KR20100070714A

KR20100070714A - Apparatus for separation of samples

Info

Publication number: KR20100070714A
Application number: KR1020080129391A
Authority: KR
Inventors: 박일중
Original assignee: 박일중
Priority date: 2008-12-18
Filing date: 2008-12-18
Publication date: 2010-06-28

Abstract

The present invention relates to a sample separation device, and more particularly, a centrifugation and electrophoresis can be performed at the same time for faster separation of a sample, and at the same time a centrifugal force and an electric force simultaneously act on a particle separation device for finer separation. It is about.

The present invention provides a rotatable disk shaped platform; A gel matrix formed on the upper portion of the platform in the same shape as that of the platform and configured for electrophoresis; A (+) electrode provided at point A, which is any point of the matrix, and a (-) electrode provided at point B, which is point symmetrical with the point A with respect to the center of the matrix; DC power supply means for supplying (+) power and (-) power to the (+) electrode and (-) electrode, respectively; And a sample injection unit provided at one side of the center of the matrix; the sample injected into the sample injection unit includes centrifugation and electrophoresis at the same time as the flat circle rotates and DC power is supplied by DC power. It provides a sample separation device, characterized in that.

Description

Apparatus for separation of samples

Electrophoresis (also known as electrophoresis) refers to a phenomenon in which colloidal particles move toward one electrode when a DC voltage is applied by placing an electrode in a colloidal solution as is well known.

Electrophoresis is widely used to separate samples such as nucleic acids or proteins, and mainly injects a sample to be separated into a gel matrix and then applies a DC voltage to separate substances in the sample.

Gels used as matrices mainly use cross-linked polymers and determine their composition and porosity depending on the composition and characteristics of the material being analyzed. Proteins or relatively small nucleic acids (DNA) For RNA, oligonucleotides), a mixture of acrylamide and cross-linker of poly acrylamide forming a mesh network of various sizes is used as a matrix, and a relatively large In the case of nucleic acids, agarose is mainly used as a matrix. (Acrylate is different from polyacrylamide at this time, so care must be taken to prevent poisoning.)

When the sample is injected into the well inside the gel and a direct current voltage is applied, each component in the sample moves at different rates. The transfer ratio is mainly determined by the charge amount, size, and shape of each component. It is separated into components.

Various pretreatments may be performed for electrophoresis. Additives, such as sodium dodecyl sulfate (SDS) or sodium dodecyl phosphate (SDP), can be added to denature complex proteins to make them behave like long rods.

In some cases, the sample is first centrifuged, and the centrifuged sample is separated again through electrophoresis. Centrifugation is an analysis method using the principle that the components of the sample are arranged according to specific gravity when centrifugal force is applied to the sample. Generally, centrifugation is widely used to separate plasma and cellular components from whole blood or to separate cell components into red blood cells, white blood cells, and platelets. It is also used to isolate high-molecular-weight substances such as proteins, nucleic acids and lipids, including viruses.

In the case of such a centrifugation, a separate centrifuge is required, and there is a problem in that a part of the centrifuged sample needs to be electrophoresed again.

The present invention is to solve the above-mentioned problems, the problem to be solved by the present invention is to provide a sample separation device that can simultaneously perform centrifugation and electrophoresis using one device.

The present invention as a problem solving means described above,

A rotatable disk shaped platform;

A gel matrix formed on the upper portion of the platform in the same shape as that of the platform and configured for electrophoresis;

A (+) electrode provided at point A, which is any point of the matrix, and a (-) electrode provided at point B, which is point symmetrical with the point A with respect to the center of the matrix;

DC power supply means for supplying (+) power and (-) power to the (+) electrode and (-) electrode, respectively; And,

Sample injection unit provided on the central side of the matrix; including the sample injected into the sample injection unit can be carried out at the same time as the flat circle is rotated and the DC power is supplied by the DC power supply It provides a sample separation device characterized in that.

