KR20100091839A - Integrated bio-chip and method of fabricating the integrated bio-chip - Google Patents

Abstract

PURPOSE: An integrated bio-chip and a method for manufacturing the same are provided to simplify bio-material detection apparatus. CONSTITUTION: An integrated bio-chip comprises: a substrate(110) having a plurality of reaction zones(140) to which sample is attached; a sample reaction unit(100) having an excitation light absorption waveguide(120); and a sample detection unit(200) which is coupled to the sample reaction unit and to detect fluorescence released from the sample. The excitation light absorption waveguide comprises a color filter core which penetrates fluorescence emitted from the sample and absorbs excitation light. A method for manufacturing the integrated bio-chip comprises: a step of preparing the sample reaction unit having the plurality of excitation light absorption waveguide; a step of preparing the sample detection unit having a plurality of pixels; and a step of welding the sample reaction unit and sample detection unit.

Description

Integrated bio-chip and method of fabricating the integrated bio-chip

The present invention relates to an integrated biochip and a method of manufacturing the same, and more particularly, to an integrated biochip and a method of manufacturing the same is inserted into the excitation light absorption waveguide used for detecting a sample by a spectroscopic method.

Bio-chips generally have a structure in which very small cells of micro units are arranged in a matrix form on a substrate, and such cells have a bio-material such as nucleic acid or protein. The probe bio-material on the biochip's substrate functions as a biological receptor for the target biomaterial.

Biochips detect target organisms by using interactions between such organisms, such as hybridization of nucleic acids or antigen-antibody reactions. Such a biochip can be used as a tool for analyzing gene function, searching for disease-related genes, gene expression, protein distribution, and the like by detecting biomaterials such as nucleic acids or proteins having a specific base sequence.

Detection of interactions between biomass mainly uses fluorescence detection methods. This fluorescence detection method is a spectroscopic method of detecting a fluorescence image obtained by irradiating predetermined excitation light onto a fluorescent substance labeled on a biomaterial. The detection of the fluorescence image is made through a light detection device such as a CCD scanner or a CIS scanner.

Since the fluorescence obtained by irradiating predetermined excitation light to the fluorescent substance labeled on the biomaterial is very weak light compared to the excitation light to be irradiated, it is necessary to remove the excitation light. On the other hand, the current detection device using a commercially available biochip uses a complex and expensive scanner equipment as a photodetector, a more compact biochip and its manufacturing method is required.

Embodiments of the present invention provide an integrated biochip and a method for manufacturing the same, which can react a sample such as a biomaterial in an integrated chip and optically detect the sample.

According to an embodiment of the present invention, an integrated biochip includes a substrate provided with at least one reaction region to which a sample is attached, and a fluorescence which absorbs and emits excitation light provided in each of the at least one reaction region to excite the sample. A sample reaction unit having at least one excitation light absorption waveguide penetrating from the substrate surface provided with the at least one reaction region to a rear surface of the substrate surface; And a sample detection unit provided on a rear side of the substrate surface on which the plurality of reaction regions of the substrate are provided to detect fluorescence emitted from the sample and integrally coupled to the sample reaction unit.

The excitation light absorption waveguide may include a color filter core that transmits fluorescence emitted from a sample and absorbs excitation light that excites the sample. Here, the color filter core may have a refractive index higher than that of the substrate surrounding the color filter core.

The excitation light absorption waveguide may further include a cladding layer provided around the color filter core and having a refractive index lower than that of the color filter core. Such a cladding layer may be a single layer or multiple layers.

The cross section of the color filter core may be circular or polygonal.

The plurality of reaction regions may be provided with a micro lens for condensing fluorescence emitted from the sample.

It is provided in the plurality of reaction zones, it may further include a fluorescence antireflection film for transmitting the fluorescence emitted from the sample.

The sample detector may be a charge coupled device (CCD) or a CMOS image sensor (CIS).

Pixels of the sample detector may correspond one-to-one or one-to-many with the at least one reaction region.

The apparatus may further include a frame for mounting the sample reaction unit and the sample detection unit.

By protecting the sample reaction unit and the sample detection unit, it may further include a cover glass is an anti-reflection coating for the excitation light.

In accordance with another aspect of the present invention, there is provided a method of manufacturing an integrated biochip, the method comprising: providing a sample reaction unit having at least one excitation light absorption waveguide formed through a substrate; Providing a sample detector including at least one pixel; And bonding the sample reaction unit and the sample detection unit.

