KR20030032809A - Apparatus for diagnosing disease by detecting properties of red blood cells and diagnosing method thereof - Google Patents

Abstract

PURPOSE: An apparatus and a method for diagnosing disease using physical characteristics of erythrocyte are provided to detect easily the physical characteristics of erythrocyte by improving a structure of a micro chip and a structure of an optical detection member. CONSTITUTION: A micro chip(210) is fixed to a fixing plate(230). An optical detection member(220) is arranged on an upper portion of the micro chip(210). The optical detection member(220) is formed with a microscope or a lens array. A charge coupled device(255) is connected to the optical detection member(220). A frame grabber(250) is connected to the charge coupled device(255). An analysis device(260) is used for analyzing an obtained image and outputting an analyzed result. An optical generation member(245) is arranged at a lower portion of the fixing plate(230) in order to irradiate the light to the micro chip(210). A fixing member(240) is formed at one side of the fixing plate(230).

Description

Apparatus for diagnosing disease by detecting properties of red blood cells and diagnosing method according to physical properties of red blood cells

The present invention relates to a device for diagnosing a disease using a characteristic of a red blood cell and a diagnostic method thereof, and more particularly, by easily detecting a physical characteristic of a red blood cell included in blood by including an improved microchip and optical detection means. The present invention relates to a disease diagnosis apparatus using the physical characteristics of red blood cells capable of diagnosing various diseases related to red blood cells, and a diagnostic method thereof.

In general, red blood cells, which are one of the constituents of human blood together with white blood cells and platelets, have a donut shape with a clogged center, and have a diameter of about 8 to 12 μm and a thickness of about 1 to 2 μm, but are very flexible. This allows them to pass through narrow filters such as capillaries at high speed. As such, information about the erythrocytes, such as the degree, shape, and rate of passage of the erythrocytes as they deform during the passage of the filter, is used to diagnose hematological diseases. For example, information about erythrocytes generally relates to the number of red blood cells per unit volume (RBC) in the blood, the amount of hemoglobin per unit volume, the volume of erythrocytes, and the concentration of hemoglobin per erythrocyte. .

Currently, many studies have demonstrated that the RBC deformability of red blood cells, which is a characteristic of changing the shape of red blood cells under stress, is associated with necrosis of cells that develop diseases such as cancer. Cohen, for example, transplanted L1210 leukemia cells and Lewis lung carcinoma cells into experimental mice in the "Influence of tumor burden on red cell deformability by tumor growth", resulting in cancer cell death and reduced red blood cell deformability. And Dintefass reported that "Dome aspects of hemorheology of metastasis in malignant melanoma" increased blood viscosity and decreased red blood cell deformability with respect to melanoma, and Sevick and Jain reported that "Effect Of red blood cell rigidity on tumor blood flow: increase in viscous rigidity during hyperglycemia.

In addition, in Korea, Park Seok-won's "Study on Blood Viscosity and Erythrocyte Deformation in Patients with Malignant Tumors" measured the filtration time of red blood cells and compared the blood of control blood, urethral cancer patients, and gastric cancer patients. The difference was statistically confirmed. Also, the result of using the filtering method to measure the erythrocyte deformability through Oh Do Hoon's "Influence of Ginkgo biloba extract on the deformability of mouse erythrocytes." The average filtering time is about 11.8 seconds, compared to 33.1 seconds in the case of cancer mice.

As described above, the deformability of red blood cells is deeply associated with diseases such as cancer, and the study of the deformability of these red blood cells allows for early diagnosis of cancer as well as blood-related diseases such as diabetes and malaria. Is being studied.

On the other hand, International Patent Application Publication No. 93-701749 (name of the blood test apparatus of the examinee) proposes a blood test apparatus that can measure the number of red blood cells, white blood cells or platelets in the blood and diagnose the disease based thereon. It is. However, since the blood test apparatus disclosed in the international patent determines the presence or absence of a disease by measuring only the number of red blood cells and the like, it is difficult to determine a disease related to deformability of red blood cells.

In addition, US Pat. No. 5,798,827 to Robert S. Frank et al. Discloses a device capable of determining the shape of red blood cells.

1 shows a schematic diagram of a device capable of determining the shape of red blood cells in a fluid disclosed in U.S. Patent No. 5,798,827.

Referring to FIG. 1, the device for determining the shape of red blood cells includes a fluid cell 15 attached to the fluid system 10 and through which a diluted red blood cell sample passes. In the fluid cell 15, a measurement opening 20 is formed to pass red blood cells one by one, and electrodes 25 for connecting an electric power source 30 to both sides of the measurement opening 20 to form an electric field in the measurement opening 20. ) Is provided. The measurement opening 20 is arranged to measure the light scattered by the red blood cells passing therein.

The light source 35 is arranged to irradiate the light to the measurement opening 20, and is disposed to face the light source 35, and to the detection means 50 including the electrical resistance detector 40 and the scattered light detector 45. By this, light emitted from the light source 35 and scattered from the red blood cells is detected. The electrical resistance detector 40 and the scattered light detector 45 are respectively connected to a computer 55 having a digitizing means 60, a recording means 65 and a calculation means 70. The computer 55 converts the electric and optical signals scattered by the red blood cells into digital signals, and determines the shape of each red blood cell based on this.

However, the above-described apparatus for determining the morphology of erythrocytes, although the modified erythrocytes can be identified by diagnosing the morphology of erythrocytes, and the related diseases can be diagnosed, the deformability of erythrocytes cannot be measured. There is a problem that is difficult to judge about the disease involved.

On the other hand, US Patent No. 6,187,592, issued to Paul L. Gourley, is provided with a device for measuring the characteristics of red blood cells.

