KR100815858B1 - Apparatus and method for magnetically separating biological materials from mixture - Google Patents

Abstract

An apparatus and a method for separating biological materials are provided to identify the state of a biological material mixture solution layer through an upper side of an upper plate portion exposed to the outside conveniently and accurately and increase purity of an isolated bio-material by performing a homogenization process without applying magnetic force to the upper plate portion. An apparatus for separating biological materials comprises: a lower plate portion(121) which has a biological material mixture solution receiving portion(122) for receiving a biological material mixture solution where at least one specific bio-material selected from cells, proteins, DNAs and RNAs with magnetic property by attaching a magnetic carrier such as a magnetic bead thereto is mixed; an upper plate portion(111) which is located at the upper part of the lower plate portion to be faced with each other and absorbs the biological material mixture solution received at the biological material mixture solution receiving portion at the lower side; a magnetic force applying means(113) which is located at the upper side of the upper plate portion to be installed separably from the upper plate portion; and an upper and lower plates interval controlling means which is coupled to the upper plate portion or the lower plate portion to control the interval of the upper and lower plate portions narrower, thereby forming a biological material mixture solution layer by the mixture solution being absorbed into the lower side of the upper plate portion and to control the interval of the upper and lower plate portions wider, thereby allowing other biological materials except the specific biological material moved to the upper plate portion by the magnetic force and the specific biological material moved to the lower plate portion by gravity force to be isolated and located at the lower side of the upper plate portion and the lower plate portion, wherein the magnetic force applying means is installed to not apply the magnetic force with exposing the upper plate portion to the outside when isolated from the upper plate portion. The apparatus further comprises an upper housing opened downside and a lower housing(126) opened upside, wherein the magnetic force applying means is located inside of the upper housing and the upper plate portion is installed to place the magnetic force applying means at the upper side, thereby the upper side of the upper plate portion is exposed when the magnetic force applying means is isolated from the upper plate portion; the lower plate portion is installed at the lower housing and the upper and lower plate portions interval controlling means is installed to be coupled to the lower plate portion; and the intervals of the upper and lower plates are controlled by moving up and down using the interval controlling means while the lower side of the upper housing and the upper side of the lower housing are coupled to each other.

Description

Apparatus and method for magnetically separating biological materials from mixture}

The present invention relates to a device and a method for separating a biological material, and more particularly, any one selected from a specific cell, protein, DNA, RNA attached to a magnetic carrier such as magnetic beads The present invention relates to an apparatus and a method for separating a biomaterial by applying a magnetic force on top of the biomaterial mixture liquid layer in which the biomaterial is mixed.

 In PCT / KR2006 / 001077 (WO2006 / 091060) (name of the invention: APPARATUS AND METHOD FOR MAGNETICALLY SEPARATING CELLS FROM MIXTURE), a layer of cell mixture is formed in which specific cells with magnetic beads are mixed between the upper and lower plates. By applying magnetic force on the upper plate side, the specific cells are placed on the upper plate and at the same time, the remaining cells other than the specific cells are placed on the lower plate by gravity, and then the cell mixture layer is disassembled to separate the necessary cells easily and stably. Doing.

Here, the cell mixture layer is formed by adjusting the gap between the upper plate and the lower plate to contact the lower surface of the upper plate with the cell mixture contained in the convex shape upwardly.

The gap between the upper and lower plates of the cell mixture layer formed above is kept wide so as to ensure the separation of the cells by the magnetic force applied to the specific cells and the gravity applied to the rest other than the specific cells. The distance between the upper plate and the lower plate is adjusted by manipulating the dial in which the position of the layer is formed so as to optimize the cell separation state, and then separating the cells.

This cell separation process, as shown in Figure 1, by forming the interior of the conventional cell separation device 10 in a closed space to minimize the evaporation or temperature change of water around the layer to survive the cells during the separation process It proceeds in a state that maintains an optimal environment. In addition, the upper surface is covered by the magnet to apply a magnetic force on the upper surface.

In addition, as shown in Figures 2 and 3, even when the cell mixture is injected in the same amount in several cell mixture solution receiving portion and the amount of the injected itself may be more or less the upper plate portion to form an optimal layer and The spacing of the bottom plate should be different.

