WO2014142559A1 - Apparatus and method for detecting and counting rare cells in blood - Google Patents

Abstract

The present invention relates to an apparatus and a method for detecting and counting rare cells in blood. The apparatus according to the present invention comprises a blood sample collection tube, a measurement kit, a detector, and the like, wherein a complex material in which a target antibody and a tracer are combined, and blood are dispersed and mixed in the blood sample collection tube, blood is injected and individual cells are captured at respective micro-trap structures in the measurement kit, and the number of the micro-trap structures, which emit light by the occurrence of an antigen-antibody reaction and the adhering of tracers to the surface of cells, can be counted in the detector. According to the present invention, it is possible to easily and conveniently detect and count the number of rare cells through a normal blood test at clinical practice and test multiple samples.

Description

Apparatus and method for detecting and counting rare cells in blood

The present invention relates to a device and method for detecting and counting rare cells present in blood. More specifically, the present invention relates to a complex material in which blood rare cells such as circulating vascular endothelial cells (CEC) are combined with a simple target antibody and a label, and An apparatus and method for detecting and counting rare cells using a microgap plate.

Although the population of the blood is very small compared to known blood cells such as red blood cells, white blood cells, and platelets, there are very few cells that can play an important role in the spread of disease or can be used as biomarkers. It is called (rare cells).

Representative rare cells include CTC (Circulating Tumor Cell), and one or two of billions of blood cells are known to exist. In the case of a large number of CTCs, a blood test can diagnose a very definitive clue that can be determined to have cancer, and various blood test methods have been developed.

Recently, as clinical papers and related technologies have been reported that circulating endothelial cells (CECs) can be used as biomarkers for early diagnosis of cardiovascular diseases such as heart failure, CTC-like or related technologies are rapidly developing. It is a trend.

For example, according to the "Characterization of circulating endothelial cells in acute myocardial infarction" published in Science Translational Medicine in March 2012, the cardiovascular group was found to have about five times more CEC than the healthy group. If the number increases above a certain number, it is reported that early diagnosis of a disease such as a heart attack may occur within two weeks.

Although the characteristics of cells such as CTC or CEC are present in the blood, but their quantity is very small, it is very difficult to separate or detect them by general technology, but both core diseases of human death cause such as cancer and cardiovascular disease are involved. It is a very important rare cell in that it is a rare cell. Detection technology can be difficult, but for the patient, providing only a small amount of blood (7.5 ml) allows for cancer screening and cardiovascular disease testing, which is a very innovative and market-waiting technology.

Currently known CTC or CEC detection methods are methods for detecting rare cells using antigen-antibody responses with very high selectivity. That is, by coating an antibody capable of attaching to an antigen of a specific rare cell on the surface of the magnetic nanoparticle and stirring it with a blood sample, the nanoparticle is attached to a specific rare cell in the blood. After strong fixation using magnetic force, the remaining cells can be washed to isolate specific rare cells. The isolated rare cells can be identified through a microscope using a staining technique for verification, and individual cell numbers can be counted.

However, these conventional methods require separate processes such as pretreatment and posttreatment, and there is an inconvenience in that such a process takes a very long time. In addition, it is very difficult to use in the clinical field due to the necessity of skilled inspectors to deal with such a process. Therefore, there is a need for a quantitative and objective measurement device and method for a simpler and faster test and a medical practitioner for easy real-time use in a clinical setting.

Accordingly, an object of the present invention is to solve the problems of the prior art as described above, and to provide an apparatus and a method capable of quickly and accurately detecting and counting the rare cells existing in a small amount from a certain amount of blood.

In order to achieve the above object, the present invention collects a blood sample, a sample collection unit for receiving a complex material combined with a target antibody and a label; A measurement kit connected to the sample collection unit for injecting a mixture of the blood sample and the composite material, and individually trapping blood cells; And a detector for detecting and counting the rare cells having generated the antigen-antibody reaction with the target antibody among the blood cells captured in the measurement kit.

In the present invention, the target antibody may specifically recognize and bind to rare cells in the blood.

Labeling material in the present invention is a fluorescent material; Quantum dots; Micro size beads in which quantum dots are integrated; It may include one or more selected from metal nanoparticles.

In the present invention, the blood sample collected in the sample collection unit may be stirred with the dispersion of the composite material.

In the present invention, the measurement kit includes two plates having a micro-sized gap, thereby allowing blood samples to be moved by capillary action.