FIG. 2 is a schematic diagram of a device for measuring the properties of red blood cells shown in US Pat. No. 6,187,592, and FIG. 3A is a cross-sectional view of a microcavity part of the device for measuring the properties of red blood cells, and FIG. It is a top view of the microcavity shown to 3a.

Referring to FIG. 2, the apparatus for measuring the characteristics of the red blood cells includes an upper mirror 80, a lower mirror 85, an acquisition medium 90, and an analysis region 95, in which the red blood cells 105 are located. Has a resonant optical cavity 100 to be analyzed.

A laser pump 115 is arranged to activate the acquisition medium 90, and the light beam 110 generated from the laser pump 115 passes through the beam splitter 125 and the lens 130 to the analysis region 95. Irradiated with red blood cells 105 located within. In addition, the device for measuring the characteristics of the red blood cells may further include an analysis means such as a photodiode spectrometer (120).

Referring to FIG. 3A, the lower mirror 85, which is a multiple reflective layer, and the obtaining medium 90 are provided on the semiconductor substrate 150 to form a laser obtaining region. The upper mirror 80 and the insulating layer pattern 140, which are multiple dielectric layers formed on the substrate 150, form the analysis region 95 in which the red blood cells 105 are analyzed.

As shown schematically in FIG. 3B, a tube 155 through which red blood cells 105 passes is formed across the analysis region 95 and has a gate (at the end of the tube 155 and the analysis region 95). 160, the flow of red blood cells 105 is controlled. As such, the red blood cell 105 passing through the analysis region 95 is irradiated with the light ray 110, and the difference between the longitudinal and transverse wavelengths of the light scattered from the red blood cell 105 is analyzed to measure the degree of anemia.

As described above, conventionally, the shape and physical properties of erythrocytes were measured using a micropipette aspiration method or a filtration method. However, in the case of the micro pipette aspiration method, it is difficult to statistically analyze red blood cells in the whole blood based on a single cell because the amount of red blood cells is sucked into the micro pipette. Although it should have uniform dimensions, it is almost impossible to form the same size, shape and surface of the micropipette.

In addition, in the filtration method, a porous polycarbonate membrane is mainly used to measure the time for the entire sample to pass through the membrane, but in this case, only the average characteristic of the red blood cells can be measured. This is a disadvantage required. Moreover, it is difficult to grasp individual RBC characteristics of each red blood cell, and there is a problem in that various variables cannot be set.

In addition, in the case of a device for measuring the characteristics of the conventional red blood cells, since the characteristics of the red blood cells are analyzed by analyzing the difference between the wavelengths scattered from the red blood cells in the longitudinal and transverse directions, the scattered wavelengths are measured and compared to determine the characteristics. Many additional devices are required for this, and the process of obtaining the results is not only time-consuming but also very difficult to obtain accurate results.

Accordingly, an object of the present invention is to provide a device for diagnosing a disease and a method for diagnosing the disease, using physical properties of erythrocytes, which are sufficiently available for advanced diagnosis or early detection of a disease associated with deformability of erythrocytes such as cancer.

Another object of the present invention is to physically evaluate red blood cells, which can directly evaluate the effects of drugs administered to improve blood circulation disorder by controlling the deformity of blood and viscosity of blood in diseases such as atherosclerosis and blood circulation disorder. A disease diagnosis apparatus using the characteristics and a method of diagnosing the same are provided.

Another object of the present invention is to detect the physical properties of erythrocytes capable of diagnosing diseases related to blood by detecting changes in mechanical properties of erythrocytes according to changes in the viscosity of blood in the case of diseases related to blood, such as diabetes or malaria. It is to provide a disease diagnosis apparatus used and a diagnosis method thereof.

1 is a schematic diagram of a device capable of determining the shape of a conventional red blood cell.

2 is a schematic diagram of a device capable of measuring the properties of a conventional red blood cell.

3A is a cross-sectional view of the microcavity portion of the apparatus capable of measuring the characteristics of the red blood cells shown in FIG.

FIG. 3B is a plan view of the microcavity shown in FIG. 3A.

4 is a schematic perspective view of a disease diagnosis apparatus using physical properties of red blood cells according to an embodiment of the present invention.

5 is a cross-sectional view of a microchip applied to the apparatus for diagnosing diseases using physical properties of red blood cells according to the present invention.

6A and 6B are schematic diagrams for describing a process of measuring physical properties of red blood cells from an image acquired through an imaging unit.

7 and 8 are schematic perspective views and cross-sectional views illustrating a process of injecting a sample into a microchip for performing a method for diagnosing a disease according to an embodiment of the present invention, respectively.

9 and 10 are schematic perspective views and cross-sectional views illustrating a process of injecting a sample into a microchip for performing a method for diagnosing a disease according to another embodiment of the present invention, respectively.

11 is a schematic configuration diagram of a disease diagnosis apparatus using physical properties of red blood cells according to another embodiment of the present invention.

12A and 12B are schematic cross-sectional views illustrating a process of measuring physical properties of red blood cells using an image processing means according to the present invention.

13A and 13B are schematic diagrams for explaining a process of measuring physical properties of red blood cells through a signal generating means according to the present invention.

<Description of the code | symbol about the principal part of drawing>

200, 400: Disease diagnosis apparatus 210, 410: Microchip

220, 440: optical detection member 230, 420: fixed plate

240, 425: fixing member 245, 430: light generating member

250: frame grabber 255: imaging element

260: analyzer 295: sample injection member

297: Sample discharge | release member 320: Filter part

325, 330: first and second channel 335: inlet hole

340: discharge hole 345: inlet tube

350: discharge tube 370, 470: red blood cells

450: signal processing member 460: signal acquisition member

According to a preferred embodiment of the present invention to achieve the above object of the present invention, a fixing plate on which a microchip formed with a filter portion through which red blood cells pass is disposed, an optical detection member disposed on the microchip, the optical detection Provided is a disease diagnosis apparatus using physical properties of red blood cells, including an imaging device connected to a member and an analysis device connected to the imaging device.

