JPS6311863A - Method for depositing liquid specimen in spot form - Google Patents

Abstract

PURPOSE:To accurately control suction/emitting quantity, by a method wherein the leading end of a spot-deposition chip is preliminarily immersed in a liquid specimen when spot-deposition is performed by the same spot-deposition chip so as to leave an interval and the specimen in the spot-deposition chip is emitted at the time of the next spot-deposition before the specimen is again sucked. CONSTITUTION:When spot-deposition is performed by the same spot-deposition chip 53 so as to leave an interval, a spot-deposition arm 55 is revolved by a chip moving means 80 after first spot-deposition is completed to allow the chip 53 to move to a sucking position and the leading end of the chip 53 is immersed in the liquid specimen in a specimen container 52 by a chip up-and- down control means 60 to be prevented from drying. Next, when second spot- deposition is performed, the chip 53 is raised above the liquid specimen by th means 60 and the specimen in the space 53a of the chip 53 is perfectly emitted by a suction/emitting means 90. Thereafter, the chip 53 is again moved downwardly to be immersed in the liquid specimen and a predetermined quantity of the liquid specimen is sucked in the space 53a by the means 90. By this method, even when the liquid specimen is removed along with the emitted liquid specimen and clogging can be prevented effectively.

Description

[Detailed Description of the Invention] (Industrial Application Field) The present invention relates to a method for dotting a fixed amount of a sample liquid onto a chemical analysis slide (hereinafter also simply referred to as a slide) having a predetermined reagent layer. It is.

(Prior Art) Qualitative or routine analysis of specific chemical components in a liquid sample is a commonly performed operation in various industrial fields. In particular, quantitative analysis of chemical components or formed components in biological body fluids such as blood and urine is extremely important in the biochemical and clinical fields.

In recent years, dry type slides have been developed that allow quantitative analysis of specific chemical components or 1-forming components contained in a sample solution simply by applying small droplets of the sample solution. Publication No. 53-21677, Japanese Patent Application Publication No. 55-1643
No. 56, etc.) has been put into practical use. Using these slides allows sample liquid analysis to be performed more easily and quickly than conventional wet analysis methods, so their use is particularly important in medical institutions, laboratories, etc. that need to analyze a large number of samples. This is preferable.

To analyze chemical components in a sample solution using such a slide, the sample solution is deposited on the slide and then kept at a constant temperature (incubation) for a predetermined period of time in an incubator. The slide undergoes a color reaction (pigment formation reaction), and then the slide is irradiated with measurement light containing a wavelength pre-selected based on the combination of sample components and reagents contained in the reagent layer of the slide to determine its reflected optical density. This is used to quantitatively analyze the chemical components mentioned above.

When performing such an analysis, a predetermined amount of sample liquid must be spotted onto the reagent layer of the slide at an accurate Klm. This is because if the amount of the sample liquid differs from the predetermined amount, the reflected optical density will differ and the analysis accuracy will also decrease. For this reason, various pipettes and the like have been devised so that when spotting and supplying a sample liquid, it is possible to spot a predetermined amount ω correctly.

For example, in a bed like this, a spotting tip is attached to the tip of the bevet 1, a predetermined amount of sample liquid is sucked into the spotting tip, and then a predetermined amount of sample r[liquid is poured into the reagent layer of the slide. There is a droplet on the top that is used for dotting. Many of these pipettes 33 use a piston-cylinder i structure to aspirate a predetermined amount of sample liquid into the dispensing tip and discharge it. To aspirate the sample liquid into the spotting tip and discharge it to the reagent layer using such a pipette, first insert the tip of the spotting tip into the sample liquid, and then use the piston-cylinder mechanism etc. A predetermined amount of sample liquid is sucked and held in the spotting tip, and then the tip of the spotting tip is positioned on the reagent layer of the slide, and the sample liquid in the spotting tip is dispensed onto the reagent layer using a piston-cylinder mechanism, etc. .

