TW200408711A - Cell chip for bio detection and method for producing the same - Google Patents

Abstract

A cell chip for bio detection includes a housing region installed corresponding to the size of each test specimen having an entry opening for a test specimen solution installed on top of the housing region, and an extraction opening for extraction of non-specimen; a passivation layer covering the surface of the housing region for patching and passivation of the housing region and providing a background for optical detection; and a detection layer formed by covering the surface of the passivation layer with a bio-compatible polymer so that each test specimen is accurately positioned in a single housing region for a subsequent detection.

Description

200408711 V. Description of the invention (1) The present invention relates to a bio-assay cell wafer, especially a bio-chip used to detect the specific specimen location of a cell. Biochips are designed using the principles of molecular biology, genetic information, analytical chemistry, etc., using silicon wafers, glass or polymer organic plastics as carrier plates, and cooperating with micro-electromechanical or other precision processing technologies. Responsive, accurate and cost-effective bioanalytical testing capabilities. The chip function is often used in advanced countries in Europe and the United States to accelerate life science research and shorten the development time of new drugs. The concept of biochip originated in the 1980s. According to its characteristics, biochip can be divided into gene chip, process chip (lab-on chip) and protein chip: 1. Gene chip (Gene chip): A chip made by placing thousands to tens of thousands of different gene segments (called nucleic acid probes) on a carrier board with an area of several square centimeters. Place the wafer in the test liquid during use to hybridize the nucleic acid probe on the wafer with the test gene in the liquid (Hybridizati ο η). If a certain gene has a hybridization reaction, it will show a different fluorescent color It can be distinguished by naked eyes or special instruments. Its advantage is that it can provide a large variety of different gene sequence related information through one test, eliminating the time-consuming and labor-intensive situation that can only detect one gene performance at a time, greatly saving time and procedures, and greatly helping to accelerate the interpretation of gene information. beneficial. The most important and basic principle of a gene chip is to use the characteristics of the hybridization reaction of a gene to achieve the purpose of identifying the gene. 2. Protein chip: Proteins are used as biological probes, which are arranged neatly on the chip to carry out the antigen-antibody immune reaction for

200408711 V. Description of the invention (2) Detection of protein. 3. Process chip (lab-on chip): The purpose is to miniaturize a certain experimental environment and equipment in a biochemical laboratory, which is equivalent to transposing the biochemical experiment on the wafer, so a photomask is used in the microfluidic wafer. Etching many capillary channels, the inner diameter of the pipeline is about several hundred micrometers or even several micrometers, and the micro-electromechanical technology is used to pump the liquid to be tested into the capillary channels, so that the liquid is separated and mixed in the designed environment. Mixed), culture (Incubation), heating (Heated), polymerase chain reaction pCR (Polymerase Chain Reaction) and other laboratory reactions. In order to analyze a large number of cells or genes in parallel and in parallel, many researchers have racked their brains to study how to effectively locate the object on a biochip. The most commonly used method is microarray technology. The robotic arm is used to fix the bioprobe to the biochip in a high-density array mode, and then the test object is contacted with the bioprobe by the extraction needle for detection and analysis. However, in this way, a bioprobe is required for each array point. And the extraction needle, the components are too complicated, and using the extraction needle to suck the object to be tested can easily cause damage to the object and the biological probe. The other way is to set the reaction area of the fixed biological probe and the micro-channel for sample solution injection and extraction. Although this configuration can effectively isolate the sample from contact with the outside world, it can achieve good experimental control without damage. Specimens, but due to cost and technical considerations, only a few reaction zones can be arranged on a biochip, which is difficult to detect in large numbers, and multiple specimens may be adsorbed by probes in a single reaction zone for subsequent fluorescent detection. Due to each

200408711 V. Description of the invention (3) The activity and biological characteristics of the specimens are different, which will affect the detection accuracy. That is, the main purpose of the present invention is to solve the above-mentioned shortcomings and avoid the existence of the shortcomings. The present invention is based on the bio-detection wafer with an accommodating area according to a specific specimen size, and a passivation layer and The detection layer enables each specimen to be accurately positioned in a single accommodating area for subsequent detection. The detailed description and technical contents of the present invention are described below with reference to the drawings:

Please refer to "shown in Figures 1, 2-1 and 2-2", which is the manufacturing process, three-dimensional appearance and partial enlarged cross-sectional schematic diagram of the present invention, as shown in the figure: The aforementioned method for manufacturing a biochip includes the following steps ： Define the specimen accommodating space 1, which is on a silicon wafer with a length and width of 2 cm each. The wafer is etched and arrayed (100 points each for length and width, 10,000 points in total). The microbe detection wafer 10 defines a downwardly tapered specimen holding space. Above the containing space is a drop inlet 1 1. The aforementioned etching technology may be wet etching or dry etching, and wet etching is performed by chemical reaction. Dry etching is performed by physical action;