The optical detection member is composed of a microscope or lens array to enlarge the light passing through the filter portion of the microchip, and the photographing element is configured of a CCD camera for capturing a continuous image to obtain an image of the filter portion. The analyzing apparatus further includes a frame grabber that analyzes an image obtained by the photographing element, outputs a result, and is connected to the photographing element to convert an image signal into a digital signal.

Preferably, the disease diagnosis apparatus further includes a light generating member disposed under the fixing plate to irradiate laser light or ultraviolet light having a wavelength of about 350 to 450 nm to the microchip. In addition, the disease diagnosis apparatus further includes a sample injection member composed of a chickenpox or a syringe pump and a sample discharge member composed of a syringe or a suction pump in order to inject and discharge blood of an examinee into the microchip.

In addition, according to another embodiment of the present invention, in order to achieve the above object of the present invention, a fixing plate to which a microchip formed with a filter portion through which red blood cells pass is fixed, a light generating member disposed below the fixing plate, the micro An apparatus for diagnosing a disease using physical properties of erythrocytes is provided that includes an optical detection member disposed on the chip, a signal acquisition member connected to the optical detection member, and a signal processing member connected to the signal acquisition member.

The signal acquisition member having a PMT or photodiode generates an electrical signal by detecting a change in brightness of light as the red blood cells pass through the microchip, and the signal processing member including an oscilloscope or a computer may be configured to generate the electrical signal. Analyze the signal and output the result.

In addition, according to a preferred embodiment of the present invention in order to achieve the above object of the present invention, the step of placing a microchip formed with a filter portion through which red blood cells pass on the fixing plate, the filter unit in the lower portion of the fixing plate Disposing a light generating member for irradiating light, disposing an optical detecting member for enlarging the light transmitted through the filter unit of the microchip, and connecting a photographing element to the optical detecting member to filter the filter; There is provided a disease diagnosis method using physical properties of red blood cells, including obtaining a negative image and analyzing an image obtained by connecting an analysis device to the imaging device and outputting a result. In this case, the length, shape and deformability of the red blood cells passing through the filter part are determined by obtaining an image of the red blood cells in one detection area of the filter part, and the speed of the red blood cells passing through the filter part is detected in two places of the filter part. The distance between the two areas in the area and the image of the erythrocytes are obtained and determined.

In addition, according to another preferred embodiment of the present invention in order to achieve the above object of the present invention, the step of placing a microchip formed with a filter portion through which red blood cells pass on the fixing plate, the lower portion of the microchip in the fixing plate Disposing a light generating member for focusing light on a filter unit; disposing an optical detecting member on the upper portion of the microchip to enlarge light passing through the filter unit of the microchip; and placing a signal obtaining member on the optical detecting member. Connecting to sense a change in brightness of light due to the passage of red blood cells to generate an electrical signal; and connecting a signal processing member to the signal acquisition member to analyze the electrical signal and outputting a result. A diagnostic method for diagnosing disease using the physical characteristics of the present invention is provided. At this time, the length, shape and deformability of the erythrocytes passing through the filter portion focus the light generated from the light generating means into one detection region of the filter portion, and the erythrocytes pass through the detection region and the length of the erythrocytes. And determining that the light is partially obscured according to the shape and the degree of deformation, and the velocity of the red blood cells passing through the filter unit focuses the light generated from the light generating unit in the first and second detection regions of the filter unit. And measure that the red blood cells partially pass the light in the first and second detection regions according to the red blood cells while passing through the first and second regions, and time for moving the red blood cells to the first and second regions. It is determined by measuring.

As described above, according to the disease diagnosis apparatus using the physical characteristics of the red blood cells according to the present invention, the speed, the length, the length of the red blood cells contained in the blood of the subject passing through the microchip by the optical detecting member and the imaging member or the signal acquisition member Physical properties of red blood cells, such as deformability and shape, can be easily measured. Therefore, it is possible to determine whether a disease such as cancer, malaria, or diabetes occurs from various results regarding the physical characteristics of the red blood cells measured by the disease diagnosis apparatus.

Although the apparatus for diagnosing diseases using the physical characteristics of red blood cells according to the present invention is insufficient to be used for accurate diagnosis and evaluation of cancer, it is possible to test for cancer using blood alone, and thus it is not related to the site of cancer. Compared with the test, early diagnosis of cancer is possible regardless of the site, so it can be used effectively for the early diagnosis and early detection of cancer.

In addition, the disease diagnosis apparatus using the physical characteristics of the red blood cells according to the present invention can be utilized to evaluate the effect of the drug to be administered when there is a disease such as atherosclerosis or blood circulation disorder. Diseases such as atherosclerosis or blood circulation disorders are difficult to determine with this device because of problems related to blood vessels themselves. However, in the case of drugs that improve blood circulation disorders by controlling the deformability of red blood cells and the viscosity of blood, the direct effects of the present invention It can be determined as a disease diagnosis device using the physical characteristics of the red blood cells.

Furthermore, in the case of diseases such as diabetes or malaria associated with blood, the mechanical properties of red blood cells change as the viscosity of the blood changes, so even when the disease diagnosis apparatus using the physical properties of the red blood cells is applied, While this may be difficult to distinguish from, this problem can be improved by scaling up the entire database and improving the system, and rather, it can be developed to measure various diseases at once.