And, as shown in Figure 4, in the conventional cell separation apparatus, simply by replacing the lower plate portion and the corresponding upper plate portion formed with a different number of the cell mixture solution receiving portion, the various conditions under the same temperature or humidity environment Isolation of cells with can be performed at one time.

In this case, the interval between the upper plate portion and the lower plate portion forming the layer is different depending on the number of the cell mixture solution receiving portion. For example, as the number of the cell mixture receptacles increases in the lower plate portion having a predetermined area, the diameter becomes smaller. As shown in FIG. 5, the smaller the diameter of the cell mixture receptacles, the optimal layer for separation. The gap between the upper plate portion and the lower plate portion to form a must also be small.

In this way, even if both the amount of the cell mixture injected into the cell mixture containing portion and the diameter of the cell mixture containing portion of the lower plate are different, the conventional cell separation apparatus operates the same dial with the position where the layer is formed in the state which cannot be identified from the outside. There is a problem to form a cell mixture layer and to control the spacing of the formed layer.

In order to solve this problem, a transparent window is installed around the side on which the layer is formed, and the user can check the state of the layer from the outside through the transparent window.

However, it is very inconvenient to check the floor through the transparent window from the size or the installed position of the transparent window and adjust the distance between the layers because only a considerable concentration can be placed in the position of the user's eye level. The production process is complicated because the transparent window must be installed separately to maintain.

On the other hand, if the remaining cells other than the specific cells are also attached to the upper plate subjected to the cell separation process in the above step will have a bad effect on the experiment of the subsequent step it is necessary to increase the purity of the separated specific cells to the maximum.

To this end, as shown in Figure 6, after removing all the cells other than the specific cells located in the lower plate is separated or replaced with a new lower plate and receiving and receiving only the aqueous buffer solution containing no cells in the lower plate Shaking the specific cell mixture layer formed by adjusting the gap between the upper and lower plates to be narrowed up and down as much as possible so that the remaining cells other than the specific cells in the upper plate fall together as a single cell state as they stick together with the specific cells. A homogenization process is performed.

By the way, in the cell separation apparatus, the homogenization process is performed in a state in which a magnetic force that maintains a magnitude greater than or equal to the gravity acting on a specific cell is continuously applied from an upper surface of the upper plate, and thus, the cell separation apparatus is shown in FIG. 7. Likewise, it becomes difficult to single cellize all of the cell masses held together by magnetic force on the top plate side.

In order to solve the above problems, the present invention can easily and accurately check the state of the biomaterial mixture liquid layer through the upper surface of the upper plate exposed to the outside, and also by performing a homogenization process without a magnetic force applied to the upper plate to separate It is an object of the present invention to provide an apparatus and method for separating biomaterials that can increase the purity of a specific biomaterial.

In order to achieve the above object, the present invention provides a biomaterial mixed solution in which any one specific biomaterial selected from a specific cell, protein, DNA, and RNA is attached to a magnetic carrier such as magnetic beads. A lower plate portion provided on the upper surface of the biological material mixture liquid receiving portion accommodated in a convex shape; An upper plate portion facing the upper portion of the lower plate portion, the upper plate portion allowing the biomaterial mixture solution contained in the biomaterial mixture solution accommodation portion to be adsorbed on the lower surface; Magnetic force applying means located on an upper surface of the upper plate portion and detachably installed from the upper plate portion; And, coupled to the upper plate or lower plate, while adjusting the gap between the upper plate and the lower plate to form a biomaterial mixture liquid layer while being absorbed by the lower surface of the biomaterial mixed solution accommodated in the biomaterial mixed solution receiving portion, The other biomaterials other than the specific biomaterial moved to the upper plate part side by the magnetic force applied through the magnetic force applying means to the upper plate portion of the biomaterial mixed liquid layer formed in the lower plate part by gravity, the biomaterial mixed liquid layer And upper and lower plate spacing adjusting means for widening the gap between the upper plate and the lower plate so that the upper plate and the lower plate are spaced apart from each other while being disassembled. When the upper surface of the upper panel is exposed to the external magnetic force It provides a biological material separating device, characterized in that installed so.

