JP3898526B2 - Spotting nozzle - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a spotting nozzle that receives from a holding tank that holds a transferred liquid and transfers a small amount of transferred liquid droplets onto a substrate such as a biochip.
[0002]
[Prior art]
In recent years, biochips have begun to be used for disease risk diagnosis, rapid and accurate disease diagnosis, tailor-made medicine, and the like. Biochips need to be accurately spotted and arranged from hundreds to tens of thousands of reagents such as DNA fragments on a few cm square glass or silicon plate (substrate). It is commercially available.
[0003]
Conventionally, there are the following two methods for spotting picoliter droplets on a plate.
1. A method in which the stamping nozzle is directly applied to the plate and the liquid adhering to the nozzle tip is continuously transferred onto the plate.
2. A method in which the liquid filled in the closed area is pressurized and sprayed in a small amount, and continuously spotted on the plate. (For example, pump pressurization method, piezoelectric element pressurization method, electric field acceleration method, etc.)
There is.
[0004]
[Problems to be solved by the invention]
However, the conventional technique has the following problems.
1. In the case of a stamping nozzle, the tip of the nozzle is worn because the nozzle and the plate are in contact with each other. Further, there is a possibility that the plate may be broken or scratched, and there is a risk that contamination will occur from them. Further, when spotting is performed in a place where a sample is placed in advance or by stacking different liquids, the sample on the plate may adhere to the nozzle, and in some cases, the sample may be damaged.
[0005]
2. In the case of closed pressure injection, a closed nozzle can spot droplets continuously, but cannot quickly spot different liquids at designated locations. Since the inside of the nozzle is a closed system, it is difficult to clean and inject the nozzle when a different liquid is used next time. In addition, even if the number of spots required is small, it is necessary to fill a certain amount of liquid in the closed container, and there is a lot of waste of liquid to be used. Poor versatility.
[0006]
For these reasons, it is difficult to clean the nozzles and inject liquid when spotting different liquids, or the problem of nozzle tip wear and plate damage. In addition, there is a problem that switching between different liquids is inconvenient.
[0007]
In view of these problems, an object of the present invention is to provide a spotting nozzle that can freely select different liquids and perform spotless picoliter order spotting.
[0008]
[Means for Solving the Problems]
A spotting nozzle according to the invention described in claim 1 is a spotting nozzle that receives a transfer liquid from a liquid holding tank and transfers a small amount of a liquid drop of the transfer liquid onto a substrate.
An open hole communicating the inside of the nozzle hole from the side at a position spaced a predetermined distance from the tip of the nozzle hole formed at the tip of the spotting nozzle, and
A tip thin tube portion that forms a nozzle hole portion closer to the nozzle hole tip than the position;
High-pressure air supply means for supplying high-pressure air from the nozzle hole portion behind the position toward the tip thin tube portion;
When the nozzle tip is immersed in a holding tank, the liquid to be transferred is held in the tip capillary, and the droplet is ejected onto the substrate by the high-pressure air of the high-pressure air supply means and transferred. is there.
[0009]
The spotting nozzle according to the invention described in claim 2 further includes low-pressure air supply means for generating a steady flow of air from the rear nozzle hole portion according to claim 1 toward the opening hole,
When the nozzle tip is immersed in a holding tank, liquid is prevented from entering from the open hole to the rear nozzle hole.
[0010]
A spotting nozzle according to a third aspect of the present invention is characterized by comprising air generating means shared by the high-pressure air supply means and the low-pressure air supply means according to the first or second aspect. .
[0011]
A spotting nozzle according to a fourth aspect of the invention is characterized in that the high-pressure air supply means and the low-pressure air supply means according to any one of the first to third aspects each have a pressure adjusting means. It is.
[0012]
A spotting nozzle according to a fifth aspect of the present invention is a spotting nozzle according to any one of the first to fourth aspects, wherein the cleaning liquid is discharged from the opening hole and the distal end pipe portion by air pressure as an operation during the cleaning of the nozzle front end portion. A suction means for sucking into the nozzle is provided.
[0013]
According to a sixth aspect of the present invention, there is provided a spotting nozzle that cleans the tip of the nozzle with at least one of the high-pressure air supply means and the low-pressure air supply means according to any one of the first to fifth aspects. Sometimes used as dry air.