Hereinafter, an apparatus for diagnosing a disease and a method for diagnosing the disease using physical properties of red blood cells according to preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. It doesn't happen.

Figure 4 shows the overall configuration of the disease diagnosis apparatus using the physical characteristics of the red blood cells according to an embodiment of the present invention.

Referring to FIG. 4, the disease diagnosis apparatus 200 using the physical characteristics of the red blood cells according to the present embodiment includes a fixing plate 230 on which the microchip 210 is fixed and a microchip mounted on the fixing plate 230. An optical detecting member 220 disposed above the 210 and configured as a microscope or a lens array, an image capturing device 255 connected to the optical detecting member 220, and a frame grabber connected to the capturing device 255. grabber) 250, and an analysis device 260 for analyzing the obtained image and outputting the result. In addition, the disease diagnosis apparatus 200 using the physical characteristics of the red blood cells is disposed under the fixing plate 230 to irradiate light to the microchip 210 disposed on the fixing plate 230 to obtain an accurate image. The light generating member 245 is included.

The fixing plate 230 serves to fix the microchip 210 disposed thereon so as not to move during the measurement process, and between the optical detection member 220 and the light generating member 245 while the measurement is performed. In the microchip 210 is moved upward or downward. When the fixing plate 230 is made of a light transmitting material such as, for example, a light transmitting plastic, the microchip 210 is irradiated from the light generating member 245 disposed under the fixing plate 230. Properties such as the speed, shape and deformability of red blood cells passing through the inside of the can be more easily measured.

A fixing member 240 such as a clip or a fixing ring is formed at one side of the fixing plate 230 to fix the microchip 210 to the fixing plate 230 while measuring physical properties of red blood cells located inside the microchip 210. ). However, the fixing member 240 may not be installed when the microchip 210 is attached to the fixing plate 230 or when the receiving groove in which the microchip 210 is accommodated is provided in the fixing plate 230. .

5 is a cross-sectional view of the microchip applied to the apparatus for diagnosing diseases using physical properties of red blood cells according to the present invention.

As shown in FIG. 5, the microchip 210 includes a first layer 310, which is an upper layer, and a second layer 315, which is a lower layer, each made of a light-transmitting material. The first layer 310 and the second layer 315 are made of light-transmitting plastic and glass, such as polydiallylmethylamine (PDMA) or polymethylmethacrylate (PMMA), and thus are separated from the light generating member 245 through the microchip 210. The irradiated light may penetrate the microchip 210.

A filter unit 320 including a plurality of thin film patterns spaced apart from each other at predetermined intervals is formed on the second layer 315, and a first portion that functions as an input unit and an output unit, respectively, is formed at the periphery of the filter unit 320. And a channel portion including the second channels 325 and 330. Each of the thin film patterns constituting the filter unit 320 may be a metal such as titanium, aluminum, tantalum, tungsten or platinum, a metal oxide such as titanium oxide, aluminum oxide, tantalum oxide, tungsten oxide or platinum oxide, silicon nitride, or TEOS ( It consists of an insulator such as Tetraethylorthosilicate to Low Temperature Oxide (LTO), and the first and second channels 325 and 330 are each made of photoresist. In this case, the thin film patterns are formed to have a thickness of about 1 to 4 μm and a width of about 3 to 7 μm, and the photoresist constituting the first and second channels 325 and 330 is about 25 to 100 μm. It is formed to a thick height.

An inflow hole 335 exposing a portion of the first channel 325 is formed from one upper portion of the first layer 310, and the second channel 330 is formed from the other upper portion of the first layer 310. A discharge hole 340 exposing a part is formed. When a sample such as blood of the examinee is injected into the microchip 210 having the above-described configuration from a sample injection means (not shown), the injected sample passes through the first channel 325 and passes through the filter unit 320. The optical detection member 220 enlarges the red blood cells passing through the filter unit 320. In this case, when the sample is introduced by using the chicken pox by connecting the first connection tube 345 to the inlet hole 335 or by using a syringe pump, the blood may be subjected to various characteristics of the blood while passing through the filter unit 320. Measurement is performed, and the measured blood is discharged through the discharge hole 340. In addition, when the second connection tube 350 is connected to the discharge hole 340 and the sample is sucked by using a pump or a syringe, the sample passes through the filter part 320 through the inlet hole 335 and the measurement is performed. After discharged to the discharge hole (340). That is, both the first and second connection tubes 345 and 350 may be connected to one of the inlet hole 335 and the outlet hole 340, respectively. For example, when the sample is introduced into the microchip 210, when the first connection tube 345 is inserted into the inlet hole 335 and a sample injection means such as a chicken pox or a syringe pump is used, the sample is introduced into the inlet hole. After entering through 335, the sample is naturally discharged through the discharge hole 340 after the characteristics of the sample are measured while passing through the filter unit 320. On the contrary, when the second connection tube 350 is connected to the discharge hole 340 and the sample is sucked out using a pump or a syringe, the sample is introduced through the inlet hole 335 and passes through the filter unit 320. It is measured and then discharged through the discharge hole 340.

Referring back to FIG. 4, the light generating member 245 is preferably a light source that emits a laser or ultraviolet (UV) light. However, light may be irradiated to the microchip 210 using a general lamp. The light generating member 245 irradiates the microchip 210 through which red blood cells pass through laser light or ultraviolet rays having a wavelength of about 350 to 450 nm, preferably about 400 nm.

In the present invention, as described above, since light having a wavelength of about 350 to 450 nm, preferably about 400 nm is used, human red blood cells typically absorb light having a wavelength of about 400 nm. Because there is a characteristic. That is, when irradiating red blood cells passing through the microchip 210 with a wavelength of about 400 nm from the light generating member 245, the red blood cells absorb light and appear black, and thus pass through the microchip 210. You can see red blood cells much better. Accordingly, accurate images of erythrocytes and erythrocytes can be acquired to accurately measure physical characteristics such as erythrocyte shape, movement speed, and length.