(Problem to be Solved by the Invention) When the sample solution in the spotting tip is dispensed onto the reagent layer as described above, the spotting tip is opened for a predetermined time or more while the sample solution is sucked into the spotting tip. If left in the air, the sample solution dries and the tip of the spotting tip becomes clogged, making it impossible to spot. In particular, the amount of liquid to be spotted onto the reagent layer of a chemical analysis slide is small, and the opening at the tip of the spotting tip for aspirating and discharging this liquid is small and prone to clogging.

In this case, in the case of a disposable tip that only performs one spotting with one spotting tip, it is recommended to do the spotting immediately after aspirating the sample liquid to shorten the time that the tip of the tip is exposed to the air. Although it is easy, in the case of continuous spotting, in which multiple doses of sample solution are aspirated into one spotting tip and the sample solution is divided into multiple doses and spotted onto the reagent layer of multiple chemical analysis slides,
The time interval between the previous dot and the next dot tends to be large,
The above clogging problem is likely to occur.

(Means for Solving the Problems) The present invention solves the above problem, and is aimed at preventing clogging due to drying of the sample liquid at the tip of the spotting tip when spotting is performed multiple times using the same spotting tip. The purpose is to provide a method of spotting that does not cause the occurrence of such problems, and the following method is used as a means for this purpose.

In this method, when spotting is performed multiple times and the time interval from the previous spotting to the next spotting is longer than a certain value, the tip of the spotting tip is inserted into the sample container after the previous spotting is completed. For the next spotting, the tip is pulled out of the sample liquid and the sample liquid left in the spotter tip is discharged into the sample container, and then the tip is immersed in the sample liquid again. A predetermined amount of sample liquid is sucked and held from the tip into the spotting tip, and then the sample liquid in the spotting tip is spotted onto the reagent layer of the chemical analysis slide. .

(Function) When the above spotting method is used, when the spotting interval is long, the tip of the spotting dip is immersed in the sample solution after the previous spotting is completed and the sample is collected at the tip. This prevents clogging due to liquid drying, and for the next spotting, by discharging the sample liquid in the spotting tip and then sucking the sample liquid into the spotting tip again, any remaining dried sample can be removed from the tip. Remove this if there is one. In addition, even if the air inside the suction/discharge mechanism expands or contracts due to temperature changes due to the increased spotting interval, these problems can be eliminated by not re-suctioning the sample liquid after discharging the sample liquid. influence can be eliminated.

(Example) Hereinafter, a specific method of the present invention will be described with reference to the drawings.

FIG. 1 is a perspective view showing an example of a chemical analysis apparatus having an automatic spotting device 50 used for carrying out the spotting method according to the present invention.
First, this 5A question will be explained, and then the spotting method of the present invention using this device will be specifically explained.

This device includes a cartridge 11, an incubator 201, a conveyance/insertion means 40, and an automatic spotting device 50 on a main body 10.
The top view of FIG. 2 shows these as seen from above with the cover plate 16 covering the main body 10 removed. Additionally, as shown in Figure 1, this device includes:
A display unit 14 that displays measurement data during measurement. An operation key 15d3 for operating this display etc. and a magnetic disk insertion section 13 for recording are installed.
In FIG. 2, these are omitted.

The cars 1 to 11 store a plurality of unused chemical analysis slides 1 stacked on top of each other, and these chemical analysis slides 1 are pushed out one by one from the lowest one to the next 1'i (
It is extruded in the direction of the arrow in FIG.

On the right side of this cartridge 11 is an incubator 20.
are located in the incubator 20, and in the incubator 20 there are a plurality of storage chambers 21, 21, . 21 is formed. In front of the incubator 20, a tray 29 is arranged to receive used slides discharged from the storage chamber 21. Further, an incubator 20 is provided below the incubator 20.
A reading head (not shown) for optical density measurement is disposed on the lower surface of the optical density sensor, and is slidable laterally (in the direction of arrow C). Note that this sliding is performed by being driven by, for example, a linear motor or the like on a rail disposed laterally below the incubator 20, and this rail extends to the bottom surface of the cartridge 11, so that the reading head is placed on the bottom of the cartridge 11.
It can be slid to a position facing the lower surface of the slide 1, and can face the chemical analysis slide 1 at the bottom of the cartridge. With this, it is possible to not only measure the reflected optical density of the slides 1 stored in each storage chamber 21 of the incubator 20 using the reading head, but also perform fog photometry to measure the reflected optical density of unused slides 1. is now possible.

