KR20020064890A - Pin and ring type spotting device for sample solution - Google Patents

Abstract

Provided is a spotting apparatus for a sample solution by a pin and ring method that can spot valuable sample solution onto a gas without waste. In this spotting device 1, the lower part 15 of the vertical arm 4 adjacent the ring 5 and the lower part 15 of the vertical arm 4 when the pin 7 is in the lowered position. A space 28 is formed between the lower end portions 26 of the pin 7 opposite to. The space 28 has a length in the horizontal direction of 0.3 mm or more, or a length in the vertical direction of 5 mm or more.

Description

Pin and ring type spotting device for sample solution}

As a technique for creating a DNA chip by spotting a large number of DNA solutions on a gas surface such as a glass plate or a membrane filter, a spotting device 1 for a DNA solution by a pin and ring method shown in FIG. 4 is proposed. (Manufactured by GMS 417 Alain Genetic Microsystem).

As shown in FIG. 4, this spotting apparatus 1 has a solution holding mechanism 2 that holds a predetermined amount of DNA solution and a pin for spotting the solution held by the solution holding mechanism 2 on a gas. It consists of the mechanism 3. The former solution holding mechanism 2 includes a vertical arm 4 extending in the vertical direction, a ring 5 fixed to the lower end of the vertical arm 4, and a first lifting mechanism for raising the vertical arm 4 ( 6) consists of. The ring 5 has, for example, an inner diameter of about 1.2 mm. The latter pin mechanism 3 comprises a pin 7 thinner than the inner diameter of the ring 5, which is arranged along a vertical axis (center axis) passing through the center of the ring 5, and the pin 7 is a ring ( A second lifting mechanism 8 for moving the pin 7 between a raised position retracted upward of 5) and a lowered position where the lower end of the pin protrudes downward of the ring 5 through the ring 5; Has

In the case of spotting a DNA solution on a gas by using the spotting device 1, first, the vertical arm 4 is lowered by the first raising mechanism 6 so that the ring 5 is opened by the DNA solution 9. To soak (see steps (a) and (b) of FIG. 4). Thereafter, the vertical arm 4 is raised by the first raising mechanism 6 to lift the ring 5 from the DNA solution 9 (see step (c) in FIG. 4). In the state where the ring 5 is taken out of the DNA solution 9, a predetermined amount of the DNA solution 9 is held in the inner space of the thin ring 5 by the surface tension of the DNA solution 9. In the steps (a) to (c) thus far, the pins 7 and the ring 5 of the pin mechanism 3 are held in an elevated position, and the pins 7 are in contact with the DNA solution 9 without contact. It is maintained.

Next, the spotting apparatus 1 is moved above the base 10 (see step (d) of FIG. 4), and the pin 7 is lowered by the second lifting mechanism 8. As a result, the pin 7 penetrates through the DNA solution 9 held in the ring 5, and at this time, a small amount of the DNA solution 9 is almost retained at the lower end of the pin 7 (Fig. 4). See step (e)). When the pin 7 is further lowered by the second elevating mechanism 8 and the lower end of the pin 7 contacts the base 10, a small amount of DNA solution retained in the lower end of the pin 7 ( 9) is spotted on the base 10 (see step (f) of FIG. 4). When spotting is finished, the pin 7 is lifted by the second lifting mechanism 8, and returns to the lift position shown in Fig. 4 (d) again, waiting for the spotting of the same DNA solution 9 against another gas. do.

However, in the spotting apparatus 1 thus constructed, the DNA solution 9, which enters the gap between the pin 7 and the vertical arm 4, rises by a capillary phenomenon, and thus the inside of the ring 5 In some cases, the amount of the valuable DNA solution 9 remaining in the space was substantially reduced, so that the desired number of spots could not be performed.

In addition, there was a case where a large difference occurred between the spotting size initially spotted and the spotting size finally spotted due to the reduction of the DNA solution retained in the ring.

In addition, the DNA solution 9 remaining in the vertical arm 4 by the capillary phenomenon may be incorporated into other DNA solutions held in the ring 5 thereafter. Of course, since the vertical arm 4 and the like are usually cleaned when the required number of spots are finished, not all of the DNA solution raised along the vertical arm 4 is incorporated into other DNA solutions, but even when the cleaning is incomplete In such a case, it is considered that the remaining DNA solution will be incorporated into the DNA solution newly retained in the ring.

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a spotting apparatus for a sample solution by a pin and ring method for spotting a plurality of sample solutions, for example DNA solutions, on a substrate surface such as a glass plate.