The optical detection member 220, which is a microscope or lens array, enlarges the filter unit 320 of the microchip 210 through which red blood cells pass, and captures an image enlarged by the optical detection member 220. (255) is taken. Preferably, the imaging device 255 is composed of a CCD (Charge Coupled Device) camera, the CCD camera captures the shape of the red blood cells passing through the filter unit 320 of the microchip 210, a plurality of Obtain an image. In this case, the CCD camera continuously photographs and compares a series of consecutive photographs taken by the CCD camera, thereby extracting the shape, length, and speed of the red blood cells to determine the degree of deformability of the red blood cells.

This will be described with reference to the drawings.

6A and 6B are schematic diagrams for describing a process of measuring physical properties of red blood cells from an image acquired through the imaging device 255. FIG. 6A is an image photograph of red blood cells 370 passing through the filter part 320 of the microchip 210 at any time t, and FIG. 6B shows red blood cells passing through the filter part 320 after a time Δt. 370 is a video picture. 6A and 6B, D denotes the length of the red blood cells 370, and S denotes the distance that the red blood cells 370 traveled for a time Δt.

6A and 6B, the length D of the red blood cells 370 and the degree of deformability of the red blood cells can be directly measured from either an image obtained at any time t or an image obtained at time t + Δt. . That is, the speed at which the red blood cells 370 pass through the filter unit 320 measures the distance S moved for a time equal to Δt, and when the moving distance S of the red blood cells 370 is divided by the time Δt, the unit time The movement speed of the sugar red blood cells 370 is obtained. Therefore, physical properties such as deformability, length, and speed of the red blood cells 370 may be specified to diagnose the presence or absence of a disease therefrom.

The frame grabber 250 connected to the photographing element 255 converts an image signal obtained through the photographing element 255 such as a CCD camera into a digital signal so that an analysis device 260 such as a computer can read it. It is preferably installed in the device 260. Based on the converted digital signal, the analysis device 260 analyzes the image to accurately determine the length, speed, and degree of deformability of the red blood cells 370 passing through the filter unit 320. In the present exemplary embodiment, the optical detection member 220, the imaging device 255, the frame grabber 250, and the analysis device 260 are described and illustrated separately, but the members are collectively integrated to acquire, process, and analyze a series of images. It could be implemented as a system that does this.

Hereinafter, a method of diagnosing a disease using a disease diagnosis apparatus using physical properties of red blood cells according to an embodiment of the present invention will be described.

7 and 8 illustrate schematic perspective views and cross-sectional views illustrating a process of injecting a sample into a microchip to perform a method for diagnosing a disease according to an embodiment of the present invention. 7 and 8, the same reference numerals are used for the same members as in FIG. 8, the small arrow shows the flow of the sample supplied from the sample supply means and injected into the microchip.

4, 7 and 8, the microchip 210 is fixed on the fixing plate 230 by using the fixing member 240, and then the head of the microchip 210 in the inlet hole 335. Alternatively, the connection tube 350 of the sample injection member 295 such as a syringe pump or the like is inserted. Subsequently, the sample is first injected into the sample injection member 295, and then a sample is supplied from the sample injection member 295 to the microchip 210 while applying a constant pressure to the sample injection member 295. According to this method, since the sample can be injected into the microchip 210 at a constant speed, there is an advantage that the speed of the red blood cells passing through the microchip 210 can be kept constant.

Samples such as red blood cells of the test subject injected from the sample injection member 295 pass through the inside of the microchip 210 along a small arrow direction. That is, the sample is first formed in the first layer 310 is introduced into the microchip 210 through the inlet hole 335 connected to the sample injection means 290 and the inlet tube 345, the first channel 325 Pass through the filter unit 320. The sample passing through the filter unit 320 is discharged to the outside through the discharge hole 340 through the discharge tube inserted into the second channel 330 and the first layer 310 again.

The red blood cells included in the sample pass through the filter unit 320 of the microchip 210 and the length and the degree of deformability of the red blood cells by the optical detecting member 220 and the imaging device 255 at the point indicated by the arrow 'A'. Is taken as a direct image is measured by the analysis device 260, the moving speed of the red blood cells is measured by the analysis device 260 is analyzed by a series of images taken by the imaging device 255. At this time, the light generating member 245 irradiates the red blood cells with light having a wavelength of about 350 to 450 nm, and preferably about 400 nm to the filter portion 320 of the microchip 210 to form the shape and length of the red blood cells. And precisely measure images for speed. This measurement process has already been described with reference to FIGS. 6A and 6B.

In the present invention, when a test subject suffers from a disease such as cancer, red blood cells are hardened, so deformability of red blood cells is reduced, so that more time is required to pass through the filter unit 320. However, since the red blood cells are slightly different in size, when the presence or absence of cancer is determined only by the speed at which the red blood cells pass through the filter unit 320 of the microchip 210, the probability of misdiagnosis is high. In the above) the red blood cells are measured together with the length of the deformation in the filter unit 320 to compare the two to increase the accuracy of the cancer diagnosis. For example, the analysis device 260 shows the measured values with the red blood cell moving speed as the X-axis and the red blood cell deformation as the Y-axis, thereby showing the data of the red blood cells of a normal subject and a patient with cancer. Comparing the data of the red blood cells of the data of the normal red blood cells and the red blood cells of the person with cancer occupy different areas on the XY plane, it is possible to determine the presence of cancer.