In the present invention, the plate may be manufactured in a rigid plate-like structure or a flexible film-like structure.

In the present invention, one end of the measurement kit is formed with an injection hole into which a blood sample is injected, and the other end may be provided with a capillary tube formed with an opening through which air passes.

In the present invention, the opening of the capillary may be connected to a flow driving mechanism to inject and flow the blood sample into the measurement kit.

At least one of the two plates in the present invention may be provided with an uneven portion so that blood cells are individually captured.

In the present invention, a stepped stepped step is formed in the concave-convex portion to capture small cells at the bottom, and large cells can be captured at the top.

In the present invention, the detector includes a light source capable of exciting one or more of fluorescent materials, quantum dots, and micro-sized beads in which quantum dots are integrated; An image sensor for measuring excited fluorescence; By providing a fluorescence filter for passing light of a specific wavelength band, it may be a fluorescence scanning device for detecting the position and number of blood cells that emit light.

In the present invention, the light source may include at least one selected from a mercury lamp, a laser diode (LD), and a laser light emitting diode (LED).

In the present invention, the image sensor may include at least one selected from a photo diode array, a charge coupled device (CCD) array, and a complementary metal oxide semiconductor (CMOS) array.

In the present invention, a zoom lens may be mounted on the image sensor to acquire an optically enlarged image.

In the present invention, the light source and the image sensor may be installed in a form in which they are continuously coupled.

In the present invention, the detector is one selected from an optical device, a micro-sized prism array, and a nano-sized grating array, which can cause surface plasmon resonance of metal nanoparticles. By providing the above, it can be a surface plasmon resonance imaging scanning apparatus which detects the position and number of blood cells in which surface plasmon resonance occurs.

In the present invention, the surface plasmon resonance imaging scanning apparatus includes a light source for injecting parallel light into the measurement kit; And a camera for measuring light reflected from the measurement kit.

In the present invention, the measurement kit may be integrated with a micro-sized prism array or a nano-sized grating array.

The device according to the invention comprises a moving rail connected to the detector and guiding the movement of the detector for scanning of the measurement kit; A mobile collection device for sampling after detection; And a two-axis moving rail connected to the mobile collecting device and guiding the movement of the mobile collecting device.

In addition, the present invention using the apparatus described above, the step of collecting a blood sample with a sample collection unit; Stirring the collected blood sample to be mixed with a complex material combined with a target antibody and a label; Injecting a mixture of the blood sample and the complex into a measurement kit to capture blood cells individually; And detecting and counting the rare cells in which the antigen-antibody reaction has occurred with the target antibody among the blood cells captured in the measurement kit by using a detector.

According to the present invention, a blood sample tube which can be used for one-time use is mixed with the target antibody and the labeling substance, and the blood is mixed, and then the number of micro traps that emits light while being injected into the measurement kit plate is quickly and easily and conveniently. In addition to detecting and counting rare cells, it is also possible for a healthcare practitioner to objectify and standardize detection and counting with simple operation. In particular, the time required for one test is short, so a large amount of test is possible.

1 schematically shows an apparatus and method for detecting and counting rare cells in blood according to the present invention.

2 is a plan view of a measurement kit according to the invention.

3 is a cross-sectional view showing a mechanism in which cells are captured in a measurement kit having a concave-convex structure according to the present invention.

Figure 4 is a cross-sectional view showing a mechanism for trapping cells in the measurement kit having a step-shaped concave-convex structure in accordance with the present invention.

5 is a plan view of a device for detecting and counting rare cells in the blood according to the present invention.

6 is a front view of a device for detecting and counting rare cells in the blood according to the present invention.

7 is a block diagram of a fluorescence scanning device among detectors according to the present invention.

8 is a block diagram of a surface plasmon resonance imaging scanning apparatus of the detector according to the present invention.

9 is a perspective view showing that a micro-sized prism array or a nano-sized grating array used in the surface plasmon resonance imaging method according to the present invention is integrated with a measurement kit.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

Figure 1 schematically shows the apparatus and method for detecting and counting blood rare cells according to the present invention, the apparatus for detecting and counting blood rare cells 100 according to the present invention is a rare cell diagnostic system, the sample collection unit 10 ), A measurement kit 20, a detector 30, and the like.

The sample collection unit 10 collects a blood sample and also serves to receive the composite material 11. That is where the blood sample is loaded.

