirror.
In the blood cell analysis method using the blood cell analysis device according to claim 1,
The first laser and the second laser having different angles of incidence are irradiated to the microtubule through which blood cells move,
The forward scattered light and the lateral scattered light for the first laser irradiated along the first path are measured, and the microtube is irradiated along the same path as the lateral scattered light traveling along the second path among the lateral scattered light for the first laser. Measuring the front scattered light and the side scattered light for the second laser,
Measured forward scattering light for the first laser running in a direction extending from the first path, lateral scattering light for the first laser running along the second path, and running in a direction extending from the second path A blood cell analysis method for analyzing the type of the blood cells based on the forward scattered light for the second laser and the lateral scattered light for the second laser traveling along the first path.
The method of claim 14,
Further comprising measuring the fluorescence proceeding along the first path among the fluorescence for the second laser irradiated with the blood cells,
Analyzing the type of blood cells,
Measure the voltage magnitude of each of the measured forward scattered light for the first laser, lateral scattered light for the first laser, forward scattered light for the second laser, lateral scattered light for the second laser, and fluorescence for the second laser. A blood cell analysis method for analyzing the types of the blood cells based on the basis.

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