On the other hand, when the filter unit 320 of the microchip 210 is designed to be destroyed while the red blood cells pass through, the red blood cells of a person who has cancer pass through the filter unit 320 and are not immediately destroyed, but with a slight time delay. In this case, the presence or absence of cancer can be determined even by measuring the delay time.

In addition, in a patient with a disease such as malaria, malaria bacteria penetrate the inside of red blood cells, thereby making red blood cells harder, so some red blood cells with malaria cannot pass through the filter part 320 of the microchip 210. The presence or absence of such diseases can be determined.

9 and 10 are schematic perspective views and cross-sectional views illustrating a process of injecting a sample into a microchip to perform a method for diagnosing a disease according to another embodiment of the present invention, respectively. 9 and 10, the same reference numerals are used for the same members as in FIG. In addition, in FIG. 10, the small arrow shows the flow of the sample supplied from the sample supply means and inject | poured into a microchip.

4, 9, and 10, the microchip 210 is mounted on the fixing plate 230 using the fixing member 240 installed on the fixing plate 230, and then the microchip 210 of the microchip 210 is mounted. Drop the blood containing the red blood cells 370 of the person being examined in the inlet hole 335 by one drop. Subsequently, the syringe is connected to the discharge hole 345 of the microchip 210 and positioned in the inlet hole 335 of the microchip 210 using a sample discharge member 297 such as a syringe or a suction pump. The blood 370 is introduced into the microchip 210. Subsequently, the size, shape, and movement speed of the red blood cells 370 passing through the filter portion 320 of the microchip 210 using the light generating member 245, the optical detection member 220, and the imaging device 255. Measure physical properties such as At this time, since the process of analyzing the physical characteristics of the red blood cells 370 is the same as described above, a description thereof will be omitted. In addition, the sample discharge member 297 may use a small suction apparatus, and may also allow the blood 370 to pass through the microchip 210 manually.

As described above, the sample discharge member such as a suction pump through the connection tube 340 to the discharge hole 345 of the microchip 210 in order to smoothly flow the blood 370 of the tester into the microchip 210. 297 is installed. In addition, the red blood cells 370 may be introduced into the microchip 210 by simply connecting a syringe instead of the suction pump. In this method, it is easy to slow or fast adjust the speed of the red blood cells 370 passing through the microchip 210, and the amount of sample required to measure the physical characteristics of the red blood cells 370 is minute. Even in this case, it is possible to measure. In addition, since the sample can be flowed into the microchip 210 even without a separate additional device, the configuration of the device is simplified.

When diagnosing a disease using the microchip according to the present invention described above, undiluted general blood may be directly used, and when used in a hospital, only red blood cells may be separated to measure physical properties of the microchip. In addition, the extracted red blood cells may be diluted and used at a specific concentration such as about 1: 1 or 1: 2, and the red blood cells may be separated on the microchip according to the present invention, or the red blood cells may be separated using a concentration adjusting element. Can be used to adjust the concentration. Moreover, the microchip may be processed so that other microchannels having a structure similar to the microchip according to the present invention are added to the microchip in order to adjust the concentration of the sample.

Hereinafter, an apparatus for diagnosing a disease and a method of diagnosing the disease using physical properties of red blood cells according to another embodiment of the present invention will be described in detail with reference to the accompanying drawings.

Figure 11 shows the overall configuration of the disease diagnosis apparatus using the physical characteristics of the red blood cells in accordance with another embodiment of the present invention.

Referring to FIG. 11, the disease diagnosis apparatus 400 according to the present exemplary embodiment includes a fixing plate 420 on which the microchip 410 is fixed, and a light generating member 430 installed below the fixing plate 420. An optical detection member 440 disposed on the microchip 410, a signal acquisition member 460 connected to the optical detection member 440, and a signal processing member 450 connected to the signal acquisition member 460. ).

Since the configuration of the microchip 410 according to the present embodiment is the same as that shown in FIG. 5, a description thereof will be omitted.

The light generating member 430 emits laser light, ultraviolet light, or other lamp light, and is disposed under the fixing plate 420 and fixed to the fixing plate 420 to obtain an accurate image. ) Is focused on the filter section. In this case, the fixing member 425 may be installed in the fixing plate 420 to secure the fixing stability when the microchip 410 moves. Also in this embodiment, the light generating member 430 is about 350-450 nm, preferably in order to acquire accurate images of erythrocytes and erythrocytes and accurately measure physical properties such as erythrocyte shape, movement speed and length. Is irradiated to the microchip 410 through which red blood cells pass a laser or ultraviolet light having a wavelength of about 400 nm.

When the fixing plate 420 is made of a light-transmitting material, the speed of red blood cells passing through the inside of the microchip 410 by light emitted from the light generating member 430 disposed below the fixing plate 420. Since the characteristics such as shape and deformability can be more easily measured, the fixing plate 420 is preferably made of a material having excellent light transmittance.

A fixing member 425 such as a clip or a fixing ring is formed at one side of the fixing plate 420 to stably hold the microchip 410 to the fixing plate 420 while measuring physical properties of red blood cells in the microchip 410. Can be fixed with In addition, the optical detection member 440 is configured as a lens array or a microscope as described above, and serves to enlarge the filter unit of the microchip 410 through which red blood cells pass, and enlarged by the optical detection member 440. Image is transmitted to the signal acquisition member 460. According to this embodiment. The signal acquisition member 460 is composed of a photo-multiplier tube (PMT) or a photo diode.

12A and 12B show schematic cross-sectional views for explaining a process of measuring physical properties of erythrocytes using the apparatus according to the present embodiment. In FIGS. 12A and 12B, the arrow indicates the advancing direction of the red blood cells 470 passing through the filter part of the microchip 410.