After the specimen retention space is completed, the non-specimen extraction opening 2 is cut, and the bottom of the retention space is cut according to the specific specimen size, so that the extraction space 12 is formed at the bottom of the retention space, which is defined by the aforementioned drip inlet 11 and extraction opening 12. A accommodating region 13 with a wall facing downward is covered; the passivation layer 3 is covered, and after the accommodating region 13 is completed, a passivation layer 14 is formed on the surface of the accommodating region 13 in order to further precisely control the size of the accommodating region 13. The passivation layer 14 is a metal chromium plated on the surface of the accommodating region 13.

Page 8 200408711, description of the invention (4) ^ 〇 / N (a) through the metal purification layer 14 can repair the aforementioned accommodating area (two, the contact angle of the brother, and further precisely control the aperture of the extraction port 12 , ^, In the fluorescence detection of the detection object, it is best not to generate light in the background and affect the scanner CCD (charge-coupled device) or co-…, radiance (c ο nfoca 1 1 asersca η ning) The detection value is ideally a black array (b 1 ackmatrix), which reduces the optical detection error through the low reflectivity of chromium; after the passivation layer is completed, in order to avoid the impact of the passivation layer 14 on the specimen, resulting in detection errors, slope Covering the detection layer 4, the detection layer 15 deposits a biocompatible polymer on the surface of the passivation layer 14; the biocompatible polymer is a parylene (pylene-p-ly-p- xyiyiene) is formed by deposition after the process of vaporization (150 oC) and cracking (650 ° C). The advantage of polyparaben is that it has good coverability and can completely isolate the underlying passivation layer 1 4. Please Refer to "as shown in Figures 3 and 4". The schematic diagram of the embodiment of the invention is as shown in the figure: the bio-detection wafer 10 can be assembled on a base 20, and the base 20 includes a positioning area 21 for the bio-detection wafer 10 to be accommodated and located at the location. The area 2 1 below and on both sides are respectively connected to a pumping channel 22 (not shown) of the pump. When in use, the biometric chip 10 is placed in the positioning area 21 of the base 20, and the base 20 The extraction channels 22 are respectively connected to the pump. At this time, the detection system configuration is completed. When the experiment is performed, the sample solution is dropped from the drip inlet 11 into the aforementioned biological detection wafer 10, and then the pump is started to remove impurities in the sample solution. And the water is extracted through the lower extraction port 12 and the extraction channel 22, and at the same time, because the pump provides a pressure difference, the specimen can be more firmly located in the biological examination

Page 9 200408711 V. Description of the invention (5) In the accommodation area 1 3 (see Figure 2) of the test chip 10, turn off the pump, and then each specimen is positioned in each accommodation area 13 respectively. , You can perform subsequent tests. Since the accommodating area 13 of the present invention is used to locate a specimen of a specific size, the aperture of the extraction port 12 must be set according to the size of the specimen. For details of the dimensions of common specimens, see Annex 1, the most commonly used for detection Samples of human diseases include ... liver cells, red blood cells, and bacteria, the size of which is between 100 nm-1 0 // m (micron size), so if you want to detect such cells, the sample of this creation The pore size of the exit port 12 must be less than 100 nm. If you want to detect a larger (plant cell) or smaller (nano size such as virus, protein or DNA) sample, you only need to change the pore size of the exit port 12. The specific specimen can be positioned in the accommodating area 1 3 according to the size. As for impurities or water with a smaller size, the sample is separated from the accommodating area 13 under the extraction port 12. However, the above are only the preferred embodiments of the present invention. When the scope of implementation of the present invention cannot be limited, that is, all equal changes and modifications made in accordance with the scope of the patent application of the present invention shall still belong to the patent of the present invention. Covered.

