KR100862368B1 - Apparatus for applying magnetic field, method for applying magnetic field and apparatus for increasing magnetic gradient therefor - Google Patents

Abstract

An apparatus for applying a magnetic field, a method for applying a magnetic field, and an apparatus for increasing magnetic gradient therefor are provided to move magnetic particles rapidly and to maintain the magnetic particles stably. An apparatus for applying a magnetic field includes a cylindrical body(100), a core(140), supporters(144), a coil, and a power supply. The cylindrical body has a hollow for receiving a well plate(150) having wells(152) therein. The core is inserted in the hollow for fixing the well plate inside the hollow. The supporters are protruded upward from the bottom of the cylindrical body and support the well plate. The coil forms magnetic field around the well plate. The power supply supplies the coil with power.

Description

Magnetic field applying device, magnetic field applying method, and magnetic field gradient increasing device for the same

FIG. 1 is a view showing a result of a magnetic particle moving in a wall plate when a magnetic field is applied to a well plate by a conventional permanent magnet type magnetic field applying device.

FIG. 2A is a diagram showing a conventional permanent magnet type magnetic field applying device, in which permanent magnets are inserted into all holes of the applying device.

FIG. 2B is a view illustrating a conventional permanent magnet type magnetic field applying device, in which a permanent magnet is inserted into some of the holes of the applying device.

Figure 3a is an exploded perspective view of a magnetic field applying apparatus according to an embodiment of the present invention.

FIG. 3B is a cross-sectional view of the assembled application device taken along line AA ′ of FIG. 3A after assembling the components of the magnetic field application device shown in FIG. 3A.

4A is an exploded perspective view of a magnetic field applying apparatus according to another embodiment of the present invention.

4B is a cross-sectional view of the assembled application device taken along the line AA ′ of FIG. 4A after assembling the components of the magnetic field application device shown in FIG. 4A.

5 is a view showing a cooling device mounted around the coil of the magnetic field applying device of the present invention.

6 is a perspective view showing a coil of the magnetic field applying device of the present invention.

FIG. 7A is a perspective view of a magnetic field gradient increasing device in which a plurality of mountain-shaped pillars having short lengths protrude from a disc-shaped plate member. FIG.

FIG. 7B is a front view of the magnetic field gradient increasing device in which a plurality of short-length pillar-shaped pillars protrude from the disc-shaped plate member.

8A is a perspective view of a magnetic field gradient increasing device in which a plurality of short cylinders are formed to protrude from the disk-like plate member.

Fig. 8B is a front view of the magnetic field gradient increasing device in which a plurality of short cylinders protrude from the disc-shaped plate member.

FIG. 9A is a perspective view illustrating a state in which a plurality of long cylinders (3 mm in diameter) are coupled upwardly from a disk-shaped plate member and a structure in which the extended cylinders are fixed so as not to be opened.

9B is a front view showing a state in which a plurality of elongated cylinders (3 mm in diameter) are coupled upwardly from a disk-shaped plate member to form a magnetic gradient increasing device and structures for fixing the extended cylinders so as not to open.

FIG. 9C is a perspective view of one of the structures 147 and 148 separated from each other 148 to fix the cylinders shown in FIGS. 9A and 9B without opening them.

10A is a perspective view illustrating a state in which a plurality of long cylinders (6 mm in diameter) are coupled upwardly from a disk-shaped plate member to form a magnetic gradient increasing device and a structure for fixing the extended cylinders so as not to open.

FIG. 10B is a front view showing a state in which a plurality of long cylinders (6 mm in diameter) are formed to extend upward from a disk-shaped plate member and a structure for fixing the extended cylinders so as not to be opened.

FIG. 11 is a photograph showing a distribution of magnetic particles moved when a magnetic field is applied to magnetic particles in a well plate using a conventional magnetic field applying device and the magnetic field gradient increasing device shown in FIGS. 7A and 7B.

FIG. 12 is a distribution of magnetic particles shifted when a magnetic field is applied to magnetic particles in a well plate by using the magnetic field gradient increasing device shown in FIGS. 7A and 7B and the magnetic field gradient increasing device shown in FIGS. 8A and 8B. Is a picture showing.

FIG. 13 is a photograph showing distribution of magnetic particles shifted when a magnetic field is applied to magnetic particles in a well plate while changing diameters of a plurality of long cylinders in a magnetic field gradient increasing device to 3 mm, 4 mm, and 5 mm.

FIG. 14 shows the distribution of magnetic particles in a well plate that is moved when a magnetic field is applied in the form of a tip of a plurality of long extensions in the form of a mountain and a magnetic field is applied in the form of a cylinder in the magnetic field gradient increasing device. It is a photograph.