This incubator 10 has a built-in heater (not shown), and is capable of maintaining (incubating) the chemical analysis slides 1 held in the storage chambers 21, 21, 21 at a constant temperature. The chemical analysis slide 1 consists of a dry multilayer film in which a support, a reagent layer, and a developing layer are laminated in this order in a frame with a circular hole for dropping a liquid sample, and urine, blood, etc. are placed on this film. A sample is dropped at a predetermined temperature and kept at a constant temperature in an incubator 10 to cause a color reaction.

On the other hand, behind the ink-L beta 20, a conveyance/insertion means 40 that faces the entrance opening 21a of the storage ¥21 and is slidable in the lateral direction (in the direction of arrow C) is disposed. In this F first, the conveyance/insertion means 4 is placed on a rail 49 extending laterally.
0 is driven by, for example, a linear motor, etc., and is used not only for the incubator 20 but also for the cartridge 1.
1 (the position indicated by the chain line X in FIG. 2).

For this reason, the chemical analysis slide 1 pushed out from the cartridge 11 by the pushing lever 12 is received by the transport/insertion means 40 slid to the position indicated by the chain line X, and is transported to a predetermined position or thereafter. This slide 1 can be inserted into a predetermined storage chamber 21.

At the rear of the conveyance/insertion means 40, an automatic spotting device 50 having a rotatable spotting arm 55 is disposed as shown by arrow B at J3 in FIG. This spotting arm 55 has a removably attached spotting tip 53 at its tip, and this tip rotates in the direction of arrow B to a suction position located above the sample container 52 placed on the main body 10. (the position shown by the chain line in the figure) and the 16-point landing position (the position shown by the solid line in the figure) located above the spotting hole 19 formed in the cover plate 16 on the main body 10. ing. Spotting hole 19
The receiver is removed from the cartridge 11 by the transport/insertion means 40 and transported on the rail 49, and then placed at a predetermined position (the position shown by the solid line Y in FIG. 2C) for spotting the sample liquid. The reagent layer of the younger slide 1 is positioned, and the tip of the spotting tip 53 is positioned above the reagent layer of the slide 1 at the spotting position. Therefore, at the suction position, a predetermined amount of the sample liquid in the sample container 52 is sucked into the spotting tip 53, and then the spotting arm 55 is rotated in the direction of arrow B to move the spotting depth 53 to the spotting position. After positioning, the sample liquid can be dripped onto the reagent layer of the slide 1. After that, this slide 1 is inserted into the empty storage chamber 21 of the incubator 20 and incubated, and the optical reflection density at this time is read by the reading head and the sample F
11 chemical analysis can be performed.

The configuration of the automatic spotting device 50 will be explained based on FIG. 3. This device 50 includes a tip vertical position control means 60 for vertically moving a spotting arm 55 and a spotting tip 53, a container vertical position controlling means 70 for vertically moving a sample container 52, and a container vertical position controlling means 70 for rotating a spotting arm 55 and a spotting tip. 53 between the suction position and the lighting position, and a suction unit 80 that communicates with the chip 53 to aspirate the sample liquid into the chip 53 and discharge the sample liquid from within the chip 53. - Consists of a discharge means 90 and a liquid level detection sensor 100 that detects the liquid level position of the sample liquid in the sample container 52.