Page 10 200408711 Brief Description of Drawings [Simplified Description of Drawings] Figure 1 is a schematic diagram of the manufacturing process of the present invention. Figure 2-1 is a perspective view of the present invention. Figures 2-2 and 2-1. Partial enlarged cross-sectional schematic diagram 3, which is a schematic diagram of an embodiment of the present invention, FIG. 4, which is a schematic cross-sectional diagram of an embodiment of the present invention, is a size distribution chart of common specimens [Description of symbols in the diagram] Definition inspection Body accommodating space ............ Cutting non-specimen extraction opening ......... Covering purification layer ............... ... 3 Overlying detection layer ........ 4 Biological detection chip..10 ..... 11 Evacuation port ..... 12 Receiving area ... ........ 13 Pure 4 Dagger Layer ......... 14 Detection Layer ... 15 Base ... 20 Positioning area ... ............. 21 Evacuation Channel ... 22

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Claims (1)

200408711 VI. Application Patent Scope 1. A bioassay cell wafer, the bioassay cell wafer includes: an accommodating area, the accommodating space is set corresponding to the size of each specimen, and a specimen fluid is arranged above the accommodating area. The drip inlet is provided with an extraction opening that is not subject to sample isolation; a passivation layer covering the surface of the aforementioned containing area to repair and passivate the aforementioned containing area and provide a background for optical detection; and a detection layer Is coated on the surface of the passivation layer with a biocompatible polymer. 2. The bioassay cell wafer as described in item 1 of the scope of the patent application, wherein the bioassay cell wafer may be a wafer 2 cm in length and 2 ° in length. 3. The bioassay cell as described in the scope of patent application item 2 The wafer, wherein the accommodating area is arranged in an array manner with a length and a width of more than 5 points each on the bioassay cell wafer. 4. According to the bioassay cell wafer described in item 1 of the scope of the patent application, the drop inlet aperture of the containing area is larger than the extraction opening, so that the wall surface of the containing area is tapered downward. 5. The bioassay cell wafer according to item 1 of the scope of the patent application, wherein the passivation layer is made of metal chromium or chromium oxide, which can repair the sharp contact angle of the passivation containing area and oxidize by chromium or chromium The low reflectivity of objects to light provides a background for optical detection. 6. The bioassay cell wafer according to item 1 of the patent application scope, wherein the biocompatible polymer layer is, for example, polyparaphenylene 200408711 6. Scope of patent application (Parylene, poly-p-xylylene), etc., so that the contact between the wafer and the specimen will not affect the activity of the specimen. 7. The bioassay cell wafer according to item 1 of the scope of the patent application, wherein the bioassay cell wafer can be assembled on a pedestal, and the base includes a positioning area for accommodating the bioassay cell wafer and is located at the location. Below and on both sides of the zone are connected to the pumping extraction channel. 8. The bioassay cell wafer according to item 1 of the scope of the patent application, wherein the aperture of the extraction port directly corresponds to the size of the specimen, and the aperture of the extraction port is slightly smaller than the specimen. 9. The bioassay cell wafer according to item 1 of the scope of the patent application, wherein the size of the specimen can be divided into centimeters, micrometers and submicrons: plant cells, animal cells, general blood cells and bacteria. 1 0. — A method for manufacturing a biological detection cell wafer, including the following steps: a) Define a specimen accommodating space, and form a downwardly tapered inspection on the biological detection wafer by wafer etching according to the size of the specimen The body accommodation space, a drop inlet above the accommodation space; b) cutting the non-subject extraction opening, cutting the bottom of the retention space according to the specific specimen size, so that the bottom of the retention space forms a space for non-subject separation The extraction opening defines a receiving area with a wall facing downward by the aforementioned drip inlet and the extraction opening; c) a passivation layer is coated to form a passivation on the surface of the aforementioned receiving area in order to precisely control the size of the receiving area; Layer; and d) covering the detection layer to prevent the activity of the specimen from being affected. Page 13 200408711 VI. Scope of patent application A biocompatible polymer layer is formed on the surface of the chemical layer. 11. The method for manufacturing a biological detection cell wafer as described in item 10 of the scope of the patent application, wherein the biological detection cell wafer may be a wafer having a length and a width of 0.2 cm or more each. 12. The method for manufacturing a bioassay cell wafer as described in item 10 of the scope of the patent application, wherein the reserved space is arranged on the bioassay cell wafer in an array of more than 5 points in length and width each. 13. The method for manufacturing a biological detection cell wafer as described in item 10 of the scope of the patent application, wherein the wafer etching may be wet etching or dry etching. 14. The method for manufacturing a bioassay cell wafer as described in item 10 of the scope of the patent application, wherein the passivation layer can be electroplated with metal chromium on the surface of the accommodation area to repair the sharp contact angle of the passivation accommodation area, and by The low reflectivity of chromium or chromium oxide to light provides a background for optical inspection. 15. The method for manufacturing a bioassay cell wafer as described in item 10 of the scope of the patent application, wherein the detection layer is poly (p-xylene) (parylene, po 1 yp-xy 1 y 1 ene) and is deposited in a deposition manner. > formed on the surface of the passivation layer, so that the wafer's contact with the specimen will not affect the activity of the specimen Page 14