The sample collection unit 10 may be formed in a tube shape and may have a stopper. Preferably, the sample collection unit 10 may have a structure of a blood collection tube of 7.5 ml volume, which is generally widely used.

The blood sample may include a plurality of blood cells (B), and the blood cells (B) may be largely divided into small cells (S) such as red blood cells and large cells (L) such as white blood cells and rare cells.

The composite material 11 may be in the form of combining the labeling material 12 and the target antibody 13. The labeling substance 12 and the target antibody 13 may be chemically or physically coupled through a linker. The linker is not particularly limited, and conventional linkers may be used. For example, peptide linkers, hydrazine linkers, disulfide linkers, and the like may be used.

The composite material 11 may be provided in the form of a solution containing the composite material 11, preferably in the form of a dispersion in which the composite material 11 is dispersed in a dispersion medium. For example, the composite material dispersion may include a dispersant such as the composite material 11, a dispersion medium such as water, a dispersant such as a surfactant, and the like.

As the labeling material 12, one or more of 1) fluorescent material, 2) quantum dots, 3) micro-sized beads in which quantum dots are integrated, and 4) metal nanoparticles may be selected and used.

The fluorescent material is not particularly limited and conventional fluorescent materials can be used, for example, fluorescein, isothiocyanate, rhodamine, phycoerythrin, and phycoerythine. Phycocyanin, allophycocyanin, phthaldehyde, fluorescamine and the like can be used. The size of the fluorescent material is not particularly limited, and may be, for example, 1 to 900 nm, preferably 10 to 500 nm, more preferably 30 to 300 nm.

The quantum dot may include a central particle and a ligand bound to the surface of the central particle. Examples of the material forming the core particles include II-VI compound semiconductor nanocrystals such as CdS, CdSe, CdTe, ZnS, ZnSe, ZnTe, HgS, HgSe, HgTe, III- such as GaN, GaP, GaAs, InP, InAs. And group V compound semiconductor nanocrystals or mixtures thereof. The central particle may have a core / shell structure, and each of the core and shell of the central particle may include the compounds exemplified above. Exemplary compounds may be included in the core or shell each alone or in combination of two or more. For example, the central particle may have a CdSe-ZnS (core / shell) structure having a core comprising CdSe and a shell comprising ZnS. The ligand can prevent the central particles adjacent to each other from easily agglomerated and quenched with each other. In addition, the ligand may bind to the central particle so that the central particle has hydrophobicity. As an example of a ligand, an amine compound, a carboxylic acid compound, etc. which have a C6-C30 alkyl group are mentioned. As another example of a ligand, the amine compound, carboxylic acid compound, etc. which have a C6-C30 alkenyl group are mentioned. The size of the quantum dots is not particularly limited and may be, for example, 1 to 900 nm, preferably 10 to 500 nm, more preferably 30 to 300 nm.

The micro-sized beads in which the quantum dots are integrated may be a capsule structure surrounding a quantum dot integrated body in which a plurality of quantum dots are integrated. Capsules can be prepared using gelatin, cellulosic compounds, or the like. The size of the microbeads is not particularly limited and may be, for example, 0.1 to 900 μm, preferably 0.5 to 500 μm, more preferably 1 to 100 μm.

The metal constituting the metal nanoparticles is not particularly limited, and all conventional metals may be used. For example, gold, silver, copper, iron, platinum, tungsten, titanium, tin, nickel, chromium, cobalt, zinc, iridium Etc. can be used. Moreover, a metal oxide, a metal salt, etc. can be used, or an alloy can also be used. The size of the metal nanoparticles is not particularly limited, and may be, for example, 1 to 900 nm, preferably 10 to 500 nm, more preferably 30 to 300 nm.

As the target antibody 13, an antibody capable of specifically recognizing and binding rare cells in the blood such as CTC and CEC may be used. For example, Anti-EpCAM antibodies can be used.

After a certain amount of blood sample is collected in the sample collection unit 10, the blood sample may be stirred to mix with the composite dispersion therein. The stirring may be performed by a manual method of shaking by hand, or may be performed by an automatic method using a stirrer or the like. The mixing order of the blood sample and the composite material dispersion is not particularly limited, and the blood sample may be first put into the sample collection unit 10 or the composite material dispersion may be put first. If a specific blood rare cell is present during the stirring process, a large amount of the complex material 11 to which the labeling substance 12 and the target antibody 13 are bound may be attached through an antigen-antibody reaction around the cell membrane.