The DC power supply means,

A first disc provided concentrically with the platform and fixed to the platform regardless of rotation of the platform;

A second disk electrically separated from the first disk and provided in a concentric shape with the platform, and fixed to the platform regardless of rotation of the platform;

A first lead that rotates with the rotation of the platform while the first disc and the (+) electrode are connected to each other;

A second lead that rotates with the rotation of the platform while the second disc and the (-) electrode are connected to each other;

DC power supply section,

A first disc lead connecting the positive electrode of the DC power supply unit and the first disc, and a second disc lead connecting the negative electrode of the DC power supply unit and the second disc, wherein the first disc lead and the second disc lead wire are connected to each other. Is preferably disposed so as not to interfere with the rotating first and second conductors.

In the present invention, the DC power supply means,

A first disc provided concentrically with the platform and rotating together with the platform;

A second disc electrically separated from the first disc, the second disc being concentric with the platform, and rotating together with the platform;

DC power supply section,

A first disc lead connecting the positive electrode of the DC power supply unit and the first disc, and a second disc lead connecting the negative electrode of the DC power supply unit and the second disc, wherein the first disc lead and the second disc lead wire are connected to each other. May be disposed so as not to interfere with the rotating first and second conductors.

It is preferable to further include a centrifugal separator provided to extend in a direction away from the center of the matrix from the sample injection unit and to provide a space for centrifugation of the sample injected into the sample injection unit by the rotation of the platform.

The centrifugal separator,

More preferably, it is provided along an imaginary straight line passing through the center of the platform and the center of the sample injection unit.

According to the present invention having the above-mentioned problem solving means, by centrifugation and electrophoresis at the same time can be centrifugation and electrophoresis with a single equipment, the sample separation much more efficient than the case of centrifugation and electrophoresis separately It is possible to provide a sample separation device having the advantage of working.

In addition, since the electric force and the centrifugal force continue to act in the process of separating the sample can provide a higher resolution sample separation device than when centrifugation and electrophoresis separately.

Hereinafter, one embodiment of the present invention will be described with reference to the drawings to provide specific contents for the practice of the present invention.

1 is an exploded perspective view of a sample separation device according to an embodiment of the present invention, FIG. 2 is a combined perspective view of the sample separation device shown in FIG. 1, FIG. 3 is a cross-sectional view taken along the line III-III shown in FIG. It is sectional drawing of the IV-IV line shown in FIG.

First, each configuration of a sample separation device according to one preferred embodiment of the present invention will be described.

The sample separation device according to the present embodiment includes a platform 10, a matrix 20, a (+) electrode, a (−) electrode, a DC power supply unit 30, a sample injection unit 40, and a centrifuge unit 50. Is made of.

The platform 10 has a rotatable disk shape. In this embodiment, as shown in Fig. 1, there is an extension portion provided upwardly in the circumference portion, and has a shape similar to that of the whole.

The matrix 20 is formed in the same shape as the platform 10 on the platform 10, and is provided in a gel state for electrophoresis. The matrix 20 may selectively use agarose, acrylamide, or the like, depending on the type of sample to be separated. In case of isoelectric focusing, hydrogen ion concentration (pH) may be used. You can also make appropriate gradients.

The positive electrode is formed at a point A, which is a point in the matrix 20, the point A being indicated by a reference A in the drawing. The negative electrode is formed at a point B (indicated by reference numeral B) that is point symmetrical with the point A based on the center of the matrix 20 of the matrix 20. That is, the distance from the center of the matrix 20 to the positive electrode and the negative electrode are both equal to d.

The DC power supply means 30 includes a first disc 31, a first lead 32, a second disc 33 and a second lead 34, a first disc lead 35, and a second disc lead ( 36) and a DC power supply 37.

As shown in FIG. 1, the first disk 31 is provided concentrically with the platform, and is fixed so as not to rotate even when the platform 10 rotates. The through hole 311 provided at the center of the first disk 31 is a hole through which the output shaft 60 of the driving means (in this embodiment, the motor M) for rotating the platform 10 passes. Although not shown, a bearing may be provided between the output shaft 60 and the through hole 311. The first disc 31 has a groove 312 formed in the circumferential direction.

The first wire 32 is a wire that electrically connects the first disk 31 and the (+) electrode to each other and rotates together with the platform 10 when the platform 10 rotates. To this end, one end of the first conductive wire 32 rotates together with the platform 10 in contact with the groove 312 of the first disk 31.