Preferably, further comprising an upper housing opened downward and a lower housing opened upward, wherein the upper housing is provided with an upper plate such that the magnetic force applying means and the magnetic force applying means are located on an upper surface thereof, wherein the magnetic force When the application means is separated from the upper plate portion is formed so that the upper surface of the upper plate portion is exposed to the outside, the lower housing, the upper and lower plate portion adjusting means is provided to be coupled to the lower plate portion and the lower plate portion therein, The lower and upper plates are moved up and down by the upper and lower plate spacing adjusting means in a state where the lower and upper portions of the lower housing and the biological material mixed liquid receiving portions of the upper and lower portions of the upper plate face each other. You may have to adjust the spacing.

Preferably, the front portion further comprises a housing, wherein the housing, the magnetic force applying means, the upper plate portion provided with the magnetic force applying means is located on the upper surface and the lower surface of the upper plate and the biomaterial mixed liquid receiving portion facing each other It is provided with a lower plate portion, the upper and lower plate portion adjusting means is installed to be coupled to the upper plate portion to move the upper plate portion up and down to adjust the distance between the upper plate portion and the lower plate portion, the magnetic force applying means is to be separated from the upper surface of the upper plate portion At this time, the housing may be formed such that the upper surface of the upper plate is exposed to the outside.

In addition, in order to achieve the above object, the present invention is selected from a specific cell, protein, DNA, RNA that is magnetic by attaching a magnetic carrier such as magnetic beads accommodated in the convex form in the biomaterial mixture solution receiving portion of the lower plate The gap between the upper plate and the lower plate is narrowed so as to be adsorbed from the lower surface of the upper plate facing the biomaterial mixed solution containing portion of the lower plate to the biomaterial mixed solution layer. Forming a first step; Applying a magnetic force from the top plate side of the biomaterial mixture liquid layer formed in the first step to move the specific biomaterial to the top plate side of the biomaterial mixture liquid layer while the remaining biomaterials other than the specific biomaterial by gravity also lower plate of the biomaterial mixture liquid layer A second step of being moved to the secondary side; In the second step, the remaining biomaterials other than the specific biomaterial moved to the upper plate side of the biomaterial mixture liquid layer and the specific biomaterial moved to the lower plate side are adjusted to widen the gap between the upper plate and the lower plate, so that the biomaterial mixture liquid layer A third step of disassembling such that the upper and lower surfaces of the upper and lower plates are separated from each other; Remove the remaining biomaterials other than the specific biomaterials separated in the third step and located in the lower plate, or replace them with a new lower plate and inject only the aqueous buffer solution containing no biomaterial into the lower plate, and then receive the upper plate and the lower plate. A fourth step of forming a specific biomaterial mixture liquid layer by narrowing the intervals between the portions; A fifth step of homogenizing the specific biomaterial mixture liquid layer by removing the magnetic force applied to the specific biomaterial mixture liquid layer formed in the fourth step and changing the distance between the upper and lower plate portions a plurality of times; Applying the magnetic force to the top plate side of the specific biomaterial mixture liquid layer homogenized in the fifth step to move the specific biomaterial to the top plate side of the specific biomaterial mixture liquid layer while gravity other remaining biomaterials other than the specific biomaterial specific liquid mixture A sixth step of moving to the bottom plate side of the layer; And, in the sixth step, the specific biomaterial and the remaining biomaterials other than the specific biomaterial moved to the upper plate side and the lower plate side of the specific biomaterial mixture liquid layer to widen the gap between the upper plate and the lower plate, so that the layer is dismantled. It provides a biological material separation method consisting of; seventh step to be separated from each other and the biomaterial mixed solution receiving portion of the lower plate and the lower plate.

According to the separation apparatus of the biological material of the present invention having the above configuration, by separating the magnetic force applying means from the upper plate portion so that the magnetic force is not applied while the upper surface of the upper plate portion is exposed to the outside, the magnetic force applying means located on the upper surface of the upper plate portion. When separating from the top plate, the transparent or semi-transparent top plate exposed to the outside makes it easy and accurate to check whether the gap is set so that the formation of the biomaterial mixture layer and the formed layer are optimally separated without forming a separate window. The magnetic force is not applied to the upper plate when the magnetic force applying means located on the upper surface of the upper plate is separated from the upper plate in the process of further separating the other biological materials other than the specific biomaterial contained in the separated specific biological material. Separation of specific biomaterials by performing a homogenization process without It makes it possible to increase the rate.

Hereinafter, a separation device of a biological material according to embodiments of the present invention will be described in detail with reference to the accompanying drawings, taking as an example cell separation.