[0014]
DETAILED DESCRIPTION OF THE INVENTION
In the present invention, the opening hole that communicates the inside of the nozzle hole from the side at a position spaced a predetermined distance from the tip of the nozzle hole formed at the tip of the spotting nozzle, and the nozzle more than the above position And a high-pressure air supply means for supplying high-pressure air from the nozzle hole portion rearward of the position toward the front-end thin tube portion. When immersed in the holding tank, the transfer target liquid is held in the tip thin tube portion, and droplets are ejected onto the substrate by the high pressure air of the high pressure air supply means and transferred. As a result, the spotting nozzle of the present invention is capable of freely selecting different liquids and enabling spotless picoliter order spots without contact.
[0015]
FIG. 1 is an explanatory view showing the configuration of a spotting nozzle according to an embodiment of the present invention. As shown in the figure, the spotting nozzle (10) according to the present invention is located laterally at a position separated by a predetermined distance L from the tip of a nozzle hole (12) consisting of a narrow tube having a diameter d that passes through the substantial center of the spotting nozzle. An open hole (13) communicating with the nozzle hole, and a tip thin tube section (11) that forms a nozzle hole portion closer to the nozzle hole tip side than this position.
[0016]
That is, in consideration of the amount of spot required from the nozzle tip, the tip capillary (11) (liquid holding area) that secures the diameter d and length L is opened to the outside from the side at that position. An open hole (13) is provided. The arrangement of the open holes (13) can be arbitrarily selected in consideration of cleaning and the like. For example, one open hole may be provided on the side, and a plurality of open holes may be provided, for example, radially.
[0017]
Injecting the retentate retained in the tip capillary as droplets is performed by high pressure air supply means for supplying high pressure air from the rear nozzle hole toward the tip capillary. For example, as shown in FIG. A, a high pressure air supply means (18a) comprising an air generation means (14a) and a high pressure air adjustment means (16a) that hardly reduces the high pressure of the air generated by the air generation means (14a). ) To supply high-pressure air from the rear of the nozzle hole (12) in the direction of the tip thin tube portion. By supplying air having a pressure higher than that passing through the open hole (13) provided on the side, the retentate can be transferred onto the substrate as droplets in a non-contact state.
[0018]
In the present invention, preferably, the same air generating means (14a) also causes a steady flow of air from the rear nozzle hole (12) portion toward the open hole (13) (and the tip narrow tube portion (11)). ing. That is, when the high pressure air from the air generating means (14a) passes through the low pressure air adjusting means (17a) as the low pressure supply means (19a) comprising the air generating means (14a) and the low pressure air adjusting means (17a). In addition, the low-pressure steady air always flows from the rear nozzle hole (12) portion to the open hole (13). Thereby, when the nozzle tip is immersed in the holding tank, the liquid can be prevented from entering the rear nozzle hole (12).
[0019]
That is, the air generating means (14a) supplies air from the rear of the nozzle hole (12), but in particular, when the spot liquid is filled, the wetting at the rear liquid surface of the tip thin tube section (11), which is the liquid holding area. Alternatively, low-pressure air of a level that prevents the liquid level from rising to the wall surface due to capillarity is constantly supplied by the low-pressure air adjusting means (17a), and the air in the liquid holding area is spotted on the plate. The pressure is supplied by the air generating means (14a), and the high pressure is adjusted by the air adjusting means (17a), so that a spot in the picoliter order can be achieved without contact.
[0020]
The air generating means (14a) for generating high-pressure air toward the tip thin tube portion (11) is air directed from the rear nozzle hole (12) portion toward the open hole (13) (and the tip thin tube portion (11)). However, each of the two air generators (ie, the high pressure air generating means and the low pressure air generating means) generates air of a desired pressure and finely adjusts the air by the air adjusting means. Thus, an air flow rate and an air pressure that match the conditions of the high pressure air and the low pressure steady flow may be supplied.
[0021]
FIG. b is an explanatory view showing the configuration of another air supply means introduced into the spotting nozzle. As shown in the figure, when jetting the retentate retained in the tip capillary section as droplets, high-pressure air generating means (14) is provided to supply high-pressure air from the rear of the nozzle hole (12) toward the tip capillary section. ) And high-pressure air adjustment means (16), and the retentate is supplied by supplying high-pressure air that passes through the open hole (13) provided on the side. The droplet can be transferred onto the substrate in a non-contact state.