Like the first disk 31, the second disk 33 is provided concentrically with the platform 10, and is fixed so as not to rotate even when the platform 10 rotates. The through hole 331 is also provided in the center of the second disc 33 and the output shaft 60 of the motor M passes through. Grooves 332 are also formed in the second disc 33 in the circumferential direction.

The first disk 31 and the second disk 33 are electrically separated from each other. In the drawing, the first disk 31 and the second disk 33 are shown to be in contact with each other. In this case, although not shown, an insulator is formed between the first disk 31 and the second disk 33. Have a plan.

The second conductive wire 34 is a conductive wire that electrically connects the second disc 33 and the negative electrode to each other and rotates together when the platform 10 rotates. To this end, as with the first conductive wire 32, one end thereof rotates together with the platform 10 in a state of touching the groove 332 of the second disk 33.

The DC power supply 37 may be a battery or the like as a DC power supply that is generally used.

The first disk lead 35 electrically connects the positive electrode of the DC power supply 37 and the first disk 31 so that DC power is supplied to the first disk 31.

The second disk lead 36 electrically connects the cathode of the DC power supply 37 and the second disk 33 so that DC power is supplied to the second disk 33.

The first disc lead 35 and the second disc lead 36 are arranged so as not to interfere with the rotating first lead 32 and the second lead 34 so as to achieve the above object. The second disc lead 36 is electrically connected to the second disc 33 through a through hole provided in the side surface of the first disc 31.

The sample injection part 40 is provided at one side of the center of the matrix 20 and is a groove formed in the matrix 20 as shown in the cross-sectional view of FIG. 4.

The centrifuge 50 is provided in a direction away from the sample injection unit 40 to the center of the matrix 20, the virtual passing through the center of the platform 20 and the center of the sample injection unit 40 It is provided along a straight line. The centrifuge 50 is a groove formed in the matrix 20, as shown in FIG.

The centrifuge 50 allows the sample to be centrifuged in the centrifuge 50 by filling a material having a density gradient.

Hereinafter, by explaining the mechanism of sample separation using the sample separation apparatus according to the present embodiment described, the function, operation and effects of each configuration will be described.

First, a sample to be separated is injected into the sample injection unit 40. When the sample is injected, by rotating the platform 10, the sample separated from the sample injection unit 40 is moved to the centrifuge 50, and centrifugation is performed. Due to the centrifugal force caused by the rotation of the platform 10, a relatively high specific gravity component is moved from a center of the platform 10 to a distant portion, and a relatively low specific gravity component remains near the center of the platform 10. The platform 10 is a bed for fixing the matrix 20 and interlocks with the motor M to rotate the matrix 20.

When the platform 10 rotates, electrophoresis is performed by supplying (+) power and (-) power to the (+) electrode and the (-) electrode by the DC power supply means 30, respectively. The sample centrifuged or centrifuged in the centrifuge 50 is moved through the matrix 20 by the electric force by the DC power supply means 30. At this time, since the direction of the electric force and the direction of the centrifugal force is different from each other, the separated sample is moved by the electric force and the movement by the centrifugal force at the same time, so that there is an advantage in that the separation of the finer sample by component.

In addition, in the case of taking such a configuration, the platform 10 may be rotated to stop the rotation of the platform 10 in the state where the centrifugal separation is performed first, and electrophoresis may be performed. In this case, both separation operations are performed in one device. There is a merit that can be easily implemented.

Hereinafter, another embodiment of the present invention shown in FIG. 5 will be described.

The sample separation device according to the present embodiment is the same as the sample separation device according to the previous embodiment.

The sample separation device according to the present embodiment includes a platform 110, a matrix 120, a (+) electrode (A), a (-) electrode (B), a DC power supply unit, a sample injection unit, and a centrifuge unit.

The platform 110, the matrix 120, the (+) electrode (A), the (-) electrode (B), the sample injection unit and the centrifugal separation unit are the same as the previous embodiment, so further detailed description will be omitted and sample injection Parts and centrifuges are omitted in the drawings.

DC power supply means of the present embodiment is the first disk 131, the first conductive wire 132, the second disk 133, the second conductive wire 134, the first disk conductor 135, the second disk conductor 136 ) And a direct current power source (137).