Figure 1 (a) is a front view of the vertical arm and the ring used in the spotting apparatus of the sample solution by the pin and ring method according to the present invention.

Figure 1 (b) is a side view of the vertical arm and the ring used in the spotting apparatus of the sample solution by the pin and ring method according to the present invention.

Figure 1 (c) is a cross-sectional view of the vertical arm and ring used in the spotting apparatus of the sample solution by the pin and ring method according to the present invention.

1 (d) and 1 (e) are front views of a vertical arm and a ring for use in a spotting apparatus for a sample solution by a pin and ring method according to the present invention, respectively.

Figure 2 is a front view of the pin used for the spotting device for the sample solution by the pin and ring method according to the present invention.

3 (a) and 3 (b) are views showing the use states of the vertical arm and ring of Fig. 1 and the pins shown in Fig. 2;

4 (a) to 4 (g) are diagrams illustrating a spotting apparatus for a sample solution and a method of using the sample solution by a pin and ring method.

Here, an object of the present invention is to provide a spotting apparatus for a sample solution by a pin and ring method, in which a valuable sample solution held in a ring is not lost by capillary phenomenon.

Moreover, it aims at providing the spotting apparatus of the sample solution by a pin and ring system in which the other sample solution does not mix with the sample solution held by the ring.

In order to achieve this object, the spotting device of the sample solution by the pin and ring method according to the present invention is the bottom portion of the vertical arm adjacent to the ring, and the bottom of the vertical arm when the pin is in the lowered position The size of the vertical arm and the pin is determined so as to form a space between the lower end portions of the pin opposite to the portion having a horizontal length of 0.3 mm or more, or a vertical length of 5 mm or more.

In another form of this invention, the lower end part of the said vertical arm has a small cross section with a small horizontal cross section in the position adjacent to the said ring.

In another form of this invention, the said vertical arm and the said ring are formed by cutting the tube which has the same cross section as the said ring, The lower part of the said vertical arm which opposes the lower part of the said pin is formed in the inner surface of the said pipe. It has a plane parallel to the perpendicular plane to the tangent.

EMBODIMENT OF THE INVENTION Hereinafter, preferred embodiment of this invention is described with reference to attached drawing. Moreover, the spotting apparatus of the sample solution by the pin and ring system which concerns on this invention is characterized by the vertical arm 4 and the ring 5 of the solution holding mechanism 2, and the pin 7 of the pin mechanism 3. The basic configuration is the same as that of the spotting apparatus described above. Therefore, in the following description, only the structure of the vertical arm 4, the ring 5, and the pin 7 is described, and the whole structure of the spotting apparatus and its operation are not mentioned.

(1) vertical arm and ring

1 (a) to 1 (e) show an enlarged view of the vertical arm 4 of the spotting apparatus. As shown in these figures, the vertical arm 4 and the ring 5 are formed by cutting one straight straight stainless steel pipe 11. The machined vertical arm 4 is directed from one end (upper end) to the other end (lower end) with respect to the central axis 12 of the tube 11 (from top to bottom in Figs. 1 (a) and (b)), Most of the cross section of the pipe is cut off at the boundary between the upper end portion 13 having the cross-sectional shape of the pipe 11 before processing and the plane parallel to the central axis 12 of the pipe 11 as shown in Fig. 1 (d). The lower portion 15 of the cross section smaller than the intermediate portion 14 is cut off with the middle portion 14 and the majority of the tube cross section cut off at a boundary parallel to the central axis 12 of the tube as shown in FIG. Has

In this embodiment, as shown to FIG. 1 (d), the cross section of the part 14 'cut | disconnected in order to form the intermediate part 14 is half or more of a pipe cross section. In addition, in the cross section of the part 15 'cut | disconnected in order to form the lower part 15 as shown to FIG. 1 (e), the surface 16 of the side which opposes the central axis 12 is the tube 11 It is formed by a plane parallel to the surface in contact with the inner surface of the. However, this surface 16 may be a plane which contact | connects the inner surface of the pipe 11 mentioned above (in other words, the plane containing the tangent of an inner surface of a pipe).

The lower end part 15 is provided with the reduced cross section 17 which cut | disconnected the part adjacent to the ring 5 from both sides. This reduced cross section 17 is of various types (e.g., Fig. 1 (a) on one side) provided that the structural strength of the vertical arm 4 is no longer necessary in the reduced cross section 17. In the form of a cutout only on one side of the left side or the right side).

