KR20040041823A - Programmable mask for forming biomolecule or polymer array and Fabrication method of biomolecule or polymer array using the same - Google Patents

Abstract

PURPOSE: A programmable mask for forming a biomolecule or polymer array and a method for producing the biomolecule or polymer array using the same are provided, thereby easily and cheaply forming the array with high density, and producing the biomolecule or polymer with high purity. CONSTITUTION: A programmable mask for forming a biomolecule or polymer array comprises an unit pixel of array and a driver circuit portion for selectively supplying the electric power to each pixel, wherein the each unit pixel comprises a solution containing charged particles inhibiting the progression of incident light and moved by electrophoresis, and an electrode for supplying the electric power to the particles for controlling the permeability of the incident light by change of the particle arrangement, wherein the solution is suspending fluid consisting of fluid and numerous charged particles; and the particles are barium sulphate, kaolin, zinc oxide or TiO2 particles with a size of 500 to 3000 angstrom.

Description

Programmable mask for forming biomolecule or polymer array and method for forming biomolecule or polymer array using same

The present invention relates to a programmable mask for forming a biomolecule or polymer array, and more particularly, to a transmissive programmable mask for forming a biomolecule or polymer array such as DNA by irradiating UV light to a specific cell and a biomolecule or polymer array using the same. It relates to a forming method.

First, a region of one type of biomolecule or polymer in a biomolecule array formed on a substrate will be referred to as a cell.

A lot of researches are being carried out to perform various kinds of experiments at once using biomolecules or polymer arrays. Examples of biomolecule or polymer arrays include arrays of polypeptides, carbohydrates and nucleic acids (DNA, RNA). The most important thing for this study is to form a high-density array of high purity at low cost on the substrate.

Until now, the method of forming a biomolecule or polymer array has been spotted in which a micro robot moves three-dimensionally and selectively drops a biochemical at a desired position, and adjusts the electrode voltage of the microelectrode array. Electronic addressing method that allows biomolecules to be fixed to specific electrodes, and photolithography that changes the surface by selectively irradiating light at a desired location so that the reaction between the surface and the biomolecules occurs only at a specific location And the like.

Spotting methods include a contact printing method in which a solution is buried like a paper, and a non-contact printing method in which the solution is dropped. The contact printing method is performed in the order of loading, printing and washing by the XYZ robot. Like a fountain pen nib, a grooved pin can be used to adjust the volume of the sample reproducibly and print multiple times with one sample load. However, there is a disadvantage in that the number of arrays per unit area cannot be increased.

Non-contact printing methods include dispensing and ink-jet printing. Dispensing is the dropping of a solution, as in the case of using a micropipette, and ink jet printing is a method in which the solution is ejected by applying a slight pressure to the reservoir. Ink jet printing allows for very fine control of the sample solution down to the nanoliter level, which greatly increases the number of arrays per unit area. Although an ink bottle is required for each sample solution, since the number of ink bottles that can be mounted on the robot is limited, there is a problem that it can be used only when an array is formed using only a few sample solutions.

Electronic addressing method is a method of fixing biomolecules using the voltage control function of the microelectrode array, which causes the charged biomolecules to move to the electrode surface and cause physicochemical bonding to occur. (Cosnier, Serge, "Biomolecule immobilization on electrode surfaces by entrapment or attachment to electrochemically polymerized films. A review" Biosensors & Bioelectronics 14 : pp. 443-456 (1999). For example, because DNA has a strong negative charge, when the electrode has a positive charge, the DNA moves toward the electrode. At this time, if a physicochemical bond occurs between the DNA and the electrode, the DNA is fixed to the electrode (US Patent No. 5,605,662). Such an electronic addressing method is difficult to apply when the number of arrays is large, and there is a problem that a microelectrode array is basically required. In addition, a method of electrochemically positioning biomolecules by electrochemically changing pH around the electrode has been developed. Combimatrix uses this concept to propose a method for synthesizing oligonucleotides on microelectrodes selectively (US patent). Publication No. 6,090,302). This method has a problem in that the yield of each reaction is low and the purity of each cell is very low.