In the lower right of Figure 1, a scanning image for cell counting (Cell Counting) is illustrated, where a black portion of the plurality of grids represents a hybrid rare cells (Hybridized RC) to which the composite material is bound.

Figure 2 is a plan view of the measurement kit according to the present invention, the measurement kit 20 is provided with a flat plate 21, 22, inlet 23, capillary 24, uneven structure (25, 26, 27, 28), etc. can do.

The measurement kit 20 serves to individually trap cells in the blood. The measurement kit 20 may be connected to the sample collection unit 10 to inject a mixture of the blood sample and the composite material 11. To this end, an injection hole 23 into which a blood sample is injected may be formed at one end of the measurement kit 20. The injection port 23 may be configured, for example, in the form of a needle, or may include a needle structure. For example, after the sample collecting unit 10 is upside down, the plug of the sample collecting unit 10 is plugged into the needle-shaped injection hole 23 or the needle structure of the injection hole 23, and the sample collecting unit 10 As the measurement kit 20 is connected, a mixture of the blood sample and the composite material stirred in the sample collection unit 10 may be injected into the measurement kit 20.

The measurement kit 20 may be composed of a microfluidic sheet for cell spreading. Preferably, the measurement kit 20 includes two plates 21, 22 with micro-sized gaps, thereby allowing blood samples to be moved by capillary action. However, the present invention is not limited thereto, and the measurement kit 20 may be composed of one flat plate or three or more flat plates. The gap between the two plates 21, 22 may be for example 1 to 500 μm, preferably 5 to 100 μm, more preferably 10 to 50 μm.

The plates 21 and 22 can be made of rigid plate-like structures or flexible film-like structures. The flat plates 21 and 22 may be made of metal, plastic, glass, ceramic, or the like, and may be preferably transparent to facilitate detection. The thicknesses of the flat plates 21, 22 can be, for example, from 1 μm to 10 mm, preferably from 10 μm to 5 mm, more preferably from 50 μm to 3 mm.

When the two plates 21 and 22 are arranged in parallel at regular intervals, the edge portion between the two plates 21 and 22, that is, the left and right sides and the front and rear surfaces between the two plates 21 and 22 may be opened. This open portion may be sealed using a sealing material or the like. It is also possible to fabricate the measurement kit 20 as an integral structure in the form of a very thin cube without openings. Of course, the injection port 23 and the capillary tube 24 may be formed separately.

Capillary tube 24 may serve to allow a mixture of blood sample and composite to flow well between two plates 21, 22 of measurement kit 20. To this end, the capillary tube 24 may be formed at the other end of the measurement kit 20, for example opposite the inlet 23. The capillary tube 24 may have an opening through which air is communicated with the outside. The opening may for example be formed at the end of the capillary tube 24. The diameter of the capillary tube 24 may be, for example, 0.01 to 100 μm, preferably 0.1 to 50 μm, more preferably 0.5 to 20 μm, such that only air can pass therethrough.

A flow drive mechanism may be connected to the distal opening of the capillary 24 to inject the blood sample into the measurement kit 20 at a suitable rate and also to flow the blood sample inside the measurement kit 20. The flow drive mechanism may comprise a vacuum pump, for example. For example, if a vacuum is applied to the inside of the measurement kit 20 by using a vacuum pump and then stopped, the blood sample flow may be advanced and then reversed (backflow), so that the overall blood sample may be spread evenly. In addition, by adjusting the pressure of the vacuum pump, it is possible to control the injection rate and the flow rate of the blood sample.

3 is a cross-sectional view illustrating a mechanism in which cells are captured in a measurement kit having a concave-convex structure according to the present invention, in which at least one of the two plates 21 and 22 is trapped so that blood cells B are individually trapped. Parts 25 and 26 may be provided. Although the uneven portions 25 and 26 are formed in the lower plate 22 in the drawing, the present invention is not limited thereto, and the uneven portions 25 and 26 may be formed in the upper plate 21, and both flat plates 21 and 22 may be formed. ) May be formed on both.

Concave- convex portions 25 and 26 are composed of concave portion 25 and convex portion 26, and preferably, may be formed in a microstructure of micro units. The number of the uneven parts 25 and 26 is not particularly limited and depends on the size of the measurement kit 20, preferably 10 or more, more preferably 100 or more. As many as 10,000, 100,000, or more than 1 million are possible.

