KR101060957B1 - Biomaterial Detection Device and Biomaterial Measurement System - Google Patents

Abstract

The present invention relates to a biomaterial detection apparatus and a biomaterial measurement system, and an aspect of the present invention provides a body portion having a detection region therein, and one surface and the other surface of the body portion to allow fluid to pass through the detection region. It provides a biological material detection device comprising a spherical microbead is disposed in the fluid injection portion and the fluid discharge portion and the detection region of the body portion formed on the surface, and the material capable of capturing the biological material is coated on the surface.

According to the present invention, it is possible to obtain a biomaterial detection device having a biochip structure and a measurement system capable of easily and precisely measuring the biomaterial captured therein so as to have excellent specific cell detection performance and excellent mass productivity.

Cell, circulating tumor cell, CTC, blood, detection, instrumentation

Description

Biomaterial Detection Device and Biomaterial Measurement System {DEVICE AND MEASURING SYSTEM FOR DETECTING BIO MOLECULE}

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a biological material detection device and a biological material measurement system, and more particularly, to a biological material detection device and a biological material measurement system having a biochip structure to have excellent specific cell detection performance and excellent mass productivity.

Recently, the demand for methods or apparatuses for detecting target molecules (DNA, RNA, etc.), specific proteins, and Rare Cells in the blood is increasing in the biomedical germ. For example, in Circulating Tumor Cells (CTCs), there is a small amount of one billionth the specific gravity in the blood, and the presence of CTCs can be precisely determined for the prediagnosis of diseases such as cancer. There is a need for a technique that can be detected. However, in the case of rare cells such as CTC, it is very difficult to detect and count in an accurate and fast time because of the extremely low concentration in the blood, and therefore, various methods for detecting rare cells have been studied in the art. .

One object of the present invention is to provide a biomaterial detection apparatus having a biochip structure to have excellent specific cell detection performance and excellent mass productivity. Another object of the present invention is to provide a biomaterial measurement system capable of precisely measuring the biomaterial detection device.

In order to achieve the above object, one aspect of the present invention,

A body portion having a detection region therein, and a fluid injection portion and a fluid discharge portion formed on one surface and the other surface of the body portion so as to allow fluid to pass through the detection region, and a detection region of the body portion, Provided is a biological material detection device comprising a spherical microbead coated on a surface of a trapping material.

In one embodiment of the present invention, the plurality of microbeads may be provided with different diameters. In this case, the detection area may be divided into a plurality of areas formed by gathering ones having the same diameter among the plurality of microbeads. Furthermore, the plurality of areas are provided closer to the fluid injection unit. The diameter of the microbeads may be large.

In one embodiment of the invention, the material capable of capturing the biological material is preferably provided with one material selected from the group consisting of antigen, antibody and conductive polymer.

In one embodiment of the present invention, the body portion may be flat.

In one embodiment of the present invention, the plurality of microbeads may be arranged in a matrix structure forming a row and a column as a plurality.

In an embodiment of the present invention, the microbeads may be arranged to be in contact with each other as a plurality of adjacent ones.

In one embodiment of the present invention, the micro bead may be optically exposed to the outside of the body portion.

In one embodiment of the present invention, the micro bead is preferably made of a light transmissive material.

In one embodiment of the present invention, the micro bead may be disposed in the groove formed in the body portion.

In one embodiment of the present invention, the diameter of the microbead is preferably 5㎛ or more, more preferably, the diameter of the microbead may be 5 ~ 200㎛.

In one embodiment of the present invention, the fluid injection portion and the fluid discharge portion may be formed on the surface facing each other respectively.

Another aspect of the invention,

A body portion having a detection region therein, a fluid injection portion and a fluid discharge portion formed on one surface and the other surface of the body portion so as to allow fluid to pass through the detection region, and a detection region of the body portion, It provides a biological material detection system comprising a biological material detection device having a spherical microbead coated on the surface of the captureable material and an optical unit for measuring the biological material captured by the biological detection device. .

In one embodiment of the present invention, the biological material is preferably combined with a fluorescent material.

In an embodiment of the present disclosure, the optical device may include a CCD camera and a lens disposed on at least one of the upper and lower portions of the detection area. In this case, an image of the detection area photographed by the CCD camera may be divided into a grid and stored in the image processing apparatus. The apparatus may further include a stage accommodating the biological material detection device and movable in one direction and another direction perpendicular thereto.