The preparing of the sample reaction part may include forming at least one through hole in the substrate; And filling a color filter material into each of the plurality of through holes to form a color filter core.

The preparing of the sample reaction part may further include forming a cladding layer on inner walls of the plurality of through holes before forming the color filter core.

The preparing of the sample reaction part may further include forming a microlens in a region in which an excitation light absorption waveguide of the substrate is formed on one surface of the substrate.

In the bonding of the sample reaction unit substrate and the sample detection unit substrate, the pixels of the sample detection unit may correspond one-to-one or one-to-many with the at least one excitation light absorption waveguide.

The sample reaction unit and the sample detection unit may be bonded in a wafer unit or an individual chip unit.

The method may further include surface treating a region where the excitation light absorption waveguide is formed so that the sample may be attached to one surface of the sample reaction unit.

According to the embodiments of the present invention, by integrating the sample reaction unit for detecting the fluorescence emitted from the sample reaction unit and the biological reaction reacted in a single chip, the integrated biochip and biomaterial detection apparatus using the same It can be made compact.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the examples exemplified below are not intended to limit the scope of the present invention, but are provided to fully explain the present invention to those skilled in the art. In the drawings, like reference numerals refer to like elements, and the size of each element in the drawings may be exaggerated for clarity and convenience of description.

1 schematically illustrates an integrated biochip in accordance with an embodiment of the present invention. 2 schematically shows the optical path of fluorescence emitted from a sample in an integrated biochip according to this embodiment.

Referring to FIG. 1, the integrated biochip of this embodiment has a structure in which the sample reaction unit 100 and the sample detection unit 200 are integrally coupled. Here, the expression 'integrated' may include a sample reaction unit 100 including a substrate on which a sample 190 corresponding to a conventional biochip may be attached, and a sample capable of detecting fluorescence emitted from the sample 190. This means that the detection unit 200 is integrated into an integrated chip.

The sample reaction unit 100 includes a substrate 110 having a plurality of reaction regions 140 to which the sample 190 is attached, and a plurality of excitation light absorption waveguides 120 formed through the substrate 110. do.

As the substrate 110, for example, a semiconductor substrate such as Si, GaAs, or InP may be used. In some cases, glass, a dielectric, a metal or a polymer may be used.

One surface of the substrate 110 is provided with a plurality of reaction regions 140 to which the sample 190 is attached. The plurality of reaction regions 140 are spaced apart from each other, and correspond to the plurality of excitation light absorption waveguides 120 one-to-one. For example, when used as a DNA chip, each of the reaction regions 140 is a minimum unit of a region to which a plurality of homologous nucleic acids are attached, and fluorescence obtained when the excitation light is illuminated on the DNA chip to detect a target nucleic acid. It is the minimum pixel of the image. The reaction region 140 may have a diameter of, for example, sub μm to several μm and may be arranged in two dimensions such as a matrix. In FIG. 1, the reaction region 140 may be formed in a circular shape or a polygonal shape such as a quadrangle. In the integrated biochip of the present embodiment, a case where a plurality of reaction regions 140 are provided is described as an example. However, in some cases, only one reaction region 140 may be provided.

In the reaction region 140 of the present embodiment, a fluorescent antireflection film 150 is provided. Therefore, the surface of the anti-reflection film 150 may be the reaction region 140 itself or may be generated separately. The antireflection film 150 prevents the reflection of the fluorescence emitted from the sample 190. The anti-reflection film 150 may be disposed to cover the end of the excitation light absorption waveguide 120 exposed to the substrate 110. The anti-reflection film 150 may be formed of a material having affinity for the sample 190 or the liquid in which the sample 190 is dispersed so that the sample 190 may be attached well.

The anti-reflective film 150 is not an essential component of the present embodiment and may be omitted. When the anti-reflective film 150 is omitted, the surface of the reaction region 140 may be surface treated to have affinity for the sample 190 or the liquid in which the sample 190 is dispersed. This surface treatment may vary depending on the sample 190 to be detected. For example, when the substrate 110 is formed of a hydrophobic material such as silicon (Si), the reaction region 140 may be locally oxidized, an oxide may be applied, or a hydrophilic material may be applied to the hydrophilic material. Can have characteristics. Specific examples of such surface treatment do not limit the present embodiment. Depending on the sample 190, an ion exchange surface, an immobilized metal surface, or other various surface treatment methods may be applied.

