KR20100055758A - Microsystem for cell lysis and wireless micro induction device comprising the same - Google Patents

Abstract

PURPOSE: A cell lysis controlling micro system, a bio chip including thereof, and a contactless micro-induction device are provided to improve the portability of the micro-induction device, and to control a cell lysis with a desired temperature and speed. CONSTITUTION: A cell lysis controlling micro system(100) comprises the following: a heating unit(10) heated by a magnetic field; and a micro channel(20) supplied with fluid including cells(30). The heating unit is composed of a plate unit, and a micro structure with multiple cubic figure shapes protruded on the plate unit. The micro structure has an empty cylinder shape and a multi-angle column shape. The plate unit includes a substrate, a first layer formed with Ti/Cu, nickel and nickel-iron on the substrate, and a second layer formed with metal with the magnetism.

Description

Microstructure for cell mechanism control, non-contact micro induction device comprising the same {Microsystem for cell lysis and Wireless micro induction device comprising the same}

The present invention relates to a microstructure for cell mechanism control, and a non-contact micro induction apparatus including the same, and more particularly, to a microstructure for cell mechanism control including a heating unit and a micro channel, and a non-contact micro induction device including the same.

Cell lysis refers to a process for releasing a cell material by breaking a cell membrane, and is mainly a process of extracting from a cell to separate DNA or RNA at a stage of an amplification process such as PCR.

Cell lysis through cell disruption can be largely divided into mechanical and non-mechanical methods. Mechanical methods include ultrasonic, grinder, pressurized (using French press, etc.), reduced pressure, pulverized, and the like, and non-mechanical methods include chemical, thermal, and enzymatic methods.

For example, in the case of sonication conventionally used by mechanical methods, the cell solution or suspension is placed in a chamber located in an ultrasonic bath and sonicated, which has many disadvantages in cell lysis. First, the energy distribution of the ultrasonic waves is not uniform, resulting in inconsistent results, and secondly, the ultrasonic bath does not concentrate energy in the container, and thus there is a problem that more time is spent to complete the destruction of the cells. .

Among the non-mechanical methods, the chemical method uses, for example, an acid, a base, a detergent, a solvent, a chaotropic substance, and the like, and in particular, a method using a detergent is widely used. Detergents break down the lipid bilayer to release the contents of the cells, lysing membrane proteins, which are most commonly used to lyse animal cells, and many detergents denature proteins. However, there is a problem that a reagent must be added separately for cell lysis, a subsequent removal process is required, a PCR inhibitory action occurs, and it takes a long time. In addition, the enzymatic method uses lysozyme, protease, etc. Thermal methods include freeze-thaw methods, heating methods, osmotic shock methods, and electric shock methods. For example, a method of performing cell lysis by thermally contacting a cell with a hot object such as a hot plate, or repeating a cycle of cooling the temperature to about −70 ° C. and thawing to room temperature is used.

The heat cell lysis method as described above is a method suitable for application to a lab-on-a-chip (LOC) because no reagent is used without being a mechanical method. In particular, freeze-thaw is more efficient than cell heating.

Although it is known that this method requires an expensive rapid cooling device to cool down to -70 ° C and takes a long time. In addition, when using a small amount of sample, such as LOC, moisture in the sample is easily evaporated, so that the sample is dried.

Has a problem. In addition, the protein may be denatured to inhibit PCR. This problem also appears in the method of heating only.

Conventional cell lysis methods require a relatively large energy source compared to miniaturized equipment such as biochips, which is unlikely to make biochips larger in size and less convenient and economical.

In addition, since the amount of energy applied to the cell is fixed, it is difficult to control the lysis of the cell finely and variably, and thus, it is difficult to prevent denaturation when the substance released in the cell is sensitive to the external environment.

The energy source must be connected to the biochip by wire, complicating the system and causing a problem.

The present invention is to solve the conventional problems, the object of the present invention is that the energy source is wirelessly connected to the biochip, the equipment can be miniaturized and the control of the applied energy is easy to be customized according to the type of cells Relates to microstructures for possible cell lysis.