<Explanation of symbols for main parts of drawing>

100: cylinder 102: center hole

120: coil 130: power supply

140: core 142: insertion hole

144: a plurality of extensions 146: plate member

147, 148: cylinder fixing structure

150: well plate 152: well

170: fluid

172 hub 174 pipe

180: fluid circulation device 190: moving means

192: lifting shaft 194: lifting guide

1000a, 1000b: magnetic field applying device

200a, 200b, 200c, 200d: magnetic field gradient increasing device

BACKGROUND OF THE INVENTION Field of the Invention The present invention relates to a magnetic field applying device, a magnetic field applying method, and a magnetic field gradient increasing device therefor, and in particular, to reliably increase the magnetic gradients of magnetic particles used in chemical, biological, and biochemical experiments. And to provide a stable. In detail, the present invention provides a magnetic field applying device, a magnetic field applying method, and a method for enabling the magnetic particles to be quickly moved and stably maintained in the moved magnetic particles in chemical, biological, and biochemical experiments using the magnetic particles. A magnetic field gradient increasing device.

In recent years, magnetic particles have attracted much attention in the fields of chemistry, biology, and biochemistry. Magnetic particles can be introduced in a variety of fields because they can introduce a variety of ligands. For example, magnetic particles may be used as carriers for binding proteins, enzymes, drugs, and bioactive molecules, and materials bound to the magnetic particles may be used directly in the course of the experiment or to capture or modify the target protein. ligand).

However, in chemical, biological, and biochemical experiments using magnetic particles, a technique for rapidly moving the magnetic particles to a desired position and stably maintaining the moved magnetic particles is required. In addition, when a bioactive material bound to a magnetic particle reacts with another bioactive material to form a complex, a technique for rapidly moving the complex and stably maintaining the transferred complex is required.

To this end, conventionally, a permanent magnet is installed on a well plate on which a sample including magnetic particles is placed to apply a magnetic field. An example of this is shown in FIGS. 1, 2A and 2B.

In the prior art of FIG. 1, a magnetic field applying device having a permanent magnet formed at a position corresponding to each of the six wells 12 of the well plate 10 is disposed at the bottom of the well plate to apply a magnetic field. However, as shown in FIG. 1, when the magnetic field is applied by the conventional magnetic field applying device, it can be seen that the magnetic particles 14 in the well move to the edge of the well without moving uniformly to the bottom of the well. have.

In addition, a magnetic diffusion plate 40 of the type shown in Figs. 2A and 2B is used as a conventional magnetic field applying device. The self-diffusion plate 40 is formed with a plurality of through-holes 42 in which permanent magnets are inserted to facilitate the arrangement of a plurality of permanent magnets 20, and a plurality of permanent magnets 20 are fixed. By installing and fixing the plurality of through-holes 42 at intervals, the magnetic field generated from the permanent magnet 20 is more uniformly spread.

However, in the prior art as described above, it is impossible to form a magnetic field of uniform intensity in the entire area of the well plate, and the intensity of the magnetic field generated from the permanent magnet is always constant, so it is necessary to adjust the strength of the magnetic field as necessary It was impossible. That is, as a result of moving the magnetic particles and / or magnetic particle complexes (composite of magnetic particles and bioactive materials) in the well plate using the conventional technology, as shown in FIG. The phenomenon occurs that the internal parts are unevenly displaced. Thus, good chemical, biological and biochemical experiments could not be performed.

As such, a number of devices and methods have been proposed for moving magnetic particles and / or magnetic particle complexes in well plates in chemical, biological, and biochemical experiments, but devices for moving magnetic particles easily, uniformly, quickly and stably. The need for a method continues.

In response to this technical need, the present inventors have developed a magnetic field applying device having a new configuration. The new magnetic field applying device is formed by receiving a well plate in a cylinder operating in an electromagnet manner and then supplying power to a coil wound around the cylinder to form a uniform magnetic field in a wide area including the well plate. An example of such a device is disclosed in Korean Patent Application No. 10-2006-119581 (filed November 30, 2006). Nevertheless, as with any good technology, constant improvements can be made.

The force the magnetic particle receives in the magnetic field is not a function of the magnetic field, but a function of the magnetic gradient of the magnetic field. Therefore, when the magnetic field is made uniform, the magnetic particles in the magnetic field receive only the torque associated with the rotation without being pushed in either direction. Therefore, the magnetic particles must have a gradient of the surrounding magnetic field to move under the force from the magnetic field. Therefore, in chemical, biological, and biochemical experiments using magnetic particles, it is not enough to simply increase the strength of the magnetic field and maintain the gradient of the magnetic field in order to move the magnetic particles to the desired position quickly and to keep the moved magnetic particles stable. Augmenting skills are required. However, the new magnetic field applying apparatus described above focuses on uniformly applying the magnetic field to the magnetic particles and controls only the strength of the magnetic field, which takes a long time for the magnetic particles in the well plate to move and stably maintains the moved magnetic particles. The power to let is weak.

Accordingly, the present inventors have come to devise a magnetic field applying device and an application method for providing an increased magnetic field gradient to the magnetic particles in the well plate to quickly move the magnetic particles, and stably maintain the moved magnetic particles. .

The present invention has been made to solve the above problems, and an object of the present invention is to reliably and stably provide an enhanced magnetic field gradient to the magnetic particles used in chemical, biological, and biochemical experiments.

Specifically, the present invention provides a magnetic field applying device, a magnetic field applying method, and an increase in magnetic field gradient for chemical particles, chemical particles, biological particles, and biochemical experiments using magnetic particles to rapidly move magnetic particles and stably maintain the moved magnetic particles. The purpose is to provide a device.