FIG. 8 is an exploded perspective view of a cell separation apparatus according to an embodiment of the present invention, and FIG. 9 is a cell mixture solution disposed on the lower plate of the lower body in a state in which the upper body is separated into a magnet part and an upper plate part in FIG. 10 illustrates a state in which the injection is accommodated, and FIG. 10 illustrates a state in which the upper plate portion of the upper body is coupled to the lower plate of the lower body in which the cell mixture is injected and received in FIG. 9, and FIG. 11 is injected and received in the lower plate in FIG. 10. The state of the layer formed according to the amount of the cell mixture and the state in which the interval of the layer is optimally adjusted for separation is shown in comparison with the upper surface of the side and the upper plate, Figure 12 is a magnet portion in the upper plate portion of the upper body in FIG. In the case of binding and cell separation, the state around the upper plate and the lower plate is shown in cross section, and FIG. 13 shows the purity of specific cells remaining in the separated upper plate in FIG. 12. When the buffer solution is injected into the new lower plate to accommodate the upper body and the upper body and the lower body are combined, the magnet part of the upper body is separated from the upper plate and the magnetic force is removed to show the state around the upper plate and the lower plate in cross section. FIG. 13 is a cross-sectional view illustrating a process of homogenizing a specific cell mixed solution layer by repeatedly moving the lower plate up and down in FIG. 13, and FIG. 15 is a magnetic force coupled to the upper plate part by combining a magnet part of the upper body with the homogenized specific cell mixed solution layer in FIG. 14. When the cells are separated by the application of the upper and lower plates around the process of change is shown in cross section.

In the description of the cell separation apparatus 100, 100 ′, 100 ″ according to the embodiments of the present invention, in the embodiment of the PCT / KR2006 / 001077, a bolt-shaped connection part is formed at the bottom to form the bolt-shaped connection part. For example, such a PCT has a lower plate portion pedestal 123 which moves the lower plate portion 121 up and down while moving up and down by the rotation of the lower plate portion pedestal moving dial 124 having a nut-shaped connecting portion. / KR2006 / 001077 is a part different from the above, especially the formation of the upper body including the upper housing to confirm the formation of the layer and the control of the spacing of the formed layer while mainly explaining the separation efficiency of the separated cells, the same part The description of the PCT / KR2006 / 001077 is applied as it is and the description is omitted.

As shown in Figure 8 to 12, in the cell separation device 100 according to an embodiment of the present invention consisting of the upper body 110 and the lower body 120, the magnetic force is applied to the lower body 120, the cell mixture is accommodated Performing a process of separating the cells in a state in which the upper body 110 to be applied is integrally bonded, the same as the PCT / KR2006 / 001077.

However, unlike the PCT / KR2006 / 001077, the upper housing in which the magnet 113 and the upper plate 111 are installed so that the magnet 113 is located on the upper surface is a portion in which the magnet 113 is installed. And the upper plate portion 111 are formed to be separated. In this case, when the magnet 113 is separated from the upper plate portion 111, the upper surface of the upper plate portion 111 is exposed to the outside and no magnetic force is applied thereto.

From this, the upper body 110 is upper body magnet portion 110-1 is fixed to the magnet 113 and the upper plate portion 111 is the upper plate upper portion portion 110-2 is fixed by the upper plate fixing means 114. This is in a detachably coupled state.

In this case, the upper housing 115 is divided into a magnet part upper housing in which the magnet 113 is fixed and the upper plate part upper housing in which the upper plate 111 is fixed by the upper plate fixing means 114.

Therefore, the cell separation device 100 according to an embodiment of the present invention, as shown in FIG. 10, separates the upper magnet portion 110-1 from the upper body portion 110-2. Since the upper surface of the upper portion 111 of the transparent or translucent material is exposed to the outside, unlike the conventional process of separating the cells in a state that can not be identified from the outside, it can be immediately confirmed that the cell mixture layer is formed conveniently do.

In addition, the upper plate portion 111 and the lower plate portion (the upper plate portion 111 and the lower plate portion (in order to obtain an optimum state of separation of cells requiring the formed layer in consideration of the difference in the gap between the upper plate portion and the lower plate forming the layer according to the amount of the cell mixture or the diameter of the cell mixture solution ( When adjusting the distance of 121), as shown in Figure 11, it is easier and more accurate to check the diameter through the upper surface than to check the width height of the layer formed between the upper plate and the lower plate from the side.