Another object of the invention relates to a non-contact micro induction device comprising the microstructure.

One aspect of the invention, the heating unit is heated by a magnetic field; And a micro-channel for controlling the cellular mechanism, wherein the heating unit is located therein and includes a micro channel to which a fluid including cells is supplied.

Another aspect of the invention relates to a non-contact micro induction device comprising the microstructure and an inductor coil that applies a magnetic field to the microstructure.

Another aspect of the invention relates to a method of regulating (or lysing) a cell's mechanism by applying a magnetic field generated by an induction coil to the microstructure.

The non-contact micro induction device including the microstructure for cell mechanism control according to the present invention is portable without wire connection and can share the power supply if only a magnetic field generator is required without additionally installing a power supply to the biochip. You can also control the cells at the desired temperature and speed.

Hereinafter, the present invention will be described in detail.

1 is a schematic diagram showing a microstructure for cell mechanism regulation according to one embodiment of the present invention. 2 is a front view of a heating unit forming the microstructure of the present invention, and FIG. 3 shows a schematic perspective view of the heating unit.

Referring to FIG. 1, the microstructure 100 for cell mechanism control according to the present invention includes a heating unit 10 and a micro channel 20.

The heating unit 10 is heated by a magnetic field.

The micro channel 20 is the heating unit 10 is located therein is supplied with a fluid containing the cells.

2 and 3, the heating unit 10 may include a plate portion 11 and a plurality of three-dimensionally shaped microstructures 12 protruding from the plate portion.

The microstructure 12 may have a hollow cylinder, a polygonal shape inside.

The size of the microstructure 12 may vary depending on the sample of the micro device, the amount of heat generated and the heating temperature. The microstructure 12 has a thickness of 1 to 10000 μm, preferably 1 to 200 μm, an inner diameter of 0.1 to 10000 μm, preferably 1 to 200 μm, and a height of 0.1 to 1000 μm, preferably 1 to 1000 μm. May be μm.

The microstructure 12 is preferably a hollow structure, because the current flows to the outside to control the contact surface and to increase the surface area to increase the heat generating efficiency.

The plate portion 11 may include a substrate 111, a first layer 112 formed of at least one of Ti / Cu, Ni, and Ni / Fe on the substrate, and a metal having conductive or magnetic properties on the first layer. It may include a second layer 113 formed as.

The microstructure 12 and the second layer 113 is at least one metal selected from the group consisting of nickel, nickel iron (NiFe), gold, silver, copper, platinum, aluminum, iron oxide, chromium oxide, cobalt and ferrite It can be formed as.

The plate portion 11 is formed of at least one resin selected from the group consisting of polydimethylsiloxane (PDMS), epoxy, polyurethane, polycarbonate, polymethyl methacrylate, polystyrene and polyethylene terephthalate on the second layer 113 The third layer 114 may be further included.

The third layer may control the degree of exposure of the microstructure 12 as well as the function of the heat transfer layer and the protective layer.

The thickness of the first layer 11 and the thickness of the microstructure 12 may be the same.

The plate portion 11 may include a filler 115 at a lower portion of the microstructure 12. The filler 115 may be formed in a process of forming the microstructure and then removed. As the filler 115, a photoresist, silicon, or the like may be used, but is not necessarily limited thereto.

The microchannel 20 may be formed of at least one selected from the group consisting of glass, poly-di-methyl-siloxane (PDMS), poly-methyl-math-acrylate (PMMA) polystyrene, and polycarbonate. Can be.

The heating unit 10 may control cell lysis and further cell mechanism of the biochip.

Depending on the electrical conductivity and dielectric constant of the material, different eddy currents may flow due to the magnetic field, resulting in a difference in the amount of heat generated. Also, since the eddy currents are denser at the edges than the center, the difference in the amount of heat generated may vary depending on the microstructure 12. have. Therefore, the cells may be lysed at a desired speed and temperature by adjusting the type, shape, thickness, and the like of the material forming the microstructure 12. In addition, the second layer 113 and the third layer 114 may also affect cell mechanism regulation.