The magnetic field applying device of the present invention for achieving the above object, a plurality of cylinders for accommodating the reaction vessel, and extending in an upward direction with respect to the bottom surface of the cylinder, for increasing the magnetic field gradient for supporting the reaction vessel And an extension, a coil wound around the cylinder to form a magnetic field in a region including the reaction vessel when the power is supplied, and a power supply device for supplying power to the coil.

In one embodiment of the present invention, the central region of the cylinder is made of a non-magnetic magnetic material, a core for fixing the reaction vessel may be installed separately. The plurality of extension parts are formed integrally with the bottom surface of the core. The magnetic field applying device of the present invention can increase the gradient of the magnetic field applied to the reaction vessel by a plurality of extensions integrally formed on the bottom surface of the core. The core may be made of metal such as pure iron, low carbon steel or nickel, and the extension part may be made of metal such as pure iron, cobalt or nickel.

In a preferred embodiment of the present invention, further comprising a plate-like member accommodated in the cylinder, wherein the plurality of extensions extend upwards from the plate-like member with respect to the bottom surface of the cylinder to support the reaction vessel do. The magnetic field applying device of the present invention can increase the gradient of the magnetic field applied to the reaction vessel by a plurality of extension portions extending upward from the plate member with respect to the bottom surface of the cylinder. The plate member is preferably in the form of a thin disk. The plate member and the extension part may be made of metal such as pure iron, cobalt or nickel.

Preferably, the plurality of extension portions may be cylinders, or may be columnar pillars having pointed ends. The diameter of the plurality of extensions is preferably about 0.1mm to 6mm. More preferably, the diameter of the plurality of extensions is about 1 mm to 3 mm. The length of the plurality of extensions is preferably about 25 cm or less. Short mountain extensions and short cylindrical extensions preferably have a minimum length of 0.5 cm.

In one embodiment of the present invention, the reaction vessel is preferably a well plate, it will be appreciated by those skilled in the art that the present invention is not limited thereto. That is, slide glass, vials, or dishes may be used as the reaction vessel, and may be in any form as long as the vessel is subjected to chemical, biological, or biochemical reactions.

In one embodiment of the present invention, copper is generally used as the material of the coil wound around the cylinder, but aluminum may also be used. In one embodiment of the present invention, the coil is wound in a tubular or threaded shape with a good conductive wire such as copper or aluminum. The number of turns of the coil (the number of turns) can be selected from several times to several hundred thousand times according to the purpose of use and the strength of the magnetic field to be applied.

In this invention, when setting the position which becomes 1/2 with respect to the whole height of the said coil as a center position, it is preferable that the position of the reaction container accommodated in the said cylinder is located above from the said center position. In a preferred embodiment of the present invention, the cylinder may be filled with pure iron from its bottom portion to the center position. This can enhance the strength of the applied magnetic field.

In the present invention, the center position of the coil is a region where the strength of the magnetic field formed by the coil is the strongest, but the forces applied to the magnetic particles by the magnetic field in the vertical direction with respect to the center position cancel each other, the magnetic particles are When it is located at the center position is not moved. Therefore, in the present invention, the tip portions of the plurality of extensions for supporting the well plate and increasing the magnetic field gradient in order to achieve the experimental purpose of rapid movement of the magnetic plate to the well bottom of the magnetic plate are spaced upwardly from the center position. It must be located (see Figures 3b and 4b). In addition, as the distance from the center position further upward, the intensity of the magnetic field applied to the well plate is gradually reduced.

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In addition, the magnetic field applying method of the present invention comprises the steps of placing a permanent magnet in the reaction vessel containing a magnetic particle, applying a magnetic field to the magnetic particle by the permanent magnet, and the reaction vessel from the permanent magnet And removing the position, placing the reaction vessel in an electromagnet type magnetic field applying device, and applying a magnetic field having an increased magnetic field gradient by the magnetic field applying device to the magnetic particles.

The magnetic field gradient increasing device of the present invention includes a plate-like member accommodated in the cylinder of the magnetic field applying device, and a plurality of extension portions extending upward from the plate-like member with respect to the bottom surface of the cylinder and supporting the reaction vessel.

The plate member is preferably in the form of a thin disk. The plate member and the extension part may be made of metal such as pure iron, cobalt or nickel.

Hereinafter, the present invention will be described in more detail based on examples. The following examples of the present invention are not intended to limit or limit the scope of the present invention only to embody the present invention. From the detailed description and examples of the present invention, those skilled in the art to which the present invention pertains can easily be interpreted as belonging to the scope of the present invention. References cited in the present invention (Korean Patent Application No. 10-2006-119581) are incorporated herein by reference.

First, referring to FIGS. 3A to 6, the magnetic field applying apparatuses 1000a and 1000b of the present invention may include a cylinder 100 for accommodating a well plate 150 having a plurality of wells 152 and a cylinder 100. Protruding upward with respect to the bottom surface of the plurality of extensions 144 supporting the well plate 150, and winding several to hundreds of thousands of times around the cylinder 100 to supply the well plate 150 when power is supplied. Coil 120 for forming a magnetic field in the region containing, and a power supply device 130 for supplying power to the coil 120.