That is, the cell separation apparatus 100 according to an embodiment of the present invention is disposed at an appropriate position, and the magnet 112 is separated from the upper plate portion 111 to look at the upper surface of the upper plate portion 111 exposed to the outside layer. It is possible to easily determine whether the formation of the width of the formed layer and appropriately set the guide line to form the optimal layer on the upper surface of the upper portion 111 will be more easily confirmed, in this case the upper plate portion exposed to the outside Since 111 is a transparent or translucent material, there is no need to install a separate window, and since the inside is kept in a sealed state, the production is easy.

Therefore, in the state in which the cell mixture is injected into the cell mixture solution receiving portion 122 of the lower plate 121 located in the lower housing 126 of the lower body 120, the upper magnet portion 110-1 is the upper body portion 110-1. The upper body 110 separated from the upper body upper part 110-2 is covered with the lower body 120, and the lower plate part dial 123 is moved up and down with the lower plate part dial 124 to move the upper and lower parts of the lower plate part 121. By adjusting the upper and lower positions, the user optimally optimizes the cell mixture layer and the width of the formed layer while contacting the cell mixture adsorbing unit 112 of the upper plate 111 while looking at the upper surface of the upper plate 111 exposed to the outside. You can check the setting.

As shown in FIG. 12, the upper magnet portion 110-1 is coupled to the upper upper plate portion 110-2 having the layer space therebetween so that a magnetic force is applied to the upper plate portion 111. The cells are moved to the upper plate 111 and the cell mixture layer is dismantled after a certain period of time, for example, 7 to 15 minutes, so that the remaining cells other than the specific cells are placed on the lower plate 121 by gravity.

Then, in the upper plate 111 and the lower plate 121 of the disassembled cell mixture layer, the remaining cells other than the specific cells and the specific cells are located.

Here, in order to increase the purity of the specific cells remaining in the cell mixture adsorbing unit 112 of the upper plate 111, remove all the mixtures of cells other than the specific cells of the lower plate 121 or replace them with a new lower plate. After receiving and injecting an aqueous solution and covering the lower body 120 with the upper body 110, the lower plate part 121 is moved to form a specific cell mixed solution layer. The specific cell moved from the specific cell mixed solution layer to the upper plate part 111 side. They are still under an environment where magnetic force is applied and at the same time gravity can be clearly separated by gravity acting downwards, thus removing the magnetic force to effectively perform the homogenization process.

In the embodiments of the present invention, the magnet 113 is installed to be detachable from the upper plate 111 at the upper surface of the upper plate 111. As shown in FIG. 13, the upper magnet portion 110 of the upper body 110 is illustrated. -1) is separated from the upper body portion 110-2 so that the applied magnetic force does not reach the upper plate portion 111. From this, the gravity of the specific cell mixture layer is only exerted from the upper plate portion 111 so that the cells move toward the lower plate portion 121.

Then, as shown in FIG. 14, while the magnetic force is removed from the upper plate portion 111, the lower plate portion movement dial 124 is rotated left and right to repeatedly move the lower plate portion 121 upward and downward to allow the upper plate portion ( The specific cell mixed solution layer is homogenized while maintaining the specific cell mixed solution layer while repeatedly changing the distance between the 111 and the lower plate 121 many times, for example, less than 10 times.

In this case, the vibration energy is transmitted to each of the cells in the form of agglomerates, thereby destroying the bonds formed between the cells, so that the remaining cells other than the specific cells attached to the specific cells fall and are all single cell state. And evenly distributed throughout the specific cell mixed liquor layer.

Here, in the case of performing the homogenization while repeating disassembly and formation of the specific cell mixture layer, the vibration energy transmitted to each of the cells increases, so that it is easier to single cell than the case of maintaining the specific cell mixture layer. Can be.

The homogenization process can be confirmed from the upper surface of the upper plate portion 111 exposed to the outside can more smoothly control the size of the transmitted vibration energy.

After performing the homogenization process, as shown in FIG. 15, when the upper magnet part 110-1 is coupled to the upper upper plate part 110-2 again and left for 7 to 15 minutes, the While moving the specific cells to the top plate 111 side of the specific cell mixed solution layer by the magnetic force applied through the magnet 113 from the top plate 111 side of the homogenized specific cell mixed solution layer and remaining cells other than the specific cells by gravity at the same time. They are moved to the lower plate portion 121 side of the specific cell mixture solution layer.