The sample 190 attached to the integrated biochip of this embodiment is a sample that can be detected by a fluorescence detection method, and may be, for example, a biomaterial such as a fluorescently labeled nucleic acid. The biomaterial attached to the reaction zone 140 may be a detection probe or a target biomass combined with the detection biomaterial. Detection biomaterials are molecules that can interact with the target biomaterial, such as a hybridization reaction of a nucleic acid or an antigen-antibody reaction, for example, a nucleic acid molecule having a base sequence complementary to the nucleic acid molecule to be detected. have. On the other hand, the target biological material may be biological organisms such as enzymes, proteins, antibodies, nucleic acids, microorganisms, animal and plant cells and organs, nerve cells of the organism. For example, in the case of a DNA chip, as a detection biomaterial, a plurality of probe nucleic acids are attached to a predetermined reaction region 140 in a single spiral form. When a target nucleic acid (eg, mRNA) having a base sequence complementary to those of these probe nucleic acids is combined with a probe nucleic acid by binding, the fluorescent substance (L ') is emitted by the labeled fluorescent substance. Since the positions of the probe nucleic acids are predetermined, it is possible to determine the presence or absence of a plurality of target nucleic acids at the same time through the detected two-dimensional fluorescence image.

The plurality of excitation light absorption waveguides 120 are formed through the substrate 110. One end of the excitation light absorption waveguide 120 is placed on the side on which the sample of the substrate 110 is attached, and the other end of the excitation light absorption waveguide 120 is on the back side of the surface on which the sample of the substrate 110 is attached. . One end region of the excitation light absorbing waveguide 120 lying on the surface on which the sample of the substrate 110 is attached may correspond to the reaction region 140 to which the sample can be easily attached.

The plurality of excitation light absorption waveguides 120 includes a color filter core 121 and a cladding layer [1 (121] 123).

The color filter core 121 is formed of a material that transmits fluorescence emitted from a sample and absorbs excitation light that excites the sample. The color filter core 121 may have a circular or polygonal cross section. The shape of the cross section may correspond to the shape of the reaction region 140. In general, since the wavelength of the fluorescence is longer than the wavelength of the excitation light for exciting the fluorescence, the color filter core 121 may be a color filter material having a pass band of the fluorescence wavelength band. For example, the semiconductor, polymer or dielectric material of the transparent material may be dyed with a dye belonging to the wavelength range of fluorescence, or the pigment, which belongs to the wavelength range of fluorescence, may be formed on the transparent semiconductor, polymer or dielectric material. Since the dye or pigment used in such a color filter is well known in the display technology or the optical field, a detailed description thereof will be omitted.

The cladding layer 123 is formed of a material having a refractive index lower than that of the color filter core 121 so that the fluorescence passing through the color filter core 121 may totally reflect. For example, the cladding layer 123 may be made of a low refractive material such as MgF and various kinds of oxides such as SiO ₂ . In some cases, a dopant may be added to lower the refractive index of the substrate 110. Conversely, a dopant may be added to the color filter core 121 to relatively increase the refractive index of the color filter core 121.

The cladding layer 123 may have a multilayer structure having different refractive indices, or may have a structure in which the refractive indices gradually change. If the refractive index of the substrate 110 is smaller than that of the color filter core 121, the cladding layer 123 may be omitted, and the substrate 110 around the color filter core 121 may perform the function of the cladding layer. It may be. Meanwhile, since the refractive index of the color filter core 121 gradually changes, such as a graded index optical fiber, the refractive index may be continuously changed at the boundary between the color filter core 121 and the clad layer 123. It may be.

Referring to FIG. 2, the fluorescence L ′ emitted from the sample is generated by the excitation light L, and the downward fluorescence L ′ passes through the excitation light absorption waveguide 120 and the sample reaction part 100. ) It comes out to the back side. Meanwhile, the excitation light L is also directed downward but is absorbed and blocked while passing through the excitation light absorption waveguide 120. As described above, only the fluorescence L ′ is emitted when viewed from the rear side of the sample reaction unit 100, and thus, through the fluorescence image shown on the rear side of the sample reaction unit 100, it is possible to determine whether the sample 190 is detected. do.