In one embodiment of the present invention, the optical device may be an optical pickup device having a laser diode and a light receiving unit disposed on at least one of the upper and lower portions of the detection area.

According to the present invention, it is possible to obtain a biomaterial detection device having a biochip structure and a measurement system capable of easily and precisely measuring the biomaterial captured therein so as to have excellent specific cell detection performance and excellent mass productivity.

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

However, embodiments of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below. In addition, embodiments of the present invention are provided to more completely describe the present invention to those skilled in the art. Accordingly, the shape and size of elements in the drawings may be exaggerated for clarity, and the elements denoted by the same reference numerals in the drawings are the same elements.

1 is a perspective view showing a biological material detection apparatus according to an embodiment of the present invention, Figures 2 and 3 are respectively a top plan view and a perspective view showing an aspect in which the biological material is detected in the microbead in Figure 1. First, referring to FIG. 1, the biological material detecting apparatus 100 according to the present embodiment includes a body portion 101, a micro-bead 102, a fluid injection portion, and a fluid discharge portion 103, 104. It is a structure having a structure, and after the specific material of the biological material contained in the fluid, such as blood injected from the fluid injection unit 103 is captured by the microbead 102 constituting the detection area, the remaining fluid is the fluid discharge unit 104 To get out.

The body portion 101 has a detection area therein for detecting a biological material as blood passes therethrough, and may have a structure in which the upper body 101a and the lower body 101b are coupled as shown in FIG. 1. . The detection region may include a plurality of microbeads 102 disposed in grooves formed in the upper body 101a and the lower body 101b. On the contrary, after forming a groove in only one of the upper body 101a and the lower body 101b, a structure in which a transparent cover is covered thereon may also be employed. In this case, the microbead 102, which is a biological material capture means, may be optically exposed to the outside to measure the biological material by an optical device, and although not clearly illustrated in FIG. 1, the microbead 102 may be the body It may have a structure that can be measured from the lower portion of the portion (101). That is, the lower body 101b has a structure having a central hole and a transparent lower cover formed thereon, or the lower body 101b itself may be made of a transparent material, and thus, the upper body and the lower part may be doubled. Measurements can be made to improve measurement speed. In addition, the present invention is not limited thereto, but the body portion 101, as shown in Fig. 1, has a flat-chip structure that is easy to handle at the time of measurement in the form of a bio-chip (Bio-Chip).

The fluid injector and the fluid outlet 103 and 104 may have a flow path structure for functioning as an inlet and outlet passage of a fluid such as blood in the biomaterial detection apparatus 100. To this end, the fluid inlet and the fluid outlet 103, 104 is preferably formed on one surface and the other surface facing the body portion 101, according to the embodiment the fluid inlet and the fluid outlet ( The positions of 103 and 104 can be changed as appropriate.

The microbead 102 has a spherical shape, and serves as an obstacle for blocking the flow of a fluid such as blood. In the present embodiment, the microbeads 102 may be separately manufactured to form a detection area in a manner that is filled in the body portion 101, and the inside of the body portion 101 may be partially etched. It's not the way to get rid of obstacles. As such, the mass productivity of the biochip may be improved by not undergoing the etching process.

As shown in FIGS. 2 and 3, when blood passes through the detection region, specific biomaterials (C), such as target molecules (DNA, RNA, etc.), specific proteins, may appear on the surface of the microbeads 102. Intravital cells, such as CTCs, can be captured. To this end, the surface of the microbead 102 is coated with a suitable material to enable antigen / antibody binding. Specifically, the surface of the microbead 102 may be coated by selecting an appropriate material, for example, an antigen or an antibody, depending on the type of biological material to be detected, or may be coated with a conductive polymer in addition to these materials.

If the biomaterial to be detected is CTC, the surface of the microbead 102 may be coated with a biomaterial or chemical that specifically reacts with the CTC as a capture means. As a representative example, anti-EpCAM that specifically forms antigen-antibody binding with EpCAM (Epithelial Cell Adhesion Molecule) on the surface of CTC, or an aptamer having a DNA or RNA structure equivalent to or greater than that may be used. . In addition to EpCAM, biochemicals or chemicals that can specifically bind to specific proteins expressed on the surface of the CTC and distinguished from other nucleated cells (such as white blood cells) are available. For efficient coating of such biomaterials or chemicals, functional groups such as amine groups (-NH2), carboxyl groups (-COOH), thiol groups (-SH), etc. may be positioned on the surface of the microbeads 102 at regular intervals. Such functional groups form strong covalent bonds with biomaterials such as antibodies and aptamers such as DNA, such as amide bonds, disulfide bonds, etc. It is attached. In this case, the detection efficiency can be improved by adjusting the interval of the functional group.