4 is a photograph of a microstructure according to another embodiment of the present invention. Referring to FIG. 4, the shape of the protrusion is a hollow rectangular shape. As shown in FIG. 4, the shape and size of the protrusion may be freely manufactured.

Another embodiment of the invention relates to a method of manufacturing the heating unit. 5 is a schematic view showing a step of manufacturing a heating unit.

Referring to Figure 5, the method of manufacturing the heating unit

(A) forming a cylindrical column with the filling 115 on the glass substrate 111

(b) laminating the first layer 112 for nickel electroplating by DC sputtering

(c) spin coating the polymer solution 116 on the first layer

(d) removing the first layer 112 on the cylinder column using reactive ion etching (RIE) after drying of the polymer solution, where the remainder is protected by the polymer.

(e) removing the layer formed by the polymer solution 116

(f) depositing nickel around the cylindrical columnar surface by the step of laminating a second layer 113 over the first layer 112 layer.

(g) depositing a third layer 114 on the second layer 113 and

(h) removing a portion of the filler 115 in the surface of the third layer 114 and the inside of the cylindrical column by using the RIE, by which the microstructure may protrude above the plate-shaped portion. .

In another aspect the invention relates to a biochip comprising the microstructure 100. The biochip is a biosensor, a DNA microarray, a protein chip, a cell chip, a neuron chip and a bio-insertion chip, a lab-on-lab. a Chip) and the like.

Yet another embodiment of the present invention relates to a non-contact micro induction device comprising the microstructure and an inductor coil for applying a magnetic field to the microstructure.

6 is a schematic diagram of a non-contact micro induction device according to an embodiment of the present invention. Referring to FIG. 6, the non-contact micro induction apparatus includes a micro structure 100 and an induction coil 200.

The induction coil 200 can form a necessary alternating magnetic field with a small power source. There is no particular limitation on the power intensity. The induction coil 200 may use a miniaturized or commercially available inductor to be portable. Although the intensity of the magnetic field generated in the inductor coil is not particularly limited, a high frequency alternating magnetic field is preferable, and the frequency may be 10 kHz-100 MHz, preferably 10-500 KHz.

The non-contact micro induction device according to an embodiment of the present invention may include the biochip and the induct coil.

The non-contact micro induction apparatus may heat or heat the heating unit using a micro process to control or dissolve the cell mechanism. That is, cellular mechanisms can be controlled by adsorbing proteins, DNA, RNA, or cells to nanoparticles or microparticles and heating the surfaces of these particles.

In another aspect, an embodiment of the present invention relates to a method of regulating the mechanism of a cell or lysing a cell by applying a magnetic field generated by the induction coil to the microstructure.

The method includes supplying a cell sample to be lysed to a microchannel equipped with a heating unit; Operating an induction coil to apply a magnetic field to the heating unit.

The temperature of the microstructure heated by the magnetic field may be 30 ~ 350 ℃.

In this case, the frequency of the alternating magnetic field may be 10 kHz-100 MHz, preferably 10-500 KHz. The energy applied to the coil is between 0.1 W and 5000 W.

The method can control the movement of the cells, temperature or kill the cells, and also control the behavior of the cell membrane. The method can also regulate the mechanism of proteins, DNA and RNA in the cell.

Hereinafter, the present invention will be described in detail with reference to Examples, but the features of the present invention are not limited to Examples.

Example 1 Preparation of Heating Unit

SU-8 100 (MicroChem, USA) is applied on the glass substrate at 600㎛ and UV is irradiated through a spherical mask having a diameter of 100㎛ to form a cylinder-shaped column having a height of 100㎛ and 600㎛ It was. Subsequently, the Ti / Cu layer is coated using DC-sputter (601 Sputtering System; CVC Products, Rochester, NY), and the poly-lactic-glycolic-acid (PLGA) is melted at 150 ° C. and applied on the Ti / Cu layer. To form a protective layer.