12A and 12B, the signal acquiring member 460 according to the present exemplary embodiment includes a PMT or a photodiode, and a filter of the microchip 410 through which red blood cells enlarged by the optical detecting member 440 passes. Change in brightness of negative light is detected. Unlike the CCD camera described above, the PMT or the photodiode is an apparatus that displays an electrical signal according to a change in brightness of light.

When the microchip 410 is installed on the fixing plate 420 and the light is emitted while the focus of the light generating member 430 is set to the filter portion of the microchip 410, the red blood cells 470 may remove the portion. As the light passes, the light is obscured, and thus the brightness of the light changes. Accordingly, the PMT or the photodiode generates an electrical signal. The signal processing member 450 connected to the signal acquisition member 460 analyzes the electrical signal generated from the PMT or the photodiode and measures the time that the red blood cell 470 passes, thereby extracting the characteristic value of the red blood cell 470. do. In this case, the signal processing member 450 is composed of an oscilloscope, a computer, or other equivalent circuit device for displaying an electrical signal generated from the signal acquisition member 460.

13A and 13B are schematic diagrams for describing a process of measuring physical properties of red blood cells through the signal acquisition member 460. FIG. 13A is a view for explaining a process of measuring characteristics such as length, shape, and deformability of red blood cells, and FIG. 13B is a view for explaining a process for measuring a moving speed of red blood cells.

Referring to FIG. 13A, the light irradiated from the light generating member 430 is focused to a predetermined area C of the filter portion of the microchip 410. When the red blood cells 470 pass through the filter unit and a part of the luminous flux is obstructed according to the length or degree of deformation of the red blood cells, the signal acquiring member 460 detects this and generates an electrical signal, which is transmitted to the signal processing member 450. By converting the signal into a readable signal, the length of the red blood cells 470, the degree of deformation and the like can be measured.

Referring to FIG. 13B, in order to measure the speed of the red blood cells 370, when the red blood cells 470 pass through the filter part of the microchip 410, two first and second detection areas D1 and D2 are located on the filter part. ). First, the signal acquiring member 460 detects a change in light according to the passage of the red blood cells 470 of the first detection region D1 at an arbitrary time t, and generates an electrical signal. Then, at a time t + Δt, the second detection region ( In D2), the signal acquisition member 460 detects a change in light according to passage of the red blood cells 470 to generate an electrical signal. Electrical signals generated in the first and second detection regions D1 and D2 are transmitted to the signal processing apparatus 450 to determine the moving speed of the red blood cells 470 by the signal processing apparatus 450. That is, the length and the deformability of the red blood cells 470 determine the length and the degree of deformability of the red blood cells 470 by measuring the change in the luminous flux of the light emitted from the light generating member 430 by the red blood cells 470. The speed at which the red blood cells 470 pass through the filter unit is measured by detecting a change in light as the red blood cells 470 pass through the designated first and second detection areas D1 and D2.

A method of diagnosing a disease using a disease diagnosis apparatus using physical properties of red blood cells according to another embodiment of the present invention excludes a process of detecting physical properties of red blood cells by a signal acquisition member and a signal processing member as described above. If the same as the case of the embodiment of the present invention described above it will be omitted.

In addition, the process of injecting a sample into the microchip to perform the method for diagnosing a disease according to the present embodiment is the same as in the above-described embodiment, so a detailed description thereof will be omitted.

According to the disease diagnosis apparatus using the physical characteristics of the red blood cells according to the present invention, such as the speed, length, deformability and shape of the red blood cells contained in the blood of the subject passing through the microchip by the optical detection member and the imaging device or the signal acquisition member Physical properties can be easily measured. Therefore, it is possible to determine whether there is a disease such as cancer, malaria, or diabetes from various results regarding the physical characteristics of the red blood cells measured by the disease diagnosis apparatus.

Although the apparatus for diagnosing diseases using the physical characteristics of red blood cells according to the present invention is insufficient to utilize for accurately diagnosing or evaluating a disease such as cancer, since it is possible to test cancer by blood alone and has no relation to the site of cancer Compared with conventional biopsies, early diagnosis of cancer is possible regardless of the site, so it can be used very effectively for the early diagnosis and early detection of cancer.

In addition, the disease diagnosis apparatus using the physical characteristics of the red blood cells according to the present invention can be utilized to evaluate the effect of the drug to be administered when there is a disease such as atherosclerosis or blood circulation disorder. Diseases such as atherosclerosis and blood circulation disorders are related to blood vessels, so they are difficult to determine with this device.However, in the case of drugs that improve blood circulation disorders by controlling the deformability of blood cells and the viscosity of blood, red blood cells have seen the direct effects. It can be determined as a disease diagnosis device using the physical properties of.

Furthermore, in the case of diseases such as diabetes or malaria, which are related to blood, the mechanical properties of red blood cells change because the viscosity of the blood changes, so even when applying a disease diagnosis apparatus using the physical properties of the red blood cells, Sometimes this can be difficult to distinguish, but this problem can be improved by scaling up the entire database and improving the system, rather than by developing multiple diseases at once.

As described above, although described with reference to preferred embodiments of the present invention, those skilled in the art will be variously modified and modified without departing from the spirit and scope of the invention described in the claims below. It will be appreciated that it can be changed.