As shown in FIG. 1, the microbeads 102 have a matrix structure in rows and columns in the plate-shaped body portion 101, and may be arranged in contact with each other. By such an arrangement structure, a large number of microbeads 102 are provided in the detection area, thereby improving detection power. In this case, even if adjacent spheres are in contact with each other, as shown in FIG. 3, a sufficiently large area is secured in the upper and lower portions of the areas in contact with each other to allow the biomaterial C to pass therethrough.

In addition, in the case of a sphere, since the area of contact with the fluid is larger than that of other structures such as cylinders and polygonal columns, the detection force of the biological material C may be improved by employing the microbead 102. In this case, the microbead 102 is made of a transparent material in order to visually or microscopically observe all the biological material (C) attached to the spherical surface while the microbead 102 is located in the body portion 101 as it is. desirable. Examples of transparent materials include materials such as polymers, silica, glass, and the like, which are considered to be suitable for coating antibodies and the like in addition to light transmission.

Another advantage of the microbead 102 having a spherical shape is that the likelihood of trapping the biomaterial by reducing shear stress can be increased. That is, in general, when the obstacle in contact with the flowing fluid has a spherical shape, the shear stress in the area in contact with the fluid is lower than in the case of a cylinder or polygonal column. This reduction in shear stress can reduce the likelihood that cells, such as CTC, already trapped in the microbeads 102 are dropped by the flow of fluid and rejoin the fluid, thereby detecting the desired biological material (C). You can expect an improvement.

On the other hand, the microbead 102 may be determined in various ways depending on the size of the biological material to be detected without particular limitation on its size. However, when the size of the microbeads 102 is small, a plurality of microbeads 102 may be provided in the detection area of the same size, the cell capture ability can be improved. However, if the diameter is too small, the size of the region through which blood can pass is also small, so that rare cells, such as relatively large CTCs, may not pass through, so that the diameter of the microbead 102 is about 5 It is preferable to determine in the range of 5 micrometers or more, More specifically, 5-200 micrometers.

4 is a perspective view showing a biological material detection apparatus according to another embodiment of the present invention. The biomaterial detection apparatus 200 according to the present embodiment includes a body portion 201, micro beads 202a and 202b, a fluid injection portion and a fluid discharge portion 203 and 204, and the body portion 201 is It is the same as the embodiment of FIG. 1 in that it can be divided into an upper body 201a and a lower body 201b. The present embodiment differs from the previous embodiment in that the diameters of the microbeads 202a and 202b can be varied. That is, as shown in Figure 4, the detection region is provided with a microbead (202a) having a relatively large diameter and a microbead (202b) having a smaller diameter than the microbeads (202a, 202b) having different diameters are gathered It can be divided into two areas. In this case, the size of the microbead 202a may be larger in an area closer to the fluid injecting part 203, and in the opposite manner. In addition, although not shown separately, it may be divided into three or more regions having microbeads of different diameters. As in the present embodiment, the microbeads 202a and 202b can be diversified in diameter and disposed in stages according to the flow of the fluid, thereby providing the advantage of filtering the biomaterials of various sizes in stages.

The biological material detection device having the above-described structure may be simply measured by the naked eye or by a microscope, but may be more precisely measured by the optical device. 5 and 6 each show a biomaterial measurement system that can be obtained in another aspect of the present invention. First, as shown in FIG. 5, the biomaterial measuring system 300 includes a biomaterial detecting apparatus 100, a stage 301, a CCD camera 302, an objective lens 303, and a processing device corresponding to a sample. It is a structure provided with 304 and the controller 305.

The biomaterial detection apparatus 100 has a structure according to the embodiment of FIG. 1, that is, a biochip, and corresponds to a state of a sample in which a predetermined biomaterial is captured by passing blood or the like. In this case, the biological material and the fluorescent material may be combined to improve the detection power, and the binding of the fluorescent material may use a method known in the art.