[0022]
In the present invention, it is preferable to further include a low-pressure air supply means (19) for generating a steady flow of air from the rear nozzle hole (12) portion toward the open hole (13) (and the tip narrow tube section (11)). That is, a low pressure air supply means (19) comprising a low pressure air generation means (15) and a low pressure air adjustment means (17) is provided. Thereby, when the nozzle tip is immersed in the holding tank, the liquid can be prevented from entering the rear nozzle hole (12).
[0023]
In other words, air is supplied from the rear of the nozzle hole (12), but when filling the spot liquid, the liquid surface area on the liquid surface due to wetting or capillary action at the rear liquid surface of the tip capillary (11), which is the liquid holding area, is provided. Low-pressure air that prevents rising is constantly supplied from the low-pressure air supply means (19), and on the plate, the high-pressure air supply means (18) supplies air pressure that allows droplets in the liquid holding area to be spotted. Supplying it enables spotless picoliter order spots.
[0024]
By the way, the amount of liquid to be held is determined by the diameter d and the length L of the tip thin tube portion (11) as the liquid holding area of the tip portion. Assuming that the liquid volume determined by the diameter d and the length L is the rated liquid volume, the flow rate of the steady-state low-pressure air, the pressure waveform of the high-pressure air, etc. are further controlled with respect to the rated liquid volume. Fine adjustment of spotted droplets is possible.
[0025]
For example, when the liquid is held, the pressure of the steady air flow is set to be large by the low-pressure air adjusting means (16), so that the position of the rear liquid surface portion is lowered in the tip direction, so that it is held slightly less than the rated liquid amount. be able to. When spotting, high-pressure air is supplied by the high-pressure air supply means. By adjusting the supply pressure and supply amount with the high-pressure air adjustment means (17), an amount smaller than the rated liquid amount can be spotted. Can do. For example, by spraying high pressure air at high and short impulses (that is, by reducing the supply amount), for example, 1/2, 1/3, 1/4, etc. of the rated liquid amount is spotted. Is possible.
[0026]
In the nozzle of the present invention, since the tip end portion is opened by the open hole (13), the tip thin tube portion holding the transferred liquid can be easily cleaned with running water or ultrasonic waves. More preferably, by further providing a suction means (20) for sucking a small amount of cleaning water into the nozzle hole (12) on the rear side from the open hole at the time of cleaning, by extruding the sucked cleaning water with high-pressure air. Also, there is an advantage that the inside of the distal thin tube portion (11) can be more easily cleaned.
[0027]
In addition, there is an advantage that the cleaned nozzle tip portion can be easily dried by using the supply air of the air supply means as dry air when cleaning the nozzle tip portion.
[0028]
【Example】
FIG. 2 is an explanatory diagram showing a step of spotting droplets by the nozzle shown in FIG. As shown in step a, low pressure air is supplied into the nozzle (10). As shown in step b, the tip of the nozzle (10) is immersed in a liquid holding tank (21) holding a transfer target liquid (22) in a state where low-pressure air is supplied. At this time, the liquid to be transferred (22) is filled in the tip capillary (11) due to wetting or capillary action, but the nozzle hole (12) behind the open hole (13) for air release is caused by low pressure air. Liquid cannot enter.
[0029]
As shown in step c, when the tip of the nozzle (10) is pulled up from the liquid to be transferred (22), the liquid to be transferred (22) is held in the tip capillary portion (11) by wetting or capillary action. Since the low-pressure air is discharged from the opening hole (13), the transfer target liquid (22) does not escape from the tip thin tube portion (11). In addition, since the air is in a steady flow, a constant liquid amount is always held in the tip thin tube portion (11).
[0030]
As shown in step d, the nozzle (10) is moved to the spot area. High-pressure air is supplied at a predetermined position, and the held liquid to be transferred (22) is dropped onto the substrate (23) directly below.
[0031]
FIG. 3 is an explanatory view showing a cleaning process of the nozzle shown in FIG. As shown in step e, the nozzle tip is immersed in the cleaning liquid (32) in the cleaning tank (31) while supplying low-pressure air. At this time, the cleaning liquid (32) is injected into the tip capillary (11) below the opening hole (13) for air release to be cleaned. The suction means (20) sucks the air pressure in the nozzle hole (12) behind the open hole (13), so that a small amount of washing water can be removed from the nozzle hole (13) behind the open hole (13). It may be sucked into 12) and extruded with high-pressure air in the direction of the tip thin tube. This makes it easier to clean the inside of the distal thin tube section (11).