The coil 120 is wound several times to hundreds of thousands of times the copper of the wire rod in a tubular shape. The number of windings of the coil 120 may be selectively varied according to the strength of the magnetic field to be applied.

The central region of the cylinder 100 is made of a nonmagnetic magnetic material, and installs a core 140 for fixing the well plate 150. The core 140 is made of metal such as pure iron, low carbon steel or nickel. When the core 140 is magnetized, the well plate 150 is attracted to the core 140 and thus may interfere with a desired biological experiment. Therefore, the core 140 is formed of a nonmagnetic magnetic material that can be magnetized only while a current flows. This is preferred. By installing the core 140, a phenomenon in which the magnetic field is concentrated toward the core 140 occurs, thereby increasing the strength of the magnetic field near the core 140.

In the magnetic field applying apparatus 1000a illustrated in FIGS. 3A and 3B, the plurality of extension portions 144 are integrally formed on the bottom surface of the core 140 as a columnar pillar having a sharp tip. In the magnetic field applying apparatus 1000b shown in FIGS. 4A and 4B, the plurality of extension portions 144 are formed integrally on the bottom surface of the core 140 as a cylinder. The diameter of the plurality of extensions 144 is preferably about 0.1mm to 6mm, more preferably about 1mm to 3mm, but is not limited thereto. In the present invention, the magnetic field gradient is increased by the plurality of extensions 144 so that the magnetic particles in the well 152 of the well plate 150 can be quickly moved to the bottom surface of the well 152, and the moved magnetic particles are moved. It will be able to strengthen the power to hold.

7A to 10B illustrate various types of magnetic field gradient increasing apparatuses 200a, 200b, 200c, and 200d of the present invention.

Typically, the magnetic field gradient increasing apparatuses 200a, 200b, 200c, 200d are plate-like members 146 accommodated in the cylinder 100 of the magnetic field applying device, and from the plate-like member 146 upward with respect to the bottom surface of the cylinder 100. And 96 (8 horizontal x 12 vertical) extensions 144 that project in the direction and support the 96-well plate 150. The magnetic field gradient increasing apparatuses 200a, 200b, 200c, and 200d are inserted into the core 140 of the magnetic field applying apparatuses 1000a and 1000b shown in FIGS. 3A to 4B and additionally installed, or for increasing the magnetic field gradient. The extension portion 144 may be inserted into the cylinder or the core of the magnetic field applying device, which is not provided, to increase the magnetic field gradient.

The plate member 146 may be in the form of a thin disk, and the plate member 146 and the extension 144 may be made of metal such as pure iron, cobalt or nickel. The plate member 146 and the extension part 144 may be integrally formed as a casting, or may be manufactured separately, and various manufacturing methods for assembling the extension part 144 to the plate member 146 may be used. In addition, the embodiment of the present invention has been described as an example of the magnetic field gradient increasing device formed with 96 extensions, the present invention is not limited thereto. That is, those skilled in the art will readily understand that the number and arrangement of extensions may be appropriately adjusted to the number and arrangement of wells in the well plate.

7A and 7B, a magnetic field gradient increasing device 200a is formed in which a plurality of short-length mountain pillars 144 protrude from the disk-shaped plate member 146. The device 200a is inserted into the cylinder 100 or the core 140 of the magnetic field applying device, the well plate 150 is placed on the magnetic field gradient increasing device 200a, and then the coil 120 of the magnetic field applying device. When a current is applied to the magnetic field, the magnetic field having the increased magnetic gradient is applied to the magnetic particles in the well plate 150.

8A and 8B illustrate a magnetic field gradient increasing device 200b in which a plurality of short cylinders 144 protrude from the disk-shaped plate member 146. It has been found that the magnetic field gradient increasing device 200b having the extension portions 144 of the short cylinders is more efficient than the magnetic field gradient increasing device 200a having the extension portions 144 of the pillars in the form of short mountains.

9A and 9B show a magnetic field gradient increasing device 200c formed by extending long cylinders 144 having a diameter of 3 mm upward from the disk-shaped plate member 146 and the extended cylinders 144 so as not to be opened. Fixing structures 147 and 148 show a combined state. Among the structures, the disk-shaped structure 147 is a structure for fixing the extensions 144 on the top in consideration of a problem that long cylinders may be extended or overlap each other, and is made of a plastic material. However, the present invention is not limited thereto, and any material that is not magnetic can be applied to the disk-shaped structure 147. Holes corresponding to the long cylinders are formed in the disc-shaped structure 147, and long cylinders are inserted into the holes to prevent long cylinders from being opened. On the other hand, another structure 148 shown in the separated state is also formed in the holes corresponding to the long cylinders, the long cylinders are inserted one by one into these holes is securely fixed without being shaken.

10A and 10B show a magnetic field gradient increasing device 200d formed by extending the long cylinders 144 having a diameter of 6 mm upward from the disk-shaped plate member 146 and the extended cylinders 144 so as not to be opened. The fixing structure 147 is shown in a coupled state. The disk-shaped structure 147 is a structure for fixing the extension parts 144 from the top in consideration of the problem that the elongated cylinders can be opened or overlap each other, it is made of a plastic material. However, the present invention is not limited thereto, and any material that is not magnetic can be applied to the disk-shaped structure 147. Holes corresponding to the long cylinders are formed in the disc-shaped structure 147, and long cylinders are inserted into the holes to prevent long cylinders from being opened.