On the other hand, the optical lithography method used in the semiconductor process was first used by Affymetrix (US Patent No. 5,959,098). This method has the advantage of being able to make dense arrays and parallel synthesis. However, since a large number of photomasks are required, there is a problem in that a large cost and time are required.

A method of using a programmable mask that can control the light path for each pixel without using a photomask has also been developed (US Patent No. 6,271,957). The programmable mask has a method of controlling light reflection or a method of controlling light transmission. An example of controlling light reflection is using a micromirror array. An example of controlling light transmission is LCD ( liquid crystal displays). The method of forming the array through the mirror manipulation of the micromirror array requires a complicated optical system and has the disadvantage that only a mosaic pattern can be obtained (US Patent No. 6,271,957 and US Patent No. 6,375,903).

Transmissive programmable masks have a relatively simple optical system compared to reflective ones, but have difficulty in increasing the contrast ratio corresponding to the ratio of light passing through and out of transmission. If the contrast ratio is low, UV light passes through the unwanted pixels and the photoreaction occurs in the cell. Therefore, a high purity biomolecule or polymer array cannot be obtained. In the LCD programmable mask, the corresponding substrate is controlled by controlling the light transmission of each pixel. Repeating the photoreaction to the cells in the phase to form a biomolecule or polymer array (US Patent No. 6,271,957, Korean Patent Publication No. 2001-0002915). However, the LCD programmable mask has a problem in that UV transmittance is low and is denatured by UV. While typical LCDs transmit light in the visible range, UV light with a wavelength of 400-330nm is required to drop the protector. In the case of the liquid crystal and the alignment film in the general LCD, UV light tends to be denatured.

On the other hand, the method of controlling the transmission of light has been much research in the display (display) field. In particular, an electronic ink technology has been developed that transfers charged particles to electrophoresis to cause color change (US Pat. No. 6,120,588). This technique cannot be used for transmissive purposes because it changes the color of the reflected light. This technique is also unsuitable for use in obtaining biomolecules or polymer arrays because of the small contrast ratio of permeation to opacity.

In order to obtain a high-density biomolecule or polymer array, a high-density programmable mask is required. As in the general display field, a high-density programmable mask can be obtained by converting each pixel into light transmission or opacity by a transistor.

Accordingly, an object of the present invention is to provide a new programmable mask capable of easily and cheaply forming a high purity, high density biomolecule or polymer array.

One aspect of the present invention for achieving the above object is a programmable mask including an array of unit pixels and a driver circuit for selectively applying a voltage to each of the unit pixels, to adjust the transmittance of incident light for each unit pixel Each of the unit pixels has a solution including charged particles that obstruct the progress of incident light and move by electrophoresis, and a voltage is applied to the particles to change the arrangement of the particles to control the transmittance of the incident light. There is provided a programmable mask having electrodes for it.

The "programmable mask" means any device that is configured to control the transparency / opacity of incident light on a unit pixel basis. In addition, if necessary, not only transparent / opaque but also a gray level structure can be implemented. On the other hand, the switching element for selectively applying a voltage to the unit pixels may be any device that can perform a switching role without particularly limited in scope, such as a thin film transistor, a MIM element. The driver circuit unit may be implemented on a substrate, such as a pixel, or may be separately implemented on a printed circuit board.

The solution may also be a suspension fluid consisting of fluid and a number of charged particles, the fluid being fluorocarbon, chlorocarbon, fluorocarbon ( fluorochlorocarbon) or trichlorofluoroethylene, and charged particles are TiO ₂ particles with a size of 500-3000 mm ₃ . On the other hand, the charged particles may block or pass the UV light by absorbing or reflecting the UV light.

Preferably, the charged particles accumulate on the electrode selected by the voltage difference of the electrodes, and when the charged particles accumulate on the electrode perpendicular to the incident direction of the incident light, the incident light is blocked and is parallel to the transmission direction of the incident light. The charged particles may be configured to pass through when the charged particles accumulate on the electrode. Alternatively, the charged particles may be accumulated on the electrode selected by the voltage difference of the electrodes, and the electrode is horizontal to the transmission direction of the incident light. When the charged particles accumulate, the incident light may pass, and when there is no voltage difference between the electrodes, the incident light may be randomly distributed to block incident light.