The width, length, and height (depth) of the uneven parts 25 and 26 may be each independently, for example, 0.1 to 200 µm, preferably 0.5 to 70 µm, and more preferably 1 to 30 µm.

In the figure, the movement direction of the blood sample is indicated by an arrow, and it can be confirmed that blood cells B are captured by the lumbar portion 25.

On the other hand, the flow driving mechanism allows the flow of the blood sample between the two plates 21 and 22 to be forward or backward to flow backward so that the blood cells B are individually captured in the traps of the uneven portions 25 and 26. By repeatedly inducing one cell (B) can be trapped in one uneven portion (25, 26).

4 is a cross-sectional view showing a mechanism in which the cells are captured in the measurement kit having a step-shaped concave-convex structure according to the present invention, the step-shaped step is formed in the concave-convex portions (25, 26, 27, 28) small cells (S ) Can be captured at the bottom, and large cells (L) can be captured at the top.

As shown in the figure, the stepped concave-convex structure may be a structure in which a small concave portion 27 is further formed below the bottom of the large concave portion 25. In addition, both upper side surfaces of the large convex portion 26 may be cut to form a small convex portion 28, or a small convex portion 28 may protrude from both lower sides of the large convex portion 26.

As such, the uneven parts 25, 26, 27, and 28 are formed in a step shape so that a step is formed, and as shown in the drawing, small cells S, such as red blood cells, are captured in the small recesses 27 that are lower. The large cells L, such as leukocytes and rare cells, can be captured in the large lumbar portion 25 directly above.

In addition, the uneven parts 25, 26, 27, and 28 may optimally form the depth (height), the width (width), and the like so that the captured cells S and L do not escape by the external flow. Since the size of the red blood cells is about 1 to 8 μm, and the size of the white blood cells is about 12 to 25 μm, the width and depth of the small recesses 27 can be configured to about 1 to 10 μm, and the width of the large recesses 25 is The depth can be configured to about 11 to 30 ㎛. As such, in the measurement kit 20, a blood sample may be injected so that individual cells B may be captured in each micro trap structure.

On the other hand, the measurement kit 20 to check the number of cells that emit light or surface plasmon resonance phenomenon in response to the composite material 11 among the cells captured in the uneven portion (25, 26, 27, 28) May be mounted on a pedestal or the like of the device 100.

5 is a plan view of a blood rare cell detection and counting device according to the present invention, Figure 6 is a front view, the blood rare cell detection and counting device 100 is a sample collector 10, the measurement kit 20, In addition to the detector 30, a moving rail 34, a movable collecting device 40, a biaxial moving rail 41, and the like may be provided.

The detector 30 detects and counts the rare cells in which the antigen-antibody reaction has occurred with the target antibody 13 among the blood cells B captured by the measurement kit 20. The detector 30 may detect the location and number of cells in the blood by an optical method. The detector 30 may include, for example, an ultra-precision CCD sensor array, and scan the measurement kit 20 to inspect the position and number of cells emitting light. The detector 30 may be configured as a fluorescence scanning device or a surface plasmon resonance imaging scanning device according to the type of the labeling material 12, which will be described later.

The moving rail 34 serves to guide the movement of the detector 30 for the scanning of the measurement kit 20. To this end, one end of the moving rail 34 may be mounted on a pedestal of the apparatus 100, and the other end may be connected to the detector 30. The detector 30 may move in the up and down direction of the drawing along the moving rail 34. The detector 30 may be provided with wheels, rollers, etc. to be movable along the moving rail 34. The moving rail 34 is fixed and may not move. The movement of the detector 30 may be manual movement by hand or automatic movement using a motor or the like.

The mobile collection device 40 serves to take a sample after detection. The collecting device 40 may be configured in the form of, for example, a syringe. In the case of using a syringe, a sample may be taken by drilling the upper plate 21 of the measurement kit 20. The collecting device 40 can also be moved manually or automatically using a motor or the like.

The two-axis moving rail 41 serves to guide the movement of the movable collecting device 40. As illustrated in the drawings, the biaxial moving rail 41 may be composed of a vertical rail and a horizontal rail, and the collecting device 40 may be mounted at an intersection point of the two rails. The movable collecting device 40 may move along the biaxial moving rail 41 in the up and down direction and the left and right direction of the drawing. The two-axis moving rail 41 may be installed to move on its own without being fixed to a pedestal of the apparatus 100, and may move together with the movable collecting device 40.