Next, the Ti / Cu layer on the columnar column was removed using Reactive io etching (Plasma-Therm, St. Petersburg, FL), and the polymer layer was removed using ethyl acetate (Aldrich). . Subsequently, nickel was formed on the nickel layer by about 10-15 mm using an electroplating apparatus (Watt formulation bath (Technic, Cranson, RI)). At this time, the nickel layer applied the side surface of the cylindrical column to form a nickel coated column. Next, PDMS (Sylgard 184, Dow Conring) is applied and cured, and RIE (Plasma-Therm, St. Petersburg, FL) is used to remove a portion of the PDMS and a portion of the SU-8 inside the nickel column to annular nickel. The heating unit was manufactured by exposing only the upper part of the microstructure. The microstructure has a height of 600 μm, a diameter of about 100 μm, and a thickness of 10 μm.

Example 2

Put a thermal paper that changes color at 160 ° C or higher on the heating unit manufactured by Example 1, and use the induction equipment (EasyHeat, Ameritherm) at a intensity of 10W to 500W between 0.1 seconds and 10 seconds to 340. A magnetic field was applied at a frequency of KHz to observe whether the temperature of the heating unit rose after a desired time.

Figure 7 is a photograph showing the thermal paper color change after 2 seconds after placing the thermal paper on the heating unit in Example 2. Referring to FIG. 7, since the color of the thermal paper is changed from white to black, it can be confirmed that the temperature of the structure is 160 degrees Celsius or more.

1 is a schematic diagram showing a microstructure for cell mechanism regulation according to one embodiment of the present invention.

2 is a front view of a heating unit forming the microstructure of the present invention.

3 shows a schematic perspective view of the heating unit.

4 is a photograph of a microstructure according to another embodiment of the present invention.

Figure 5 is a schematic diagram showing the step of manufacturing the heating unit.

6 is a schematic diagram of a non-contact micro induction device according to an embodiment of the present invention.

Figure 7 is a photograph showing the thermal paper color change after 2 seconds after placing the thermal paper on the heating unit in Example 2.

* Description of the main parts of the drawings

100: microstructure

10 heating unit 20 microchannel 30 cell

200: induction coil

Claims (10)

A heating unit heated by a magnetic field; And The microstructure for cell mechanism control, characterized in that the heating unit is located therein and comprises a micro channel to which the fluid containing the cells are supplied. The microstructure of claim 1, wherein the heating unit comprises a plate-like portion and a plurality of three-dimensionally-shaped microstructures formed by protruding from the plate-like portion. The microstructure for cell mechanism control according to claim 2, wherein the microstructures are hollow cylinders or polygonal cylinders. 3. The display device of claim 2, wherein the plate portion comprises a substrate, a first layer formed of at least one of Ti / Cu, nickel, and nickel-iron on the substrate, and a second formed of a metal having conductivity or magnetism on the first layer. Cell structure control microstructure, characterized in that it comprises a layer. The method of claim 3, wherein the plate portion is formed of at least one resin selected from the group consisting of polydimethylsiloxane (PDMS), epoxy, polyurethane, polycarbonate, polymethyl methacrylate, polystyrene and polyethylene terephthalate on the second layer Cell structure control microstructure, characterized in that it further comprises three layers. The method of claim 2 or 4, wherein the microstructure and the second layer are selected from the group consisting of nickel, nickel iron (NiFe), gold, silver, copper, platinum, aluminum, iron oxide, chromium oxide, cobalt and ferrite. Microstructure for cell mechanism control, characterized in that formed of at least one metal. Biochip comprising a microstructure according to any one of claims 1 to 6. A non-contact micro induction device comprising a microstructure according to any one of claims 1 to 6 and an inductor coil for applying a magnetic field to the microstructure. A method for regulating the mechanism of a cell by applying a magnetic field generated by an induction coil to the microstructure according to any one of claims 1 to 6. 10. The method of claim 9, wherein the temperature of the microstructure heated by the magnetic field is 30 ~ 350 ℃.

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