Claims (26)

In the disease diagnosis apparatus using the physical characteristics of the red blood cells, A fixed plate on which a microchip having a filter portion through which red blood cells pass is disposed; Optical detection means arranged on the microchip to enlarge light passing through the filter portion of the microchip; Photographing means connected to the optical detecting means to obtain an image of the filter part; And An apparatus for diagnosing diseases using physical properties of red blood cells, characterized in that it comprises an analysis means for analyzing the image obtained by being connected to the photographing means and outputting the result. The method of claim 1, The apparatus for diagnosing diseases using physical properties of erythrocytes, further comprising a light generating means disposed under the fixing plate to irradiate light to the microchip. The method of claim 2, The light generating means is a disease diagnosis device using the physical characteristics of the red blood cells, characterized in that for emitting a laser light or ultraviolet light having a wavelength of 350 ~ 450nm. The method of claim 1, The optical detection means is a disease diagnosis device using the physical characteristics of the red blood cells, characterized in that consisting of a microscope or a lens array. The method of claim 1, The photographing means is a disease diagnosis device using the physical characteristics of the red blood cells, characterized in that consisting of a CCD camera for the continuous imaging of the image. The method of claim 1, The analyzing means further comprises a frame grabber connected to the photographing means and converting an image signal into a digital signal. The method of claim 1, Device for diagnosing disease using the physical characteristics of the red blood cells, characterized in that the microchip further comprises a sample injection means for injecting and discharging the blood of the subject. The method of claim 7, wherein The sample injection means is a disease diagnosis device using the physical characteristics of the red blood cells, characterized in that it comprises a chickenpox or syringe pump. The method of claim 1, Device for diagnosing disease using the physical characteristics of the red blood cells, characterized in that the microchip further comprises a sample discharge means for injecting and discharging the blood of the subject. The method of claim 9, The sample discharge means is a disease diagnosis device using the physical characteristics of the red blood cells, characterized in that it comprises a syringe or a suction pump. In the disease diagnosis apparatus using the physical characteristics of the red blood cells, A fixing plate to which the microchip on which the filter unit through which red blood cells pass is fixed; Light generating means disposed under the fixing plate to irradiate light to the filter portion of the microchip; Optical detection means arranged on the microchip to enlarge light passing through the filter portion of the microchip; Signal acquisition means connected to the optical detection means for detecting a change in brightness of light due to the passage of red blood cells and generating an electrical signal; And And a signal processing unit connected to the signal obtaining unit and analyzing the electrical signal to output a result. The method of claim 11, The light generating means is a disease diagnosis device using the physical characteristics of the red blood cells, characterized in that for emitting a laser light or ultraviolet light having a wavelength of 350 ~ 450nm. The method of claim 11, The optical detection means is a disease diagnosis device using the physical characteristics of the red blood cells, characterized in that it comprises a lens array or a microscope. The method of claim 11, The signal acquisition means is a disease diagnosis apparatus using the physical characteristics of the red blood cells, characterized in that it comprises a PMT or a photodiode. The method of claim 14, The signal processing means is a disease diagnosis device using the physical characteristics of the red blood cells, characterized in that it comprises an oscilloscope or a computer. The method of claim 11, Device for diagnosing disease using the physical characteristics of the red blood cells, characterized in that the microchip further comprises a sample injection means or sample discharge means for injecting the blood of the subject. The method of claim 16, The sample injection means comprises a chickenpox or syringe pump, and the sample discharge means comprises a syringe or a suction pump disease diagnosis apparatus using the physical characteristics of the red blood cells. In a method for diagnosing a disease using physical properties of red blood cells, Positioning a microchip having a filter portion through which red blood cells pass on a fixing plate; Disposing a light generating means for irradiating light to the filter portion below the fixing plate; Disposing optical detection means on the upper part of the microchip to enlarge light transmitted through the filter part of the microchip; Connecting an image pickup means to the optical detection means to obtain an image of the filter part; And And analyzing the image obtained by connecting the analyzing means to the photographing means, and outputting a result of the disease. The method of claim 18, The light generating means is a disease diagnosis method using the physical characteristics of the red blood cells, characterized in that for emitting a laser light or ultraviolet light having a wavelength of 400nm. The method of claim 18, The step of connecting the analyzing means further comprises the step of connecting the frame grabber for converting an image signal into a digital signal to the photographing means disease diagnosis method using the physical characteristics of the red blood cells. The method of claim 20, The length, shape and deformability of the red blood cells passing through the filter part are determined by obtaining an image of the red blood cells in one detection area of the filter part. The method of claim 18, The rate of erythrocytes passing through the filter unit is determined by obtaining the distance between the two regions and the image of the erythrocytes in the two detection regions of the filter unit, characterized in that the physical characteristics of the red blood cells. The method of claim 22, And connecting the sample injecting means or the sample discharging means to inject and discharge the blood of the test subject into the microchip. In a method for diagnosing a disease using physical properties of red blood cells, Positioning a microchip having a filter portion through which red blood cells pass on a fixing plate; Disposing a light generating means for focusing light on a filter part of the microchip below the fixing plate; Disposing optical detection means on the upper part of the microchip to enlarge light passing through the filter part of the microchip; Connecting a signal acquisition means to the optical detection means to detect a change in brightness of light due to passage of the red blood cells to generate an electrical signal; And And connecting the signal processing means to the signal acquiring means to analyze the electrical signal and to output a result. The method of claim 24, The length, shape and deformability of the erythrocytes passing through the filter portion focus the light generated from the light generating means into one detection region of the filter portion, and the erythrocyte length, shape and A method for diagnosing a disease using physical properties of red blood cells, characterized in that the light is partially obscured according to the degree of deformation. The method of claim 24, The speed of the red blood cells passing through the filter unit focuses the light generated from the light generating means in the first and second detection regions of the filter unit, and the red blood cells are set according to the red blood cells while passing through the first and second regions. A method for diagnosing a disease using physical properties of red blood cells, characterized in that the light is partially obscured in the first and second detection areas, and measured by measuring the time that the red blood cells travel through the first and second areas.