The fluorescence L 'generated by the excitation light L is omnidirectional and therefore diverges in all directions. However, since the excitation light absorption waveguide 120 has a waveguide structure of an optical fiber, when the incident angle of incident light is out of a predetermined angle, the excitation light absorption waveguide 120 is lost in the excitation light absorption waveguide 120. Accordingly, the fluorescence L 'is guided only through the excitation light absorbing waveguide 120 under the reaction region 140 in which the sample 190 emitting itself is placed, so that the fluorescence L' of the neighboring sample 190 is guided. ) Can prevent the fluorescent image from being confused or blurred.

Referring back to FIG. 1, the sample detector 200 is provided on the rear side of the surface on which the plurality of reaction regions 140 of the substrate 110 are provided, and detects the fluorescence guided through the excitation light absorption waveguide 120. . The sample detector 200 includes a photodiode 210 provided with a plurality of photodiodes 220 and a wiring line 230 provided with a wiring line 239. The photodiode 220 corresponds to a pixel that detects light emitted from a sample. The sample detection unit 200 may be, for example, a substrate on which an image sensor such as a charge coupled device (CCD) or a CMOS image sensor (CIS) is provided. For example, in the case of the CIS, a CMOS circuit may be formed on the surface of the photodiode 210. The sample detector 200 may further include a signal processor (not shown) capable of processing the detected fluorescence.

The photodiode 220 is arranged so as to correspond one-to-one or one-to-many with the end of the excitation light absorption waveguide 120 in which fluorescence is emitted. One end of the excitation light absorption waveguide 120 meets the reaction region 140 of the substrate, and the other end of the excitation light absorption waveguide 120 is exposed to the rear surface of the substrate 110, so that the rear surface of the substrate 110 is the reaction region 140. ) Has a one-to-one correspondence with an array. Therefore, the image by the fluorescence emitted from the reaction region 140 side of the substrate 110 is displayed on the back surface of the substrate 110 as it is. Accordingly, the sample detection unit 200 may immediately read an image of the fluorescence emitted from the plurality of reaction regions 140 without additional optical members. Since the fluorescent image can be directly read from the integrated biochip itself, the integrated biochip of the present embodiment does not require a separate detection optical system, and the biodetection device can be miniaturized.

3 is a schematic perspective view of an integrated biochip according to another embodiment of the present invention. The integrated biochip of this embodiment has a structure in which a micro lens is further included in the integrated biochip of the above-described embodiment.

Referring to FIG. 3, the integrated biochip has a structure in which the sample reaction unit 101 and the sample detection unit 200 are integrally coupled.

The sample reaction unit 101 includes a plurality of micro lenses 160 provided at substrates 110, a plurality of excitation light absorption waveguides 120 penetrating through the substrate 110, and ends of the plurality of excitation light absorption waveguides 120, respectively. ). The excitation light absorption waveguide 120 includes a color filter core 121 and a cladding layer 123. The substrate 110 and the excitation light absorption waveguide 120 substantially correspond to the corresponding components of the above-described embodiment. Duplicate explanations will be omitted since they are the same.

The microlens 160 corresponds to the end of the excitation light absorption waveguide 120 one-to-one, and the surface of the microlens 160 becomes the reaction region 140 to which the sample is attached, and the microlens 160 of the region where the microlens 160 is provided. The outer edge is a non-reactive area to which no sample is attached. The surface of the microlens 160 or the outside of the region where the microlens 160 is provided is subjected to a surface treatment so that the sample can be attached to only the surface of the microlens 160. For example, when the substrate 110 is formed of a material that is not friendly to the sample or the liquid in which the sample is dispersed, the microlens 160 may be surface treated to have affinity for the sample or the liquid in which the sample is dispersed. This surface treatment may vary depending on the sample to be detected. For example, when the substrate 110 is formed of a hydrophobic material such as silicon (Si), the microlens 160 may be surface treated by oxidation or the like to have hydrophilic characteristics.

A fluorescence antireflection film (not shown) may be provided on the surface of the microlens 160 to suppress fluorescence from being lost on the surface of the microlens 160. As described above, the anti-reflection film may be formed of a material that is friendly to a sample or a liquid in which the sample is dispersed.

The microlens 160 has a convex shape and has a refractive power for condensing fluorescence emitted from the sample. Since the surface of the microlens 160 is convex, its width is wider than that of the flat case. Since the surface itself of the microlens 160 becomes a reaction region to which the sample is attached, when the microlens 160 having a convex shape is adopted as in the present embodiment, more samples can be attached to the same cross-sectional area, so that detection is possible. The intensity of fluorescence emitted from the sample to be increased may be increased.