When the scanning of the detector 30 is completed, the detection position of the rare cells may be addressed so that the mobile collection device 40 may move to the target position along the biaxial movement rail 41 to collect the rare cells. The collected sample can be used as a sample for secondary precision analysis.

7 is a block diagram of a fluorescence scanning device among detectors according to the present invention, wherein the labeling material 12 is at least one of 1) fluorescent material, 2) quantum dots, and 3) micro-sized beads in which the quantum dots are integrated. In this case, the detector 30 may be configured as a fluorescence scanning device.

The fluorescence scanning device 30 includes a light source 31 capable of exciting one or more kinds of fluorescent materials, quantum dots, and micro-sized beads in which quantum dots are integrated; An image sensor 32 capable of measuring excited fluorescence; By providing the fluorescent filter 33 which can pass light of a specific wavelength band, it is possible to detect the position and number of blood cells which emit light.

The light source 31 may include at least one selected from a mercury lamp, a laser diode (LD), and a laser light emitting diode (LED).

The image sensor 32 may include at least one selected from a photo diode array, a charge coupled device (CCD) array, and a complementary metal oxide semiconductor (CMOS) array.

The image sensor 32 may be equipped with a zoom lens to acquire an optically enlarged image.

The light source 31 and the image sensor 32 may be installed in a form in which they are continuously coupled.

The wavelength band of light that the fluorescent filter 33 can pass may vary depending on the type of fluorescent material or quantum dots used.

With respect to the measurement kit 20 having completed the detection process, it is possible to check the appearance of the cell at the point of light emission using an optical device such as a microscope.

8 is a block diagram of the surface plasmon resonance imaging scanning apparatus of the detector according to the present invention. When the labeling material 12 is a metal nanoparticle, the detector 30 may be configured as the surface plasmon resonance imaging scanning apparatus 50. have.

The surface plasmon resonance imaging scanning device 50 is an optical device capable of generating surface plasmon resonance of metal nanoparticles, a micro-sized prism array, and a nano-sized grating array. By providing at least 1 sort (s) chosen from the inside, the position and number of the blood cells which surface plasmon resonance phenomenon generate | occur | produce can be detected.

The surface plasmon resonance imaging scanning apparatus 50 includes a light source 51 for injecting parallel light 53 into the measurement kit 20, as shown in the figure; And a camera 52 measuring the reflected light 54 reflected from the measurement kit 20.

As such, the surface plasmon resonance imaging scan device 50 may inject parallel light 53 into the measurement kit 20 and measure the reflected light 54 with the camera 52. When the metal nanoparticles are attached to the rare cells in the blood in a large amount, and the light comes in at a specific incident angle at the location, the light is absorbed only at the location, and the amount of reflected light is different from other parts of the measurement kit 20. Can be detected. For example, light of 400 to 900 nm wavelength may be incident at an angle of 10 to 80 degrees. Preferably, light of 600 to 800 nm wavelength may be incident at an angle of 30 to 70 degrees.

9 is a perspective view showing that the micro-sized prism array or the nano-sized grating array used in the surface plasmon resonance imaging method according to the present invention is integrated with the measurement kit, and the measurement kit 20 generates the surface plasmon resonance phenomenon. In order to create a condition that may be, it may be integrated with a micro sized prism array 60 or a nano sized grating array.

The prism array 60 or the grating array is integrated with the outer surface of the measurement kit 20. Specifically, the prism array 60 or the grating array may be bonded to the outer surface of the upper plate 21 or the lower plate 22 by bonding, welding, injection, extrusion, or the like. Can be integrated.

The shape of the prism array 60 or the grating array, as shown in the figure, it is preferable that the cross section is a triangle, but can be changed to a circle, oval, polygon, etc. as necessary.

The size of the prism array 60 may be, for example, 1 μm to 1 mm, preferably 10 to 500 μm, more preferably 100 to 200 μm. The size of the grating array can be for example 1 nm to 10 μm, preferably 10 nm to 3 μm, more preferably 100 nm to 1 μm.