In the present invention, the core 140 is formed in a cylindrical shape in which the upper end is open, but the present invention is not limited thereto and the core may be configured in various forms. An insertion hole 142 into which the well plate 150 is inserted is formed at the center of the core 140. Therefore, since the upper end of the core 140 is open, the well plate 150 can be put in and taken out from time to time for the purpose of experimentation, and thus, chemical, biological, and biochemical experiments can be performed while maintaining a continuously applied magnetic field. .

The well plate 150 is fixed at a predetermined interval upward from a region where the strength of the magnetic field formed by the coil 120 is the strongest, that is, the central position of the coil. Accordingly, the magnetic particles in the well 152 of the well plate 150 move to the bottom surface of the well.

In one embodiment of the present invention is provided with a moving means 190 for moving the core 140 in the vertical direction to change the strength of the magnetic field applied to the well plate 150. The moving means 190 includes a lifting shaft 192 and a lifting guide 194 for guiding the lifting shaft 192 up and down. The lifting shaft 192 is directly connected to the bottom surface of the core 140 or is in contact with the bottom surface of the core to transfer the movement in the vertical direction to the core. The lifting shaft 192 formed of a male screw meshes with the lifting guide 194 formed of a female screw and rotates clockwise or counterclockwise to move up and down with respect to the lifting guide. The lifting shaft 192 is driven by driving means such as a hydraulic pump, a solenoid, or a motor. For example, when the rotational force of the driving means is transmitted to the lifting shaft formed by the male screw, the lifting shaft moves upward and downward while rotating with respect to the lifting guide formed by the female screw, thereby appropriately positioning the core 140 connected to one end of the lifting shaft. Will be moved to. Therefore, the moving means 190 may serve to position the well plate 150 fixed to the core 140 above the region having the strongest magnetic field.

On the other hand, in a preferred embodiment of the present invention, in order to enhance the strength of the applied magnetic field, the cylinder 100 is filled with pure iron from its bottom portion to its center position (not shown). In this case, the configuration of the moving means 190 may be omitted.

In addition, in one embodiment of the present invention, the power supply device 130 is provided with a transformer (not shown) for converting a voltage into a current to apply to the coil 120. The current value applied to the coil 120 can be adjusted by this transformer. For example, the transformer may convert a voltage of 380V into a current of 0A to 100A.

As described above, the strength of the magnetic field suitable for chemical, biological, and biochemical experiments for moving the magnetic particles is preferably 1,000 to 20,000 G (Gauss), and more preferably about 3,500 to 10,000 G (Gauss). Therefore, in the present invention, the magnetic field applied to the magnetic particles in the well plate by adjusting the number of windings of the coil, whether the core is installed, the position of the core (adjust the height), whether the pure iron is charged in the cylinder, and the current value applied to the core You can adjust the intensity of the 3,500 ~ 10,000 G (Gauss).

On the other hand, while the current is applied to the coil to generate a magnetic field, heat is generated from the coil 120, the excessive heat generation may shorten the life of various components, may adversely affect the sample in the well 152. Therefore, in one embodiment of the present invention, cooling means for dissipating heat generated from the coil 120 is provided. The cooling means includes a pipe 174 disposed at a position close to the coil 120, a heat transfer fluid 170 embedded in the pipe 174, and a fluid circulation device for circulating the fluid 170 to the outside ( 180). As the fluid 170, water or oil having a good heat transfer rate may be applied. The fluid circulation device circulates the fluid 170 into the plurality of pipes 174 through the supply pipe 171 and the hub 172.

Hereinafter, the operation of the magnetic field applying device having the above configuration will be described.

The 96-well plate 150 including the magnetic particles is extended from the bottom surface of the core 140 through the insertion hole 142 of the core 140 mounted in the center hole 102 of the cylinder 100. It is positioned on 96 protruding portions 144 that protrude. In the case of using the magnetic field gradient increasing apparatuses 200a-200d, first, the magnetic field gradient increasing apparatuses 200a-200d are provided through the insertion hole 142 of the core 140 mounted in the central hole 102 of the cylinder 100. Is placed on the bottom surface of the core 140. Then, the 96-well plate 150 including the magnetic particles is placed on the 96 extension portions 144 protruding from and protruding from the disk-shaped plate members 146 of the magnetic field gradient increasing apparatuses 200a-200d. . In this case, each of the 96 wells 152 of the well plate 150 is positioned to correspond to each of the 96 extensions 144.