In addition, the fluorescence incident on the excitation light absorption waveguide 120 may be transmitted when the total reflection condition is satisfied in the excitation light absorption waveguide 120. Thus, in general, the incident range of the light transmitted through the excitation light absorption waveguide 120 is limited. In the present exemplary embodiment, since the incident fluorescence is refracted by the microlens 160, the fluorescence incident more obliquely than the case without the microlens 160 satisfies the total reflection condition in the excitation light absorption waveguide 120. Can be. Accordingly, a larger amount of fluorescence emitted from the sample may be transmitted to the excitation light absorption waveguide 120.

In the present embodiment, the microlens 160 has a convex hemispherical shape as an example, but is not limited thereto. Since a sample, such as a target biomaterial, flows through the surface of the integrated biochip dispersed in a liquid, the refractive index of the liquid in which the sample is dispersed may be greater than that of the microlens 160. As such, when the refractive index of the liquid in which the sample is dispersed is relatively higher than the refractive index of the microlens 160, the microlens 160 may have a hemispherical shape concave on the surface of the substrate 31.

On the other hand, the fluorescence proceeds in the excitation light absorbing waveguide 120 and leaves the excitation light absorbing waveguide 120, so that the cross-sectional area of the light beam is gradually increased. Rayleigh length means the distance from the point where the light beam is focused and its cross-sectional area is minimum to the point where its cross-sectional area is doubled. Therefore, the photodiode 220 of the sample detecting unit 200 may be arranged at approximately Rayleigh length from the end of the excitation light absorption waveguide 120 to design the size of the sample detecting unit 200 to increase the fluorescence detection efficiency. Can be.

4 illustrates an integrated biochip in accordance with another embodiment of the present invention. Referring to FIG. 4, the biochip 300 of the present embodiment is packaged, and the integrated chip 310 is mounted on the frame 330. The integrated chip 310 means that the sample reaction unit (100 of FIG. 1) and the sample detection unit (200 of FIG. 1) are bonded to each other in the above-described embodiment. The upper surface of the integrated chip 310 is a surface provided with a reaction region (140 of FIG. 1) to which a sample may be attached, and may be exposed to the outside. For example, in the case of a DNA chip, a plurality of probe nucleic acids are fixed to the reaction region 140 of the integrated chip 310 through a semiconductor process or the like, and when a liquid containing the target nucleic acid is flowed onto the surface of the DNA chip, Probe nucleic acids having a base sequence complementary to the nucleic acid sequence of the nucleic acid is combined by the reaction mixture with the target nucleic acid, nucleic acids that are not combined with the probe nucleic acids of the DNA chip is washed out. Since the nucleic acid subjected to the mixing reaction emits fluorescence by the labeled fluorescent substance, the presence or absence of the target nucleic acid to be detected is determined by detecting the position where the fluorescence is emitted. As described above, the biochip of the present embodiment can detect fluorescence emitted by the chip itself, and thus it is possible to determine whether the sample is detected through an electrical signal output from the packaged biochip 300.

5 illustrates an integrated biochip in accordance with another embodiment of the present invention. Referring to FIG. 5, the biochip 301 of the present embodiment is packaged, and includes an integrated chip 310, a frame 330 on which the integrated chip 310 is mounted, and a cover glass for protecting the integrated chip 310. 340). This embodiment is substantially the same as the above-described embodiment except that the cover glass 340 is further provided.

The cover glass 340 may be detachably installed, or a passage (not shown) may be provided on one side of the cover glass 340 to allow the fluid containing the sample to flow in and out. The surface of the cover glass 340 may be coated with an excitation light reflection prevention film 341 to prevent reflection of the excitation light for sample detection. The cover glass 340 prevents surface damage of the integrated chip 310, which may be caused in actual use of the biochip 301.

Next, FIGS. 6A to 6E, 7, and 8A to 8C will be described a manufacturing process of an integrated biochip according to an embodiment of the present invention.

First, a substrate 110 as shown in FIG. 6A is prepared. The substrate 110 may be formed of glass, a semiconductor, a metal, a dielectric material, or a polymer. As the material of the substrate 110, a material that is not friendly to the sample or the liquid in which the sample is dispersed may be selected. For example, the substrate 110 may be a silicon substrate having hydrophobicity. One surface of the substrate 110 may be planarized by forming a flat layer or by a process such as chemical mechanical polishing (CMP).