As illustrated in FIG. 5, the stage 301 accommodates the biological material detecting device 100 on an upper surface thereof and may move left and right and up and down. According to this movement, the area to be detected is divided into grids, and each is stored as one image, and the number of fluorescences of the stored images can be counted. This process is performed by the processing unit 304 connected to the CCD camera 302, the movement of the stage 301 may be controlled by the controller 305 connected to the processing unit 304. In this case, depending on the embodiment, the CCD camera 302 may be moved instead of the stage 301.

6, the biomaterial measuring system 400 according to another embodiment uses an optical pickup device. That is, the biomaterial measuring system 400 includes a biomaterial detecting apparatus 100, a light source 401 such as a laser diode, a collimating lens 402, a beam splitter 403, an objective lens 404, and a color screening filter ( The dichroic filter 405, the sensor lens 406, and the light receiving part 407 are comprised. As the light source 401 and the biomaterial detection apparatus 100 move properly, the desired amount of biomaterial may be determined by counting the number of fluorescent materials.

The present invention is not limited by the above-described embodiments and the accompanying drawings, but is intended to be limited only by the appended claims. It will be apparent to those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. something to do.

1 is a perspective view showing a biological material detection apparatus according to an embodiment of the present invention, Figures 2 and 3 are respectively a top plan view and a perspective view showing an aspect in which the biological material is detected in the microbead in Figure 1.

4 is a perspective view showing a biological material detection apparatus according to another embodiment of the present invention.

5 and 6 each show a biomaterial measurement system that can be obtained in another aspect of the present invention.

<Description of the symbols for the main parts of the drawings>

101, 201: body parts 102, 202: micro beads

103, 203: fluid inlet 104, 204: fluid outlet

301: stage 302: CCD camera

303: objective lens 304: processing unit

305: controller 401: light source

402: collimation lens 403: beam splitter

404: objective lens 405: color filter

406: sensor lens 407: light receiving unit

Claims (20)

A body part having a detection area therein; A fluid injection part and a fluid discharge part respectively formed on one surface and the other surface of the body to allow fluid to pass through the detection area; And A spherical microbead disposed in the detection region of the body part and having a diameter of 5 μm or more and coated with a material capable of capturing a biological material on a surface thereof; Biological material detection apparatus comprising a. The method of claim 1, And a plurality of microbeads having different diameters. The method of claim 2, And said detection region is divided into a plurality of regions formed by gathering ones having the same diameter from each other among the plurality of microbeads. The method of claim 3, And the plurality of regions are closer to the fluid injecting portion, the larger the diameter of the microbeads provided therein. The method of claim 1, The biological material detection apparatus, characterized in that the material capable of capturing the biological material comprises one material selected from the group consisting of antigen, antibody and conductive polymer. The method of claim 1, The body material detecting device, characterized in that the body portion is flat. 7. The method according to claim 1 or 6, And a plurality of microbeads arranged in a matrix structure in rows and columns. 7. The method according to claim 1 or 6, And said plurality of microbeads are arranged so that adjacent ones are in contact with each other. The method of claim 1, The microbead is a biological material detection device, characterized in that optically exposed to the outside of the body portion. The method of claim 1, The microbead is a biological material detection device, characterized in that made of a light transmitting material. The method of claim 1, The microbead is a biological material detection device, characterized in that disposed in the groove formed in the body portion. delete The method of claim 1, The diameter of the microbead is a biological material detection device, characterized in that 5 ~ 200㎛. The method of claim 1, And the fluid injecting portion and the fluid ejecting portion are formed on surfaces facing each other. A body portion having a detection region therein, a fluid injection portion and a fluid discharge portion formed on one surface and the other surface of the body portion so as to allow fluid to pass through the detection region, and a detection region of the body portion, 5 μm A biological material detection device having the above diameter and including spherical microbeads coated on a surface of a material capable of capturing the biological material; And A measuring unit including an optical device for measuring a biological material captured by the biological detecting device; Biological material measurement system comprising a. The method of claim 15, The biomaterial is a biomaterial measurement system, characterized in that coupled to the fluorescent material. The method of claim 15, And the optical device has a CCD camera and a lens disposed on at least one of the upper and lower portions of the detection area. The method of claim 17, And the image of the detection area photographed by the CCD camera is divided into a lattice shape and stored in the associating apparatus. The method of claim 17, And a stage accommodating the biomaterial detection device and movable in one direction and the other direction perpendicular thereto. The method of claim 15, And the optical device is an optical pickup device having a laser diode and a light receiving unit disposed on at least one of the upper and lower portions of the detection area.

Images