Then, the core 140 is moved through the moving means 190 to adjust the position of the core so that the bottom surface of the core 140 is slightly above the center position of the coil. Then, when power is applied to the coil 120 from the power supply device 130, a magnetic field is formed in the region of the central hole 102 of the cylinder 100, which is an inner region of the coil 120, the magnetic field is the center of the coil Since the position is the strongest, all magnetic materials exposed to the magnetic field are attracted to the center position of the coil. In addition, when the magnetic field strength at the distal end of the 96 extension portions 144 of the magnetic field gradient increasing apparatuses 200a to 200d and the magnetic field strength between the distal end and the distal end (hereinafter referred to as the magnetic field strength between distal ends) are measured and compared, The strength of the magnetic field between the distal ends "becomes larger than the" magnetic field strength at the distal end ", so that an increased magnetic field gradient is formed from the position between the distal ends to the distal end of the extension part. Thus, an increased magnetic field gradient is applied to each well 152 positioned correspondingly above the tip of each extension 144. Accordingly, the magnetic particles present in each well 152 quickly move to the bottom surface of the well 152, and the moved magnetic particles can be stably maintained at the bottom surface of the well 152. Therefore, the magnetic particles or the magnetic particle complexes stably maintained at the bottom of the well can be easily separated from the sample.

Example 1

Comparison of Magnetic Particle Movement between Conventional Magnetic Field Applicator and Magnetic Field Applicator of the Present Invention

First, magnetic nanoparticles (MNP) present in each well 152 of the 96-well plate 150 using a conventional magnetic field applying device (Patent Application No. 10-2006-119581) to be compared. ) And within each well 152 of the 96-well plate 150 using a magnetic field application device inserted into the core 140 as shown in FIGS. 7A and 7B. An experiment was performed to move the existing magnetic nanoparticles.

100 ml of superparamagnetic nanoparticles (50 nm) was introduced into each well of a µClear 96-well plate (purchased from Greiner, Cat. No. 655090) to prepare a sample to be measured. Next, in the conventional magnetic field applying device, the 96-well plate 150 is placed on the bottom surface of the core 140 through the insertion hole 142 of the core 140. On the other hand, in the case of using the magnetic field gradient increasing device 200a in which the short mountain extension portions 144 protrude from the disc-shaped plate member 146, the magnetic field is inserted through the insertion hole 142 of the core 140. After positioning the gradient intensifier 200a on the bottom surface of the core 140, a 96-well plate 150 including magnetic particles is extended from the disk-shaped plate member 146 of the magnetic field gradient intensifier 200a. It was placed on 96 protruding portions 144 protruding. In this case, each of the 96 wells 152 of the well plate 150 is positioned to correspond to each of the 96 extensions 144.

Then, the core 140 of the conventional magnetic field applying device or the magnetic field applying device of the present invention is moved through the moving means 190 so that the bottom surface of the core 140 is located slightly upward from the center position of the coil. Adjust the position. Then, a current of 100 A was applied and maintained from the power supply device 130 to the coil 120. At this time, as a result of measuring the strength of the magnetic field on the bottom surface of the core 140, the magnetic field strength of about 10,000 gauss was measured. Divided into seven experimental groups in total, the photographs were taken at the following time intervals after applying the magnetic field to each experimental group (see FIG. 11). The wash was performed only once last. On the other hand, in the experimental group 3) to 6) below, the additional magnetic field using the permanent magnet was performed before and after applying the magnetic field of the electromagnet method to the 96-well plate 150. The magnetic field strength of the permanent magnets used was about 3,000-4,000 gauss.

1) Photographs were taken at time points of 0 minutes, 5 minutes, 15 minutes, 30 minutes, and 1 hour after application of the magnetic field by the conventional magnetic field applying device. Washing was done only once in the last step;

2) Photographs were taken at time points of 0 minutes, 5 minutes, 15 minutes, 30 minutes, and 1 hour after application of the magnetic field using the magnetic field gradient increasing device of the present invention. Washing was done only once in the last step;

3) Photographs were taken at 0, 5, and 15 minutes after application of the magnetic field by the conventional magnetic field application device. Then, the 96-well plate was removed from the magnetic field application device and additional magnetic field application was performed by the permanent magnet. After that, the photographs were taken at time points of 0 minutes, 5 minutes, 15 minutes, 30 minutes, and 1 hour. Washing was done only once in the last step;

4) Photographs were taken at 0, 5, and 15 minutes after application of the magnetic field using the magnetic field gradient increasing device of the present invention. Then, the 96-well plate was removed from the magnetic field applying device and additional magnetic field was generated by the permanent magnet. Photographs were taken at time points of 0, 5, 15, 30 and 1 hour after the application. Washing was done only once in the last step;

5) Photographs were taken at 0, 5, and 15 minutes after the magnetic field was applied by the permanent magnet, and then the 96-well plate was removed from the permanent magnet and applied after the magnetic field was applied by the conventional magnetic field applying device. Photographs were taken at time points of 0 minutes, 5 minutes, 15 minutes, 30 minutes and 1 hour. Washing was done only once in the last step;

6) Photographs were taken at 0, 5, and 15 minutes after the magnetic field was applied by the permanent magnet, and then the 96-well plate was removed from the permanent magnet and the magnetic field using the magnetic field gradient increasing device of the present invention. Photographs were taken at 0, 5, 15, 30 and 1 hour after application. Washing was done only once in the last step;

7) Photographs were taken at 0, 5, 15, 30, 1, 1 and 30 minutes after the magnetic field was applied by the permanent magnet.