[Description of the code]

10: sample collector

11: composite material

12: Labeling Substance

13: target antibody

20: measuring kit

21, 22: reputation

23: injection hole

24: capillary

25, 26, 27, 28: uneven portion

30: detector (fluorescence scanning device)

31: light source

32: image sensor

33: fluorescent filter

34: moving rail

40: mobile collection device

41: 2-axis movable rail

50: surface plasmon resonance imaging scanning device

51: light source

52: camera

53: parallel light

54: reflected light

60: prism array (grating array)

100: rare cell detection / counting device (rare cell diagnosis system)

Claims (20)

1. A sample collecting unit for collecting a blood sample and containing a complex material in which a target antibody and a labeling substance are combined;

   A measurement kit connected to the sample collection unit for injecting a mixture of the blood sample and the composite material, and individually trapping blood cells; And

   A device for detecting and counting blood rare cells, comprising a detector for detecting and counting rare cells in which an antigen-antibody reaction has occurred, among blood cells captured in a measurement kit.
2. The method of claim 1,

   Target antibody is a detection and counting device for rare cells in the blood, characterized in that to specifically recognize and bind the rare cells in the blood.
3. The method of claim 1,

   The labeling substance is a fluorescent substance; Quantum dots; Micro size beads in which quantum dots are integrated; Device for detecting and counting rare cells in the blood, characterized in that it comprises one or more selected from metal nanoparticles.
4. The method of claim 1,

   The blood sample collected in the sample collection unit is a device for detecting and counting rare cells in the blood, characterized in that the agitated with a dispersion of the composite material.
5. The method of claim 1,

   The measuring kit is provided with two plates having a micro-sized gap, whereby the blood sample can be moved by capillary action.
6. The method of claim 5,

   A device for detecting and counting rare cells in blood, wherein the plate may be made of a rigid plate-like structure or a flexible film-like structure.
7. The method of claim 5,

   One end of the measurement kit is formed with an injection port for injecting a blood sample, the other end of the blood rare cell detection and counting device, characterized in that the capillary formed with an opening through which the air passes.
8. The method of claim 7, wherein

   A flow driving mechanism is connected to the opening of the capillary tube to inject and flow a blood sample into the measurement kit.
9. The method of claim 5,

   At least one of the two plates is provided with a concave-convex portion so that blood cells are captured separately, the blood rare cells detection and counting apparatus.
10. The method of claim 9,

    An uneven portion is formed in a stepped step, small cells are captured in the lower portion, large cells are detected and counting devices, characterized in that the large cells are captured in the upper portion.
11. The method of claim 3,

    The detector includes a light source capable of exciting one or more of fluorescent materials, quantum dots, and micro-sized beads in which the quantum dots are integrated; An image sensor for measuring excited fluorescence; A fluorescence scanning device for detecting the position and number of blood cells that emit light by providing a fluorescence filter for passing light of a specific wavelength band, wherein the detection and counting device for rare blood cells in blood.
12. The method of claim 11,

    The light source includes at least one selected from a mercury lamp, a laser diode (LD), and a laser light emitting diode (LED).
13. The method of claim 11,

    An image sensor includes at least one selected from a photo diode array, a charge coupled device (CCD) array, and a complementary metal oxide semiconductor (CMOS) array.
14. The method of claim 11,

    The image sensor is equipped with a zoom lens to detect and count the rare cells in the blood, characterized in that to obtain an optically enlarged image.
15. The method of claim 11,

    And a light source and an image sensor are installed in a continuous coupling form.
16. The method of claim 3,

    The detector is provided with at least one selected from optical devices, micro-sized prismatic arrays and nano-sized grating arrays, which can generate surface plasmon resonance of metal nanoparticles. And a surface plasmon resonance imaging scanning device for detecting the position and number of blood cells in which surface plasmon resonance occurs.
17. The method of claim 16,

    The surface plasmon resonance imaging scanning apparatus includes a light source for injecting parallel light into the measurement kit; And a camera for measuring light reflected from the measurement kit.
18. The method of claim 16,

    And a measurement kit is integrated with a micro-sized prism array or a nano-sized grating array.
19. The method of claim 1,

    A moving rail coupled to the detector and guiding movement of the detector for scanning of the measurement kit;

    A mobile collection device for sampling after detection; And

    A device for detecting and counting rare cells in blood, coupled to a mobile collection device, further comprising a biaxial moving rail for guiding the movement of the mobile collection device.
20. Using the apparatus of claim 1,

    Collecting a blood sample with a sample collector;

    Stirring the collected blood sample to be mixed with a complex material combined with a target antibody and a label;

    Injecting a mixture of the blood sample and the complex into a measurement kit to capture blood cells individually; And

    A method for detecting and counting rare cells in blood, the method comprising detecting and counting rare cells having generated an antigen-antibody reaction with a target antibody among blood cells captured in a measurement kit.

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