The ring 5 is sized in the direction of the central axis 12 so as to hold the required amount of sample solution in the inner space thereof.

(2) pin

2 shows an enlarged view of the pin 7. As shown, the pin 7 is formed by processing a rod made of stainless, and is directed from one end (upper end) to the other end (lower end) with respect to the central axis 20 of the pin 7 (from top to bottom in FIG. 2). ), Upper grip portion 21 (first portion), upper base portion 22 (second portion), taper portion 23 (third portion), lower base portion 24 (fourth portion), small diameter portion 25; 5th part), the micro diameter part 26 (6th part), and the minimum diameter part 27 (7th part, 27) are provided. Further, in the lower end surface of the minimum diameter portion 27, minute unevenness is formed by blasting using ceramic particles having a diameter of about 30 µm so that a predetermined amount of the sample solution can be reliably retained.

(3) state of use

The vertical arm 4 and the ring 5 configured as described above, and the pin 7 have a central axis 12 of the vertical arm 4 and the ring 5, as shown in FIGS. 3A and 3B. ) Is fixed to the first and second lifting mechanisms 6 and 8 (see FIG. 4) in a state where the central axis 20 of the pin 7 is matched with. As described above, the pin 7 (particularly, the micro diameter portion 26 and the smallest diameter portion 27) is penetrated through the DNA solution in the ring 5 in a state in which the sample solution is retained in the ring 5. Spot the sample solution retained on the bottom surface of (27) in the gas.

(4) Dimensions of vertical arm, ring and pin

The dimensions of the vertical arm 4 and the pin 7, in particular the size of the lower part 15 of the vertical arm 4 and the micro-diameter 26 of the pin 7, are shown in FIG. 3. And a space 28 of a predetermined size is formed between the lower portion 15 of the vertical arm 4, thereby preventing the sample solution from rising along the vertical arm in the space 28 by capillary action. It is decided.

Specifically, the experiment was performed by changing the size of the space 28 in various ways. As a result, when the space | interval (the magnitude | size of the horizontal direction of the space 28) of the ultra-small diameter part 26 and the vertical arm 4 which opposes this is 0.3 mm or more, the rise of the sample solution by a capillary phenomenon is extremely small. Although it was confirmed, the amount of increase was very small, and there was no problem in practical use. In addition, when the size of the horizontal direction of the space 28 is 0.3 mm, and the length (the size of the vertical direction of the space 28) of the microscopic diameter part 26 is 5 mm or more, the sample solution by a capillary phenomenon The increase of decreased further. As a most preferable combination, when the space | interval of the micro-diameter part 26 and the vertical arm 4 is 0.45 mm or more and the length of the vertical direction of the micro-diameter part 26 is 7.5 mm or more, the raise of the sample solution by a capillary phenomenon Was almost completely restrained. It was also confirmed that the reduced cross section 17 formed on the lower end 15 of the vertical arm 4 is effective in suppressing the rise of the sample solution due to the capillary phenomenon. In view of the above, the distance between the micro-diameter portion 26 and the vertical arm 4 is 0.3 mm or more, preferably 0.45 mm or more, and the length in the vertical direction of the micro-diameter portion 26 is 5 mm or more, preferably 7.5 mm or more. I can understand.

Regarding preferred embodiments of the vertical arm 4, the ring 5, and the pin 7, the dimensions of their respective portions are shown below.

① vertical arm and ring (see Fig. 1)

D1: 1.8 mm

D2: 1.2 mm

L1: 5.5 mm

L2: 6.0 mm

L3: 5.0 mm

L4: 1.0 mm

W2: 0.6 mm

W3: 0.3 mm

② pin (see Figure 2)

L1: 4.5 mm

L2: 10.0 mm

L3: 1.0 mm

L4: 6.0 mm

L5: 12.5 mm

L6: 7.5 mm

L7: 1.0 mm

ø1: 2.3 mm

ø2: 4.0 mm

ø3: 2.3 mm

ø4: 0.6 mm

ø5: 0.3 mm

ø6: 0.20, 0.12, 0.08 mm (changes depending on spot diameter)

(5) gas

The gas to which the sample solution is spotted using the spotting apparatus of the present invention is not particularly limited as long as it is of a substantially insoluble material. Examples include paper, nylon, cellulose, cellulose derivatives such as nitrocellulose, plastics and plastic derivatives such as polycarbonate, polystyrene and polypropylene, magnetic or nonmagnetic metals, quartz and glass, and the like. In addition, the shape is not specifically limited in the range which the spotting of the sample solution by the said apparatus is possible, The plate-shaped or the gas on a film | membrane which has a homogeneous thickness is preferable in using the apparatus of this invention. .