As shown in FIG. 11, when only the conventional electromagnetic field applying device is used, magnetic particles are not aggregated at 5 minutes after application of the magnetic field. However, the magnetic field gradient increasing device of the present invention is attached. In this case, it was observed that the magnetic particles agglomerated at 5 minutes after the magnetic field was applied. In addition, it was observed that the magnetic particles were continuously maintained when the 96-well plate was left in the permanent magnet after 15 minutes of application of the magnetic field using the magnetic field gradient increasing device of the present invention. On the other hand, when only permanent magnets were treated in 96-well plates, magnetic particles tended to diffuse when the well plates were moved for further experimentation. In addition, after maintaining the permanent magnet on the 96-well plate for 15 minutes, it can be observed that even when the magnetic field is applied to the 96-well plate by using the magnetic field gradient increasing device of the present invention, the magnetic particles are continuously maintained. there was.

Example 2

Magnetic gradient intensifier with short mountain extensions or short cylindrical extensions

A magnetic field gradient increasing device 200a as shown in FIGS. 7A and 7B or a magnetic field gradient increasing device 200b as shown in FIGS. 8A and 8B is installed using a magnetic field applying device inserted into the core 140. An experiment was performed to move the magnetic nanoparticles present in each well 152 of the well plate 150.

100 ml of superparamagnetic nanoparticles (50 nm) was introduced into each well of a µClear 96-well plate (purchased from Greiner, Cat. No. 655090) to prepare a sample to be measured. Next, the magnetic field gradient increasing device 200a or the magnetic field gradient increasing device 200b is positioned on the bottom surface of the core 140 through the insertion hole 142 of the core 140. Then, the 96-well plate 150 including the magnetic particles extends from the disc-shaped plate member 146 of the magnetic field gradient increasing device 200a and protrudes in a short hill form. Alternatively, it was placed on 96 extension portions 144 extending from the disk-shaped plate member 146 of the magnetic field gradient increasing device 200b and protruding in a short cylindrical shape. In this case, each of the 96 wells 152 of the well plate 150 is positioned to correspond to each of the 96 extensions 144.

Then, the core 140 of the magnetic field applying device is moved through the moving means 190 to adjust the position of the core so that the bottom surface of the core 140 is slightly above the center position of the coil. Then, a current of 100 A was applied and maintained from the power supply device 130 to the coil 120. At this time, as a result of measuring the strength of the magnetic field on the bottom surface of the core 140, the magnetic field strength of about 11,000 gauss was measured. Divided into seven experimental groups in total, the photographs were taken at the following time intervals after applying magnetic fields in each experimental group (see FIG. 12). The wash was performed only once last. On the other hand, in the experimental group 3) to 6) below, the additional magnetic field using the permanent magnet was performed before and after applying the magnetic field of the electromagnet method to the 96-well plate 150. The magnetic field strength of the permanent magnets used was about 3,000-4,000 gauss.

1) Photographs were taken at time points of 0 minutes, 5 minutes, 15 minutes, 30 minutes, and 1 hour after application of the magnetic field using the magnetic field gradient increasing device 200b. Washing was done only once in the last step;

2) Photographs were taken at time points of 0 minutes, 5 minutes, 15 minutes, 30 minutes, and 1 hour after application of the magnetic field using the magnetic field gradient increasing device 200a. Washing was done only once in the last step;

3) Photographs were taken at 0, 5, and 15 minutes after application of the magnetic field using the magnetic field gradient increasing device 200b. Then, the 96-well plate was removed from the magnetic field applying device and additional magnetic field was applied by the permanent magnet. Photographs were taken at 0, 5, 15, 30 and 1 hour after the operation. Washing was done only once in the last step;

4) Photographs were taken at 0, 5, and 15 minutes after application of the magnetic field using the magnetic field gradient increasing device 200a. Then, the 96-well plate was removed from the magnetic field applying device and additional magnetic field was applied by the permanent magnet. Photographs were taken at 0, 5, 15, 30 and 1 hour after the operation. Washing was done only once in the last step;

5) Photographs were taken at 0, 5, and 15 minutes after the magnetic field was applied by the permanent magnet. Then, the 96-well plate was removed from the permanent magnet and the magnetic field using the magnetic gradient increasing device 200b. Photographs were taken at 0, 5, 15, 30 and 1 hour after application. Washing was done only once in the last step;

6) Photographs were taken at 0, 5, and 15 minutes after the magnetic field was applied by the permanent magnet. Then, the 96-well plate was removed from the permanent magnet and the magnetic field using the magnetic field gradient increasing device 200a. Photographs were taken at 0, 5, 15, 30 and 1 hour after application. Washing was done only once in the last step;

7) Photographs were taken at 0, 5, 15, 30, 1, 1 and 30 minutes after the magnetic field was applied by the permanent magnet. Washing was done only once in the last step.

As shown in FIG. 12, it was observed that the magnetic particles agglomerated at 5 minutes after applying the magnetic field using the magnetic field gradient increasing device 200a or the magnetic field gradient increasing device 200b. In addition, it is observed that when the 96-well plate is left in the permanent magnet after 15 minutes of application of the magnetic field using the magnetic field gradient increasing device 200a or the magnetic field gradient increasing device 200b, the magnetic particles are continuously maintained. Could. On the other hand, when only permanent magnets were treated in 96-well plates, magnetic particles tended to diffuse when the well plates were moved for further experimentation. In addition, after the permanent magnet is maintained on the 96-well plate for 15 minutes, the magnetic particles are agglomerated even when the magnetic field is applied to the 96-well plate using the magnetic field gradient increasing device 200a or the magnetic field gradient increasing device 200b. It was observed that this persisted.