The first disk 131 and the second disk 133 is a disk provided concentrically with the platform 110, are separated from each other as electrical and rotated together by the platform 110 and the motor (M). .

The first conductor 132 and the second conductor 134 electrically connect the first disk 131 and the (+) electrode A, and the second disk 132 and the (-) electrode B, respectively. It rotates together with the first disk 131 and the second disk 133.

The first disk lead 135 and the second disk lead 136 electrically connect the positive electrode of the DC power source 137, the negative electrode of the first disk 131, the DC power source 137, and the second disk 133. Connect each.

The first disc lead 135 and the second disc lead 136 are disposed so as not to interfere with the rotating first lead 132 and the second lead 134. In this embodiment, As shown in FIG. 5, the first conductor 132 is disposed along the guide hole provided in the second disk 133, so that the first conductor 132 and the second conductor 134 rotate even when the first conductor 132 is rotated. It is comprised so that it may not interfere with the disk lead 135 and the 2nd disk lead 136. FIG.

In the above, an embodiment having a separate centrifuge has been described, but as shown in FIG. 6, when the platform is rotated even when the space indicated by the centrifuge is not provided separately, the sample injected into the sample injection unit is separated by the centrifugal force. It is moved by the centrifugal force inside, and centrifugal separation and electrophoresis may be performed by the centrifugal force. In this case, the matrix for electrophoresis is the centrifuge of the previous embodiment.

On the other hand, hepatic electrophoresis is an electrophoretic method of separating the nucleic acid of the polymer more efficiently by exchanging the voltage to a constant angle in the positive and negative directions with respect to the overall voltage axis. It may be embodied as a sample separation device to which the applied. Although not shown in the drawing, the (+) and (-) electrodes are further provided one by one along a straight line separated from the imaginary straight line connecting the (+) and (-) electrodes shown in FIG. It is also possible to further provide a DC power supply means for supplying a DC power to the additionally provided electrodes. As such, when the electrode and the DC power supply means are provided with one more set, and the DC power is alternately supplied by the two DC power supply means, so-called hepatocyte night electrophoresis (PFGE) type electrophoresis may be performed. It should be considered as being included in the technical idea of.

While the above provides specific details for the practice of the present invention by describing preferred embodiments of the present invention, the technical idea of the present invention is not limited to the described embodiments, and the scope does not deviate from the technical idea of the present invention. It can be embodied in various types of sample separation device inside.

1 is an exploded perspective view of a sample separation device according to an embodiment of the present invention.

Figure 2 is a perspective view of the combination of the sample separation device shown in FIG.

3 is a cross-sectional view taken along line III-III shown in FIG. 2;

4 is a cross-sectional view taken along the line IV-IV shown in FIG. 2;

5 is a cross-sectional view of a sample separation device according to another embodiment of the present invention.

Figure 6 is a simplified view showing a sample separation device according to another embodiment of the present invention.

** Explanation of symbols for the main parts of the drawing **

10: platform 20: matrix

30: DC power supply means 40: sample injection unit

50: centrifuge

Claims

A rotatable disk shaped platform;

Sample injection unit provided on the central side of the matrix; including the sample injected into the sample injection unit can be carried out at the same time as the flat circle is rotated and the DC power is supplied by the DC power supply Sample separation apparatus, characterized in that.

The method of claim 1,

The DC power supply means,

DC power supply section,

A first disc lead connecting the positive electrode of the DC power supply unit and the first disc, and a second disc lead connecting the negative electrode of the DC power supply unit and the second disc, wherein the first disc lead and the second disc lead wire are connected to each other. The sample separation device, characterized in that disposed so as not to interfere with the rotating first and second wire.

The method of claim 1,

The DC power supply means,

DC power supply section,

The method of claim 1,

Sample separation device further comprises a centrifugal separator provided to extend in a direction away from the center of the matrix from the sample injection unit and to provide a space for centrifugation of the sample injected into the sample injection unit by the rotation of the platform .

The method of claim 4, wherein

The centrifugal separator,

Sample separation device characterized in that provided along a virtual straight line passing through the center of the platform and the center of the sample injection unit.