(6) sample solution

The sample solution spotted using the spotting apparatus of the present invention is not particularly limited, but any solution containing any sample that is desired to be spotted on a gas may be used. The apparatus of the present invention is particularly suitable for the immobilization operation of biologically related materials in fields such as biochemistry and molecular biology. Sample solutions used in the operation include, for example, solutions containing biologically relevant materials such as nucleic acids, proteins, peptides, sugars, drugs. Gases onto which such biorelevant materials have been immobilized are useful for the search and detection of substances having affinity for the materials.

(7) other

In the above description, the vertical arms, rings, and pins were formed by processing stainless steel pipes and rods, respectively, but the materials constituting them are not limited to stainless, but non-ferrous metals including plastics and ceramics. You may form.

In addition, these are not limited to the thing processed by the integrally formed pipe | tube and rod, but may be formed by assembling separately created parts.

As is clear from the above description, according to the spotting apparatus of the sample solution by the pin and ring system according to the present invention, the precious sample solution held in the ring can be spotted on the gas without waste. In addition, the spotting size of the sample solution spotted on the gas becomes almost constant size, shape. In addition, since the sample solution does not rise and remain in the vertical arm due to the capillary phenomenon, no other DNA solution is mixed into the sample solution held in the ring.

Claims (5)

A vertical arm 4 extending in the vertical direction, A sample solution holding ring 5 disposed along a horizontal plane and fixed to a lower end of the vertical arm 4, The lower end portion penetrating the ring 5 and the raised position in which the lower end is positioned parallel to the vertical arm 4 along the central axis 12 of the ring 5 and on the side of the vertical arm 4. Has a rod-like pin 7 which moves between the lowered positions located below the ring 5, The sample solution held in the space inside the ring 5 by holding the sample of the sample solution 9 in the space inside the ring 5 by the surface tension of the sample solution 9 ( The sample of the sample solution 9 held in the lower end of the pin 7 by holding the sample of the sample solution 9 at the lower end of the pin 7 by passing the pin 7 through the sample of 9). In the spotting apparatus 1 of the sample solution by the pin and ring system which attaches to the surface of the base 10, The pin opposite the lower portion 15 of the vertical arm 4 and the lower portion 15 of the vertical arm 4 when the pin 7 is in the lowered position. Between the lower end part 26 of (7), the size of the said vertical arm 4 and the pin 7 is defined so that the space length of a horizontal direction may be 0.3 mm or more. And a spotting device for a sample solution by a ring method. A vertical arm 4 extending in the vertical direction, A sample solution holding ring 5 disposed along a horizontal plane and fixed to a lower end of the vertical arm 4, The lower end portion penetrating the ring 5 and the raised position in which the lower end is positioned parallel to the vertical arm 4 along the central axis 12 of the ring 5 and on the side of the vertical arm 4. Has a rod-like pin 7 which moves between the lowered positions located below the ring 5, The sample solution held in the space inside the ring 5 based on the surface tension of the sample solution 9 and held in the space inside the ring 5. The pin 7 is penetrated through the sample of (9) to hold the sample of the sample solution 9 at the lower end of the pin 7, and the sample solution 9 held at the lower end of the pin 7 In the spotting apparatus 1 of the sample solution by a pin and ring system which attaches a sample to the surface of the base 10, The pin opposite the lower portion 15 of the vertical arm 4 and the lower portion 15 of the vertical arm 4 when the pin 7 is in the lowered position. Between the lower end portion 26 of (7), the vertical arm 4 and the pin 7 are sized so as to form a space 28 in the vertical direction of 5 mm or more, and Spotting device for sample solution by the ring method. Pin and ring system according to claim 1 or 2, characterized in that the lower end portion of the vertical arm (4) has a small cross-sectional area (17) having a small cross-sectional area at a position adjacent to the ring (5). Spotting apparatus of the sample solution by. 4. The vertical arm (4) and the ring (5) according to any one of claims 1 to 3 are formed by cutting a pipe (11) having the same cross section as the ring (5). The lower portion 15 of the vertical arm 4 opposite the lower portion 26 of (7) has a plane 16 parallel to the vertical surface in contact with the inner surface of the tube 11. Spotting device for sample solution by pin and ring method. The spotting apparatus of a sample solution according to any one of claims 1 to 4, wherein the sample is DNA.