Next, as illustrated in FIG. 6B, a plurality of through holes 110c are formed in the substrate 110. The diameter of the through hole 110c may have a size of sub μm to several tens of μm. TSV process technology for forming through holes having a diameter of several μm in a silicon substrate having a thickness of several tens of micrometers or more is known in the art. Such micro-sized through holes 110c are, for example, TSVs (Through Silicon Via). ) Can be formed through the process.

If the diameter of the through hole 110c is made very small, the depth of the through hole 110c may not be sufficient, and the through hole 110c may not penetrate the substrate 110. In this case, the through-hole 110c is formed through a back-lap process of polishing the back surface 110b of the surface 110a on which the incomplete through-hole 110c is formed to the depth of the through-hole 110c. 110 may be penetrated. Such a backlap process may be performed, for example, after forming an excitation light absorption waveguide described later or after surface treatment of the substrate.

Referring to FIG. 6C, the cladding layer 123 is formed by oxidizing the inner wall surface of the through hole 110c of the substrate 110 to reduce the refractive index. The refractive index can be controlled by controlling the degree of oxidation or by further diffusing the dopant. In the cladding layer 123, a dopant may be injected into the through hole 110c to diffuse the dopant into the inner wall surface of the through hole 110c, or a low refractive index material may be formed on the inner wall surface of the through hole 110c. The method of coating may be adopted.

Referring to FIG. 6D, the color filter core 121 is formed by filling a color filter material into the through hole 110c in which the clad layer 123 is provided. The color filter material is a dye or pigment in the fluorescence wavelength band is combined with a transparent binder, and a material having a refractive index larger than that of the cladding layer 123 is selected. After filling the color filter material, the surface of the substrate 110 may be planarized through a process such as CMP.

Referring to FIG. 6E, a fluorescent antireflection film 150 is formed on a portion of the clad layer 123 and the color filter core 121 exposed to the surface of the substrate 110. The anti-reflection film 150 may be formed using a photolithography process.

Prior to forming the anti-reflective film 150, a process of forming a microlens (see 160 of FIG. 3) on the exposed portion of the clad layer 123 and the surface of the substrate 110 of the color filter core 121 may be added. Can be. The microlenses may be formed by, for example, forming a pattern of a microlens array with photoresist and then deforming the columnar patterned photoresist into a curved photoresist through a reflow process. In addition, the surface of the cladding layer 123 and the surface of the substrate 110 of the color filter core 121 in place of the fluorescent antireflection film 150 is surface treated to increase affinity for the sample or the liquid containing the sample. It may be.

Meanwhile, referring to FIG. 7, the sample detector 200 including the photodiode 220 is provided. The sample detector 200 may be a CCD or a CIS. For example, in the case of a CIS, a photodiode 220 is formed on a silicon substrate 210 by a frontend process, and a wiring line part 230 having wiring lines 239 including a CMOS circuit is formed thereon. do. Next, a CIS substrate is manufactured by forming vertical and horizontal passive metal layers connecting circuits formed on the silicon substrate 210 by a backend process. In a typical manufacturing process of an image sensor, a process of forming a color filter on the CIS substrate is further added, but the sample detecting unit 200 of the present embodiment does not further include a process of forming such a color filter. Since the sample detecting unit 200 of the present embodiment is substantially the same as a conventional image sensor and can be manufactured through the same process, detailed description thereof will be omitted. For example, in the case of a CCD, a photodiode 220 that generates electrons and a horizontal vertical metal layer that enables electron transmission are connected through a wiring line unit 230. The photoelectron is transported to the periphery of the sensor through the field transfer device of the metal layer and then converted into a voltage.

On the surface 230a of the completed sample detection unit 200, a planarized layer is formed or the surface 230a is evenly made by using CMP. In addition, an alignment mark may be formed on the surface 230a for the wafer bonding process described later.

Next, as illustrated in FIG. 8A, the sample reaction unit 100 and the sample detection unit 200 are bonded to each other. Such bonding can be done on a wafer basis or on an individual chip basis. For example, the sample reaction unit 100 substrate and the sample detection unit 200 substrate may be aligned using an alignment mark, and the like, and may be directly bonded in a wafer unit. Such direct bonding may be achieved by applying a predetermined pressure to the aligned substrate to fuse the bonding surface or by using plasma.