According to another aspect of the present invention, in the method of forming a biomolecule or polymer array using the above-described programmable mask, (a) irradiating UV light to a selective region using the programmable mask on a molecule having a protecting group while being fixed to the surface And (b) flowing a solution containing a biomolecule or a polymer monomer desired to be immobilized on the molecule, thereby providing a method of forming a biomolecule or polymer array using a programmable mask.

1 is a conceptual diagram for a photoreaction using a transmission type programmable mask used in the present invention.

2 is a plan view of a transmissive programmable mask used in the present invention.

3 is a conceptual diagram of transistors for adjusting transmittance of pixels constituting a transmissive programmable mask.

4 is a conceptual diagram for the transmission / impermeability of UV light using the particles of the present invention.

FIG. 5 is a conceptual diagram of a method of blocking UV transmission of each pixel constituting a transmissive programmable mask.

6 is a conceptual diagram for a method of obtaining UV transmission of each pixel constituting the transmissive programmable mask.

7 is a conceptual diagram of an array of pixels constituting a transmissive programmable mask.

FIG. 8 is a spectrum of UV transmission with and without multiple charged TiO ₂ suspensions between quartz substrates.

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

11: UV light source 12: Transmissive programmable mask

13: Micro Reactor 14: DNA Synthesizer

15: computer 16: euro

17: cable 18, 41, 51, 61, 74: UV light

21: substrate 22: UV light irradiation area

23: drive circuit IC area 24: electrode pad

31, 56, 66: gate wiring 32: gate driving circuit

33, 57, 67: data wiring 34: data driving circuit

35: capacitor 36: storage electrode

42: area in which UV light passage is controlled 43: area in which UV light is blocked

44: particles 52, 62, 73: charged particles

53, 63, 71: horizontal electrodes 54, 64, 72: vertical electrodes

55, 65: voltages applied to the vertical electrodes 58, 68, 77: transistors

59, 69: voltage applied to gate 60, 70, 78, 79: voltage applied to data

75: top plate 76: bottom plate

The object and various advantages of the present invention will become more apparent from the preferred embodiments described below with reference to the accompanying drawings by those skilled in the art.

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

1 is a conceptual diagram for a photoreaction using a transmission type programmable mask used in the present invention. The UV light 18 from the UV light source 11 passes through the transmissive programmable mask 12, the area of passage of which is determined by the transmission of the signal programmed by the computer 15 through the cable 17. The UV light 18 passing through the programmable mask is irradiated on the array substrate to the micro reactor 13 in which the fluid transfer device (not shown) is connected to the substrate on which the DNA array or the polymer array is formed, and photoreaction occurs. The micro reactor 13 is connected to a DNA synthesizer 14 controlled by the computer 15, through which the reagents necessary for forming the array are supplied via the tube 16 and the washing solution required for washing the substrate.

Figure 2 shows a plan view of a transmissive programmable mask used in the present invention. The UV light irradiation region 22 is formed on a quartz-like substrate 21 having excellent UV light transmittance, and is located at the periphery of the region 22, and a driving circuit for selecting a pixel to transmit light by applying an electrical signal. There is an IC region 23, and there is an electrode pad 24 for connecting this driving circuit with an external IC. The transmission of the pixels is determined from the array data obtained from the computer, and this operation is repeated to obtain a DNA or polymer array having a desired arrangement on the substrate.

3 is a conceptual diagram of transistors for adjusting transmittance of pixels constituting a transmissive programmable mask. As a transistor, a silicon thin film transistor is generally used, and the thin film transistor is parallel with the gate wiring 31, the data wiring 33, and the capacitor 35 of the pixel during the switching ON period of each pixel. The storage electrodes 36 are connected to each other to assist the capacitance of the capacitor 35. The gate wiring 31 and the data wiring 33 are electrically connected to the driving IC. Each gate wiring 31 is sequentially selected, and each time the gate wiring 31 is selected, a switching signal enters the data wiring 33 to cause ON / OFF switching of transmittance of each pixel. After switching for all the pixels, the desired transmission pattern is obtained for the entire array.