[0032]
As shown in step f, the nozzle (10) is lifted from the cleaning tank (31), and high pressure air is supplied to dry the inside of the nozzle.
[0033]
As described above, the steady flow of low-pressure air can prevent wetting of the rear liquid surface portion of the tip capillary or the rise of the liquid level due to capillary action, and a constant liquid can be always maintained. As a result, a constant liquid can always be held and spotted on the tip capillary (11). It is also possible to increase the size of the steady flow of the low-pressure air so as to hold a smaller amount of liquid than the rated liquid amount in the tip thin tube portion.
[0034]
Further, since the nozzle tip is open, cleaning and drying can be performed easily. Therefore, a different liquid can be spotted at an arbitrary place every time. Since wasteful liquid is not consumed, less valuable liquid is required and it is economical. For example, although reagents used in DNA and protein tests are expensive, they are economical because they have spots in the order of picoliters.
[0035]
Furthermore, the amount of spot can be controlled by adjusting the pressure waveform of high-pressure air when spotting. That is, by adjusting the pressure of the high-pressure air to a high and short impulse shape so as not to dripping all of the liquid held in the tip thin tube portion, this is adjusted and, for example, 1/2 of the rated liquid amount is adjusted. , 1/3, 1/4, etc. can be spotted.
[0036]
In addition, since the nozzle can be easily washed and dried, different liquids can be spotted quantitatively at any place without contact. As a result, variations in biochip production are increased, enabling rapid and precise inspection. There is no wear or impact because the nozzle and plate do not contact.
[0037]
【The invention's effect】
As described above, the present invention has an effect that a different liquid can be freely selected and a spot of picoliter order can be performed without contact.
[Brief description of the drawings]
FIG. 1 is an explanatory diagram showing a configuration of a spotting nozzle according to an embodiment of the present invention.
FIG. 2 is an explanatory diagram showing a step of spotting droplets by the nozzle shown in FIG. 1;
FIG. 3 is an explanatory view showing a cleaning process of the nozzle shown in FIG. 1;
[Explanation of symbols]
(10)… Spotting nozzle,
(11)… the tip tubule,
(12)… Nozzle hole,
(13)… Open hole,
(14a)… Air generating means
(16a) ... high pressure air adjusting means,
(17a) ... Low pressure air adjusting means,
(18a)… high pressure air supply means,
(19a)… low pressure air supply means,
(14) ... High pressure air generating means
(15) ... Low pressure air generating means,
(16) ... High pressure air adjusting means,
(17) ... Low pressure air adjustment means,
(18) ... High pressure air supply means,
(19) ... Low pressure air supply means,
(20) ... suction means,
(21)… Liquid holding tank,
(22)… Transfer solution,
(23)… Substrate,
(31)… Cleaning tank,
(32)… Cleaning liquid,

Claims (6)

In a spotting nozzle that receives a transferred liquid from a liquid holding tank and transfers a small amount of transferred liquid droplets onto a substrate,
An open hole communicating the inside of the nozzle hole from the side at a position spaced a predetermined distance from the tip of the nozzle hole formed at the tip of the spotting nozzle, and
A tip thin tube portion that forms a nozzle hole portion closer to the nozzle hole tip than the position;
High-pressure air supply means for supplying high-pressure air from the nozzle hole portion behind the position toward the tip thin tube portion;
A spotting nozzle that retains a liquid to be transferred in a tip thin tube portion when the nozzle tip is immersed in a holding tank, and jets and transfers droplets onto the substrate with high-pressure air from the high-pressure air supply means. . Low pressure air supply means for generating a steady flow of air from the rear nozzle hole portion toward the opening hole,
The spotting nozzle according to claim 1, wherein when the nozzle tip is immersed in a holding tank, liquid is prevented from entering the rear nozzle hole. The spotting nozzle according to claim 1 or 2, further comprising air generating means shared by the high-pressure air supply means and the low-pressure air supply means. The spotting nozzle according to any one of claims 1 to 3, wherein each of the high-pressure air supply means and the low-pressure air supply means includes a pressure adjusting means. 5. The apparatus according to claim 1, further comprising suction means for sucking a cleaning liquid into the nozzle from the open hole and the tip tube portion by air pressure as one operation at the time of cleaning the nozzle tip portion. The spotting nozzle described. 6. The spotting nozzle according to claim 1, wherein at least one of the high-pressure air supply means and the low-pressure air supply means is used as dry air when cleaning the nozzle tip.

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