Next, referring to FIG. 8B, an area where the excitation light absorption waveguide 120 is formed may be surface treated so that the sample may be attached to the surface of the sample reaction part 100 bonded to the sample detection part 200. Such surface treatment may be performed before the sample reaction unit 100 and the sample detection unit 200 are bonded. In addition, if the anti-reflection film 150 itself has a property that can be attached to the sample well, the surface treatment step may be replaced by forming the above-described anti-reflection film 150.

Next, referring to FIG. 8C, a process of attaching the sample 190 to the sample reaction unit 100 may be performed. The sample 190 may be a probe for detecting a biomaterial that can interact with a target biomaterial to be detected. For example, DNA bases such as A (adenine), G (guanine), C (cytosine), and T (thymine) may be stacked in a different order for each reaction region 140 by using a photo-lithography process. A DNA chip to which a probe nucleic acid having a sequence is attached can be prepared.

Next, each chip is divided and a wire bonding process is performed to complete the biochip package.

Hereinafter, a detection apparatus using an integrated biochip according to the above embodiments will be described.

9 illustrates a schematic configuration of a bio detection device according to an embodiment of the present invention.

Referring to FIG. 8, the biodetection apparatus of this embodiment includes a light source 501, a light diffusing element 502, a collimating lens 504, and a condensing lens, which are illumination optical systems that irradiate the integrated biochip 300 with excitation light. 507. Reference numeral 520 denotes a stage in which the integrated biochip 300 is detachably installed.

The light source 501 emits excitation light (L). The excitation light L is light for exciting the fluorescent material attached to the biomaterial in the integrated biochip 300. Typically, light having a wavelength of approximately 500 nm is used as the excitation light, but depending on the fluorescent material to be labeled, the wavelength of the excitation light L may vary. The wavelength of the excitation light L emitted from the light source 501 may be shorter than the fluorescence. If a white light source is used as the light source 601, an excitation filter (not shown) that blocks light of a wavelength band such as fluorescence on the optical path between the light source 601 and the integrated biochip 300 is used. To place.

The light diffusing element 502 evenly diffuses the excitation light L to have a uniform intensity through its entire cross section, and may be, for example, a bar-shaped light integrator. The excitation light L has an overall uniform intensity to illuminate light having the same intensity over a portion or the entire region of the integrated biochip 300.

The collimating lens 504 shapes the excitation light L in parallel. Although FIG. 8 is shown to be disposed between the optical diffusion element 502 and the condenser lens 507, the collimating lens 504 may be disposed between the light source 501 and the light diffusion element 502. Further, when the divergence of the excitation light L emitted from the light source 501 is not large and the excitation light L can be sufficiently focused by the condenser lens 507, the collimating lens 504 is used. You may not.

The condenser lens 507 condenses the excitation light L to provide a light spot having a predetermined diameter on the integrated biochip 300. The diameter of the light spot may be large enough to illuminate a part or the whole area of the integrated biochip 300.

When the spot of the excitation light L covers the entire surface of the integrated biochip 300, the integrated biochip 300 may simultaneously detect the fluorescent image emitted from the sample. If the spot of the excitation light L covers only a part of the integrated biochip 300, the stage 520 or the illumination optical system moves to cover the entire surface of the biochip 300 in which the excitation light L, which is illuminated, moves. In order to cover sequentially, the integrated biochip 300 obtains fluorescence images of the entire reaction region to which the sample is attached by synthesizing the images of fluorescence emitted sequentially.

The integrated biochip 300 of the present embodiment directly detects the fluorescence emitted from the sample by the illuminated excitation light L, and the signal for the detected fluorescence is transmitted to the signal processing system not shown through the stage 520. do. The biodetector of the present embodiment can read an image of fluorescence emitted from a sample as the integrated biochip 300 itself without having to provide a separate detection optical system. Therefore, since the optical system of the bio-detection device is sufficient only as the illumination optical system, it is easy to manufacture portable, and it is possible to analyze and detect samples such as biomaterials regardless of the place.

The present inventors integrated biochip and its manufacturing method have been described with reference to the embodiments shown in the drawings for clarity, but this is merely illustrative, and those skilled in the art will appreciate that various modifications and equivalents therefrom It will be appreciated that other embodiments are possible. Therefore, the true technical protection scope of the present invention will be defined by the appended claims.