Figure 4 is a conceptual diagram for the transmission / impermeability of UV light using the particles according to an embodiment of the present invention. In the programmable mask composed of three pixels in FIG. 4, each pixel includes an area 42 through which UV light 41 is controlled and an area 43 with UV light blocked. In the region 43 where the UV light 41 is blocked, a transistor or the like necessary for controlling light is present. The UV light 41 passes through light when the particles 44 which hinder the progress of light are concentrated on the side wall, and the light is blocked when the particles 44 which hinder the progress of the light are spread. To facilitate control of these particles 44, the particles 44 are charged and present in solution. The charged particles 44 can be driven to the electrodes formed on the sidewalls by electrophoresis or dispersed and present in the solution without applying a voltage. In order to make the control of the particles 44 easier, the particles must have a large number of charges and are preferably present in a suspension fluid. In addition, to effectively block UV light, particles 44 must absorb or reflect UV light. The particles 44 may not be easily dissolved in a solution, or may be made of small particles in a molecular form.

In suspensions consisting of a fluid and a large number of charged particles, it is not desirable to use an aqueous solution or the like that absorbs UV light well since the fluid must pass UV light well. Therefore, it is preferable to use fluorocarbon, chlorocarbon, fluorochlorocarbon, and trichlorofluoroethylene, which mainly pass UV well and have good UV stability. .

Particles with a charge may be used, such as TiO _2, barium sulfate, kaolin, and zinc oxide having a size of 500 to 3000 ~.

In addition, a dispersant may be added, and a dispersant may be used as a charge adjuvent, and a polyhydroxy compound and 3-amino-, including ethyl glycol, polypylene glycol, and the like. Amino alcohol compounds, including 3-amino-1 propanol, triethanolamine, etc .; surface modifiers and charge control agents. This can be added.

On the other hand, if the charged particles are not well dispersed in the hydrophobic solvent, unlike the case of Figure 5 by using a different electrode to charge the particles and can block the UV light so that the light is aligned perpendicular to the traveling direction It can also be configured to block UV light. In this case, a transparent electrode such as indium tin oxide (ITO) may be formed in a transparent direction of UV light to apply a voltage to the electrode to configure charged particles to block UV light. 5 and 6, as shown in FIG. 5, the particles 52 are electrophoretically stacked on the transparent electrode 53 perpendicular to the traveling direction of the UV light 51 to block light, and in FIG. 6. As shown, light is transmitted when the particles 62 are electrophoretically accumulated on the horizontal electrode 63 which is perpendicular to the UV light 61 traveling direction. The voltage of the electrode 54 perpendicular to the horizontal electrode 53 can be adjusted by the transistor.

In order to determine transmission / impermeability by one transistor in one pixel, the voltage of the vertical electrode 54 is made constant, and only the voltage of the horizontal electrode 53 is adjusted by the transistor. The voltage corresponding to half of the maximum voltage is applied to the vertical electrode 54 and the ground voltage or the maximum voltage is applied to the horizontal electrode 53 so that the voltage of the vertical electrode 54 is higher or lower than the voltage of the horizontal electrode 53. The charged particles 52 are moved by electrophoresis. Although the horizontal electrode 53 is illustrated as being disposed below the vertical electrode 54 according to the present embodiment, the horizontal electrode 53, the vertical electrode 54, and the type of charge of the particles are applied to the respective electrodes. Depending on the voltage or the like, it may be disposed above or both of the vertical electrodes 54.

7 is a conceptual diagram of an array of pixels constituting a transmissive programmable mask. Each pixel has a transistor connected to it, and there are two electrodes in the space containing the solution through which the UV light 74 passes, and the charged particles 73 accumulate on the electrode 71 perpendicular to the direction of the light. The charged particles 73 are accumulated on the electrode 72 which is parallel to the direction of light. The upper substrate 75 and the lower substrate 76 are bonded to each other so that the solution does not escape. On the other hand, Fig. 7 shows, for example, the gate voltage 79 when turning on the transistor of the unit pixel and the gate voltage 78 turning off.