In this state, by rotating the lower plate shift dial 124 to disassemble the specific cell mixture solution layer is separated into the specific cells in the upper plate 111 and the remaining cells other than the specific cells in the lower plate 121, respectively. In addition, the remaining cells other than the specific cells included in the isolated specific cells can be further removed to increase the purity of the specific cells required.

From this, the magnetic force applied when performing the homogenization process does not reach the upper plate portion 111, thereby increasing the efficiency of 1 to 3% as a whole compared to the magnetic force applied from the PCT / KR2006 / 001077 to the upper plate portion 111 Could.

In addition, the homogenization process may separate the upper magnet portion 110-1 from the upper upper plate portion 110-2 before forming a specific cell mixture solution layer.

In this case, the cells in the specific cell mixture of the upper plate 111 are positioned downward by gravity, and the lower plate 121 is moved to contact the specific cell mixture of the upper plate 111 with the buffer solution of the lower plate. Forming a cell mixture layer makes single cellization quick and easy.

16 and 17 illustrate a cell separation apparatus according to other embodiments of the present invention in which the upper body is separated into a magnet part and a top plate part.

The cell separation apparatuses 100 ′ and 100 ″ according to other embodiments of the present invention, unlike the embodiment of the present invention, the upper magnet portion 110-1 to the upper body portion 110-2. It is configured in such a way that it is put into or pulled out, or by rotating to lift or lower the hinge of one side to the center, removing the magnetic force in the state that the upper surface of the upper portion 111 is exposed to the outside or the upper surface of the upper portion 111 The same applies to the application of magnetic force while covering.

Figure 18 shows a cell separation device according to another embodiment of the present invention in which the magnet is separated from the top plate.

In the cell separation apparatus 200 according to another embodiment of the present invention, the upper plate portion 230 is moved by moving the upper plate portion 230 in a state where the upper plate portion 230 and the lower plate portion 250 are installed in one housing 210. ) And the cells are separated while adjusting the distance between the lower plate 250.

Specifically, the cell separation device 200 according to another embodiment of the present invention, as shown in Figure 18, the top plate pedestal 240 and the bottom plate pedestal in the interior of the housing 210 is a portion of the front open 260 is provided so that a movement to the front-back side is possible.

Here, the lower plate portion pedestal 260 is fixedly installed on the lower portion of the housing 210, the upper plate portion pedestal 240 is positioned above the lower plate portion pedestal 260 at a predetermined distance and the housing 210 It is installed to be movable up and down in the interior.

The upper plate portion 230 and the lower plate are positioned to face the cell mixture adsorbing portion of the upper plate portion 230 and the cell mixture solution portion of the lower plate portion 250 with grooves formed in the upper plate supporter 240 and the lower plate supporter 260. The unit 250 is installed.

In addition, a magnet 220 is installed on an upper surface of the upper plate support 240 to face an upper surface of the upper plate 230. The magnet 220 is detachably installed back and forth in the housing 210 to be completely separated from the upper plate 230.

In this case, when the magnet 220 is separated from the upper surface of the upper plate 230, the upper portion of the housing 210 is opened so that the upper surface of the upper plate 230 is exposed to the outside, thereby opening the upper opening 280 of the housing. Form or a transparent material.

In addition, the top plate pedestal 240 is gear-coupled (not shown) with the top plate pedestal movement dial 270 installed on the outer side of the housing 210, by the rotation of the top plate pedestal movement dial 270 It is moved up and down. In this case, when the upper plate support 240 moves up and down, the magnet 220 and the upper plate 230 are also integrally moved.

The present invention is not limited to the above embodiments, and various modifications are possible within the scope of the invention described in the claims, and such modifications are included within the scope of the present invention.

In the cell separation apparatus according to the embodiments of the present invention, the top plate or the bottom plate is moved or the magnet is separated and coupled by the user's direct manipulation, but instead of the user's direct manipulation, a driving means such as a motor and the program are controlled by the program. It is also possible to use the microprocessor to automatically operate the preset contents.

In this case, within the range where the cell mixture layer is maintained at a predetermined time and number of times by the control program, the cells are automatically manipulated so that the gap between the upper and lower plates is narrowly or widely changed so that the remaining cells other than the specific cells located on the upper plate side are moved to gravity. By moving to the lower plate side it is possible to further increase the separation efficiency.