Example 3

Comparison of short cylinder and long cylinder

Experimental substantially the same as Example 2 by changing the cylinder length of the magnetic field gradient increasing device 200c or the magnetic field gradient increasing device 200d as shown in FIGS. 9A to 10C to 25 cm and changing the diameter to 5 mm, 4 mm, and 3 mm. Was performed. However, the point at which 100 µl of superparamagnetic nanoparticles (50 nm) were introduced into each well was different from the photographing time interval.

In the case of applying the magnetic field using the magnetic field gradient increasing device 200b having short cylinders, if the 96-well plate is held in the magnetic field applying device for 15 minutes and then left in the permanent magnet, the aggregation of magnetic particles is continuously maintained. However, when magnetic field is applied using a magnetic field gradient increasing device having long cylinders having a length of 25 cm, agglomeration of magnetic particles is observed from about 10 minutes after application of the magnetic field, but magnetic particles are applied when washed after applying the magnetic field for about 1 hour. Easier separation was observed than in Example 2 (see FIG. 13).

Example 4

Comparison of long mountain columns and long cylinders

Substantially the same experiment as in Example 2 was performed with the extension of the magnetic field gradient increasing device 200c as shown in FIGS. 9A and 9B having a length of 17 cm and the tip thereof being in a mountain form or flattened. However, the point at which 100 µl of superparamagnetic nanoparticles (50 nm) were introduced into each well was different from the photographing time interval.

As shown in FIG. 14, a magnetic field is applied to a 96-well plate using a magnetic field gradient increasing device having a long cylindrical extension (3 mm in diameter) rather than a magnetic field gradient increasing device having a long mountain shaped extension (3 mm in diameter). It was confirmed that the cohesion of the magnetic particles was good in the case of applying.

As described above, the magnetic field applying device, the magnetic field applying method, and the magnetic gradient increasing device therefor can reliably and stably provide an increased magnetic field gradient to magnetic particles used in chemical, biological, and biochemical experiments. .

That is, the present invention has an advantage of enabling the rapid movement of magnetic particles and stably maintaining the moved magnetic particles in chemical, biological and biochemical experiments using magnetic particles.

The present invention has been described above by way of specific embodiments, but the present invention is not limited to the above-described embodiments, and those skilled in the art without departing from the gist of the present invention claimed in the claims. Anyone can make a variety of variations.

Claims (16)

Cylinder for accommodating the reaction vessel, A plurality of extensions extending upwardly with respect to the bottom surface of the cylinder and for increasing a magnetic field gradient supporting the reaction vessel; A coil wound around the cylinder to form a magnetic field in a region including the reaction vessel when power is supplied; Magnetic field applying apparatus comprising a power supply for supplying power to the coil. The method of claim 1, The magnetic field applying device is made of a nonmagnetic magnetic material in the central region of the cylinder further comprises a core for fixing the reaction vessel. The method of claim 2, And the plurality of extension parts are integrally formed on the bottom surface of the core. The method of claim 3, The core is made of pure iron, low carbon steel or nickel, the extension portion of the magnetic field applying device, characterized in that consisting of pure iron, cobalt or nickel. The method according to any one of claims 1 to 4, And a plate member housed in the cylinder, wherein the plurality of extension portions extend upward from the plate member with respect to the bottom surface of the cylinder to support the reaction vessel. The method of claim 5, The plate member is in the form of a thin disk, the plate member and the extension portion magnetic field applying device, characterized in that consisting of pure iron, cobalt or nickel. The method according to any one of claims 1 to 4, The plurality of extension portion is a cylinder, or a magnetic field applying device, characterized in that the end is a pointed column-shaped pillar. The method of claim 7, wherein Magnetic field applying apparatus, characterized in that the diameter of the plurality of extensions is 0.1mm to 6mm. delete delete Placing a permanent magnet in a reaction vessel containing magnetic particles, Applying a magnetic field to the magnetic particles by the permanent magnet; Removing the reaction vessel from the permanent magnet; Positioning the reaction vessel in an electromagnetic field applying device; And applying a magnetic field having an increased magnetic field gradient by the magnetic field applying device to the magnetic particles. Magnetic field including a cylinder for accommodating the reaction vessel, a coil wound around the cylinder to form a magnetic field in the region containing the reaction vessel when the power is supplied, and a power supply for supplying power to the coil. A plate member accommodated in the cylinder in the application device; A magnetic field gradient increasing device comprising a plurality of extensions extending upwardly from the plate member with respect to the bottom surface of the cylinder. The method of claim 12, The plate member is in the form of a thin disk, the plate member and the extension portion magnetic gradient gradient device, characterized in that consisting of pure iron, cobalt or nickel. The method according to claim 12 or 13, The plurality of extension portions are cylindrical or magnetic field gradient increasing device, characterized in that the end is a pointed mountain-shaped pillar. The method of claim 14, Magnetic field gradient increasing device, characterized in that the diameter of the plurality of extensions is 0.1mm to 6mm. delete

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