On the other hand, as described above, in order to confirm the blocking of UV light using the suspension, the UV spectrum with and without the suspension was obtained. FIG. 8 shows a change in wavelength of a UV region in a state in which a suspension containing 1% of 500-3000 mm 3 TiO ₂ particles in a trichlorofluoroethylene solution to which a dispersant is added is injected between a 0.6 mm thick quartz substrate. The transmittance (%) was measured and the transmittance of only the quartz substrate was compared. In the 340-370 nm region known as the most suitable UV wavelength for DNA array production, the permeability of the sample injected with the suspension is about 0.5% and the permeability of the two quartz plates is about 98-99%, so the contrast ratio (contrast ratio) is It is approximately 150: 1. The higher the amount of suspension, the greater the control ratio.

A method of forming an array of biomolecules or polymers using the above-described programmable mask will be described.

First, when UV light is irradiated to a selective region by using the programmable mask on a molecule having a protecting group while being fixed to the surface, the protecting group is separated and the OH group is exposed. If the solution containing the biomolecule or polymer monomer desired to be fixed is flowed, the biomolecule or polymer monomer is fixed only to the exposed portion of the OH group. Since the monomer of the immobilized polymer has another protecting group, it is possible to fix one more monomer by selectively irradiating with light again, and repeating this operation, the biomolecule or polymer array having the desired arrangement Can make

As described above, according to the present invention, an array of biomolecules or polymers such as DNA can be efficiently produced by controlling the transmission of UV light by electrophoresis.

Compared with the conventional photomask, micromirror array, and LCD, it is easy and inexpensive to form a high density array. Since high contrast ratio can be obtained, high purity biomolecule or polymer array can be manufactured. In addition, when a stepping function exists or a plurality of array patterns exist in one programmable mask, mass production of biomolecules or polymer arrays can be easily performed.

In addition, since the programmable mask can be manufactured in a small size, it can be used as a general-purpose DNA chip maker that can be used in hospitals and laboratories. Therefore, high density DNA chips can be manufactured at low cost.

Claims (9)

In the programmable mask for forming a biomolecule or polymer array having an array of unit pixels and a driver circuit unit for selectively applying a voltage to each of the unit pixels, to adjust the transmittance of incident light for each unit pixel, Each of the unit pixels, A solution comprising charged particles that obstruct the progress of incident light and move by electrophoresis; And And a electrode for applying a voltage to the particles to change the arrangement of the particles to control the transmittance of incident light. The method of claim 1, The solution is a programmable mask for forming a biomolecule or polymer array, characterized in that the suspension (fluid) and a suspension fluid consisting of a plurality of charged particles (fluid). The programmable mask of claim 2, wherein the fluid is fluorocarbon, chlorocarbon, or fluorochlorocarbon. 4. 3. The programmable mask of claim 2, wherein the particles are barium sulfate, kaolin, zinc oxide, or TiO ₂ particles having a size of 500-3000 mm ₃ . The method of claim 1, wherein the solution may further include a dispersant, wherein the dispersing agent is a polyhydroxy compound, 3-amino-, which may include ethyl glycol, polypylene glycol Biomolecules characterized in that they are amino alcohol compounds, surface modifiers or charge control agents, which may include 3-amino-1 propanol, triethanolamine Or a programmable mask for forming a polymer array. The programmable mask of claim 1, wherein the charged particles absorb or reflect UV light to block passage of UV light. The method of claim 1, wherein the charged particles are accumulated in the electrode selected by the voltage difference of the electrodes, the incident light is blocked when the charged particles are stacked on the electrode perpendicular to the incident direction of the incident light, A programmable mask for forming a biomolecule or polymer array, characterized in that the incident light passes through when the charged particles accumulate on the electrode parallel to the transmission direction of the incident light. The method of claim 1, wherein the charged particles are accumulated on the electrode selected by the voltage difference of the electrodes, and when the charged particles are accumulated on the electrode parallel to the transmission direction of the incident light, the incident light is passed through, Programmable mask for forming a biomolecule or polymer array, characterized in that when there is no voltage difference between the electrodes to be randomly distributed to block incident light. In the method of forming a biomolecule or polymer array using a programmable mask according to any one of claims 1 to 8, (a) irradiating UV light to an optional region using said programmable mask to a molecule having a protecting group fixed to a surface; And (b) flowing a solution containing a biomolecule or a polymer monomer desired to be immobilized on the molecule; and forming a biomolecule or polymer array using a programmable mask.