Figure 1 shows a conventional cell separation device combined by covering the lower body to the upper body,

Figure 2 shows an example in which the cell mixture is accommodated in the lower plate of the lower body separated from the upper body in Figure 1,

Figure 3 shows the state of the cell mixture layer according to the amount of the cell mixture solution contained in the lower plate in Figure 2,

Figure 4 shows another example in which the cell mixture is accommodated in the lower plate of the lower body separated from the upper body in Figure 1,

Figure 5 shows the state of the cell mixture layer according to the diameter of the cell mixture contained in the lower plate in Figure 4,

FIG. 6 illustrates a state in which an aqueous buffer solution is injected and received in the lower plate to increase the purity of specific cells remaining in the separated upper plate when the upper body is separated from the lower body after performing the cell separation process in FIG. 1.

FIG. 7 is a cross-sectional view illustrating a process of homogenizing in a state in which magnetic force is applied to a specific cell mixed solution layer formed by covering the lower body with the upper body in FIG. 6,

8 is an exploded perspective view of a cell separation device according to an embodiment of the present invention,

FIG. 9 illustrates a state in which the cell mixture is injected and accommodated in the lower plate of the lower body while the upper body is separated into a magnet part and an upper plate part in FIG.

FIG. 10 illustrates a state in which the upper plate part of the upper body is coupled to the lower plate of the lower body in which the cell mixture is injected and received in FIG. 9;

FIG. 11 is a view illustrating a state in which layers are optimally adjusted for separation and a state of layers formed according to the amount of the cell mixture injected and injected into the lower plate in FIG. 10 from the side and the upper plate,

12 is a cross-sectional view showing a state around the upper plate and the lower plate in the case of separating the cells by combining the magnet portion to the upper plate portion of the upper body in Figure 8,

FIG. 13 is a case where the magnetic solution of the upper body is separated from the upper plate by removing the magnetic force after combining the upper body and the lower body by injecting and receiving a buffer aqueous solution into the new lower plate to increase the purity of specific cells remaining in the upper plate separated in FIG. 12. The state around the upper plate and the lower plate is shown in cross section,

FIG. 14 is a cross-sectional view illustrating a process of homogenizing a specific cell mixed solution layer by repeatedly moving the lower plate part up and down in FIG. 13.

FIG. 15 is a cross-sectional view illustrating a process in which the upper and lower plate periphery are changed when the magnet part of the upper body is combined with the upper plate part in the homogenized specific cell mixed solution layer in FIG.

16 and 17 illustrate a cell separation apparatus according to other embodiments of the present invention in which the upper body is separated into a magnet part and a top plate part.

Figure 18 shows a cell separation device according to another embodiment of the present invention in which the magnet is separated from the top plate.

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

100, 100 ', 100 ": cell separation apparatus according to embodiments of the present invention

110: upper body 110-1: upper body magnet portion

110-2: upper body portion 111, 230: upper portion

112: cell mixture adsorbing portion 113, 220: magnet

114: upper plate fixing means 115: upper housing

120: lower body 121, 250: lower plate

122: cell mixture solution receiving portion 123: lower plate support

124: lower plate moving dial 126: lower housing

200: cell separation apparatus according to another embodiment of the present invention

210: housing 240: top plate support

260: lower plate support 270: upper plate support movement dial

280: upper opening of the housing 41: specific cells

42: Magnetic Bead

43: remaining cells other than specific cells

Claims (4)