1 is a schematic cross-sectional view of an integrated biochip according to an embodiment of the present invention.

FIG. 2 is an enlarged cross-sectional view of one excitation light absorption waveguide of the sample reaction part of FIG. 1.

3 is a schematic cross-sectional view of an integrated biochip according to another embodiment of the present invention.

4 is a schematic perspective view of an integrated biochip according to another embodiment of the present invention.

5 is a schematic cross-sectional view of an integrated biochip according to another embodiment of the present invention.

6A to 6E, 7, and 8A to 8C illustrate a method of manufacturing an integrated biochip according to an embodiment of the present invention.

9 is a schematic configuration diagram of a bio detection device according to an embodiment of the present invention.

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

100, 101 ... sample reaction part 110 ... substrate

120 ... excited light absorption waveguide 140 ... reaction zone

150 ... Fluorescent reflector 160 ... Microlens

190.Drugs 200 Sample detection unit

210.Photodiode section 220.Photodiode

230 ... wiring line part 300, 301 ... package

310.Biochip 330.Frame

340 Cover glass 341

501 ... light source 502 ... light diffusing element

504 ... collimating lens ...

520 stage

Claims (18)

A substrate provided with at least one reaction region to which a sample is attached, and absorbing excitation light provided in each of the at least one reaction region to excite the sample and transmitting fluorescence emitted from the sample. A sample reaction part having at least one excitation light absorption waveguide penetrating from the prepared substrate surface to the rear surface of the substrate surface; And And a sample detection unit provided on a rear side of the substrate surface on which the plurality of reaction regions of the substrate are provided to detect fluorescence emitted from the sample, the sample detection unit integrally coupled with the sample reaction unit. According to claim 1, The excitation light absorbing waveguide includes a color filter core that transmits fluorescence emitted from a sample and absorbs excitation light that excites the sample. The method of claim 2, And the color filter core has a refractive index higher than that of the substrate surrounding the color filter core. The method of claim 2, The excitation light absorption waveguide is provided around the color filter core and further comprises a cladding layer having a refractive index lower than the refractive index of the color filter core. The method according to any one of claims 1 to 4, And a micro lens configured to condense fluorescence emitted from a sample in the plurality of reaction regions. The method according to any one of claims 1 to 4, The biochip integrated in the plurality of reaction zones, the anti-reflective film for transmitting the fluorescence emitted from the sample. The method according to any one of claims 1 to 4, The sample detection unit is an integrated biochip that is a charge coupled device (CCD) or a CMOS image sensor (CIS) The method of claim 7, wherein And the pixel of the sample detecting unit corresponds one to one or one to many with the at least one reaction region. The method according to any one of claims 1 to 4, Integrated biochip further comprises a frame for mounting the sample reaction unit and the sample detection unit. The method of claim 9, Protecting the sample reaction unit and the sample detection unit, the integrated biochip further comprises a cover glass antireflective coating for the excitation light. Providing a sample reaction part having at least one excitation light absorption waveguide formed through the substrate; Providing a sample detector including at least one pixel; And And bonding the sample reaction unit and the sample detection unit to the integrated biochip. 12. The method of claim 11, Preparing the sample reaction unit, Forming at least one through hole in the substrate; And Filling a color filter material into each of the plurality of through holes to form a color filter core. The method of claim 12, Preparing the sample reaction unit, And forming a cladding layer on an inner wall of the plurality of through holes before forming the color filter core. The method of claim 12, Preparing the sample reaction unit, And forming a microlens in an area of the substrate on which an excitation light absorption waveguide of the substrate is formed. 12. The method of claim 11, The sample detection unit is a manufacturing method of an integrated biochip is a CCD (Charge Coupled Device), or a CMOS Image Sensor (CIS) substrate. The method according to any one of claims 11 to 15, Bonding the sample reaction unit and the sample detection unit, And a pixel of the sample detecting unit corresponding to the at least one excitation light absorption waveguide in a one-to-one or one-to-many manner. The method according to any one of claims 11 to 15, Bonding of the sample reaction unit and the sample detection unit is a manufacturing method of an integrated biochip made in a wafer unit or an individual chip unit. The method according to any one of claims 11 to 15, And surface treating a region where the excitation light absorption waveguide is formed so that a sample may be attached to one surface of the sample reaction unit.

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