The upper surface of the biomaterial mixture receiving portion in which a biomaterial mixture containing a specific biomaterial selected from a specific cell, protein, DNA, or RNA, which is attached to a magnetic carrier such as magnetic beads, is received in a convex shape. A lower plate portion provided at the; An upper plate portion facing the upper portion of the lower plate portion, the upper plate portion allowing the biomaterial mixture solution contained in the biomaterial mixture solution accommodation portion to be adsorbed on the lower surface; Magnetic force applying means located on an upper surface of the upper plate portion and detachably installed from the upper plate portion; And, Coupled to the upper plate or lower plate, while adjusting the gap between the upper plate and the lower plate so as to form a biomaterial mixture liquid layer is adsorbed on the lower surface of the upper material of the biological material mixed solution accommodated in the biological material mixture receiving portion, the formed The biomaterial mixture solution layer is dismantled from the other biomaterials other than the specific biomaterial moved to the upper plate part side by the magnetic force applied through the magnetic force applying means to the upper plate portion of the biomaterial mixture liquid layer and to the lower plate part side by gravity. And upper and lower plate spacing adjusting means for widening the gap between the upper plate and the lower plate so that the upper plate and the lower plate are spaced apart from each other in the biomaterial mixture solution receiving portion of the upper plate and the lower plate. The magnetic force applying means, when separated from the upper plate portion is exposed to the outside of the upper plate portion of the biological material separation device, characterized in that the magnetic force is not applied. The method of claim 1, Further comprising a lower housing opened upwards and a lower housing opened upwards, The upper housing is provided with an upper plate portion in which the magnetic force applying means and the magnetic force applying means are positioned on an upper surface thereof, and the upper surface of the upper plate portion is formed to be exposed to the outside when the magnetic force applying means is separated from the upper plate portion. The lower housing, the upper and lower plate space adjusting means is installed to be coupled to the lower plate and the lower plate therein, In the state where the lower surface of the upper plate and the lower portion of the lower plate of the biomaterial mixed liquid receiving portion is coupled to face each other and the upper side of the upper housing and the lower housing are coupled, the upper plate and the lower plate of the upper plate and the lower plate Biomaterial separation apparatus, characterized in that for adjusting the interval. The method of claim 1, The front portion further comprises an open housing, The housing has an upper plate portion provided with the magnetic force applying means, the magnetic force applying means disposed on an upper surface thereof, and a lower plate portion provided with a lower surface of the upper plate portion facing the biomaterial mixed solution receiving portion, The upper and lower plate spacing adjusting means is installed to be coupled to the upper plate portion to move the upper plate up and down to adjust the gap between the upper plate and the lower plate, And the housing is formed such that the upper surface of the upper plate is exposed to the outside when the magnetic force applying means is separated from the upper surface of the upper plate. Biomaterial mixture solution in which any one specific biomaterial selected from specific cells, proteins, DNA, and RNA is mixed with magnetic carriers such as magnetic beads, which are convexly received in the biomaterial mixture receiving portion of the lower plate. A first step of narrowing the gap between the upper plate and the lower plate so as to be adsorbed on the lower surface of the upper plate facing the biomaterial mixed solution receiving portion of the lower plate; Applying a magnetic force from the top plate side of the biomaterial mixture liquid layer formed in the first step to move the specific biomaterial to the top plate side of the biomaterial mixture liquid layer while the remaining biomaterials other than the specific biomaterial by gravity also lower plate of the biomaterial mixture liquid layer A second step of being moved to the secondary side; In the second step, the remaining biomaterials other than the specific biomaterial moved to the upper plate side of the biomaterial mixture liquid layer and the specific biomaterial moved to the lower plate side are adjusted to widen the gap between the upper plate and the lower plate, so that the biomaterial mixture liquid layer A third step of disassembling such that the upper and lower surfaces of the upper and lower plates are separated from each other; Remove the remaining biomaterials other than the specific biomaterials separated in the third step and located in the lower plate, or replace them with a new lower plate and inject only the aqueous buffer solution containing no biomaterial into the lower plate, and then receive the upper plate and the lower plate. A fourth step of forming a specific biomaterial mixture liquid layer by narrowing the intervals between the portions; A fifth step of homogenizing the specific biomaterial mixture liquid layer by removing the magnetic force applied to the specific biomaterial mixture liquid layer formed in the fourth step and changing the distance between the upper and lower plate portions a plurality of times; Applying the magnetic force to the top plate side of the specific biomaterial mixture liquid layer homogenized in the fifth step to move the specific biomaterial to the top plate side of the specific biomaterial mixture liquid layer while gravity other remaining biomaterials other than the specific biomaterial specific liquid mixture A sixth step of moving to the bottom plate side of the layer; And, In the sixth step, the specific biomaterial and the remaining biomaterials other than the specific biomaterial, which are moved to the upper plate side and the lower plate side of the specific biomaterial mixture layer, widen the gap between the upper plate and the lower plate so that the layer is disassembled and the lower surface of the upper plate And a seventh step of separating and respectively placing the biomaterial mixture solution receiving portion of the lower plate portion.

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