The chip vertical position control means 60 is mounted on the main body 10 at a position 1 (
The first pulse motor 61 that has been pulled, the first drive body 63 connected to the rotating shaft of the first pulse motor 61 via a coupling 62, and the female screw 63a formed in the first drive body 63 are engaged. It consists of a first target body 64 having a matching male thread 64b. The first driven body 64 is connected to the protrusion 10 of the main body 10, as shown in FIG.
It has a recessed portion 64a that loosely fits into the recess 64a, and is vertically movable with respect to the main body 10, but is prevented from rotating. Further, a rotating shaft 55a/fi of a spotting arm 55 is connected to the upper part of the first driven body 64 so as to be able to rotate relative to each other. Therefore, the first pulse motor 61
When the driving body 63 is rotated, the first driven body 64 is prevented from rotating, so it engages with the first driven body! - changes, the first driven body 64 is moved up and down, and at this time, the rotating shaft 55a of the spotting arm 55 connected to the first driven body 64 is also moved up and down.

The container vertical position control means 70 includes a second pulse motor 71 attached to the main body 10;
via a coupling 72 to the rotating shaft of the
A female screw 73 formed on the driver 13 and the second driver 73
and a second driven body 74 having a male screw 74b that engages with the second driven body 74. The second driven body 74 has a recess 74a that loosely fits into the protrusion 10a of the main body 10, as shown in FIG. However, rotation is prevented, and
The mounting table 74C on which the sample container 52 is placed is tilted to the right. Therefore, when the second driving body 73 is rotated by the second pulse motor 971, since the rotation of the second fylllJ body 14 is prevented, the screw engagement foot with the second driving body 73 changes, and the second driven body 14 14 is moved up and down, and thereby the sample container 52 placed on the mounting table 74 is moved down.

The chip moving means 80 includes a rotary motor 81 attached to the main body 10, a drive gear 83 connected to the rotating shaft of the rotary motor 81 via a coupling 82, and meshes with the drive gear 83 and is driven. It consists of a driven gear 84 that is movable up and down with respect to the gear 83, and the driven gear 84 is coaxially fixed to the rotating shaft 55a of the spotting arm 55. The rotation shaft 55a of the spotting arm 55 is rotatably supported by the main body 10. Therefore, when the rotation motor 81 is driven to rotate, this rotation is transmitted through the coupling 82, the drive gear 83, and the driven gear 84. It is transmitted to the rotating shaft 55a.

As a result, the spotting arm 55 is rotated around the rotating shaft 55a, and the spotting tip 53 attached to the tip of the spotting arm 55 can be moved between the suction position and the spotting position n. ing.

The suction/discharge means 90 includes a piston drive motor 91 having a cam plate 92 on a rotating shaft, a piston rod 94 connected to the cam plate 92 via a link 93, and a piston rod 94 attached to the tip of the piston rod 94. piston 9
The flexible hose 91 connects a passage 98 formed in the spotting arm 55 and communicating with the space 53a in the spotting tip 53 to a space 96 in the cylinder 95. Piston drive motor 91
The rotational movement of the piston rod 94 is converted into a reciprocating movement of the piston rod 94 by the link 93, and the volume of the cylinder inner space 96 changes due to the reciprocating movement of the piston 95 that occurs.
This is transmitted to the spotting tip internal space 53a via the hose 91 and passage 98, and the sample liquid is sucked into or discharged into the spotting tip internal space IJ 53a.

The liquid level detection sensor 100 optically detects the liquid level position of the sample liquid in the sample container 52, and detects the liquid level position by moving the sample container 52 up and down by the container vertical position control means 70. It looks like this.

The operation of the chemical analyzer δ configured as above will be explained.

First, the reading head moves to a position facing the bottom surface of the cartridge 11, and fog photometry is performed on the lowest chemical analysis slide 1 at the back of the chemical analysis slides 1 stacked in the cartridge 11. When this fog photometry is performed, the chemical analysis slide 1 is pushed out by the pushing lever 12 onto the conveyance/insertion means 40, which has been moved to the position indicated by the chain line X in FIG. 2, and is held there. The conveyance/insertion means 40 moves to the right on the rail 49 to a position below the spotting position (the position indicated by the solid line Y).

Next, based on the spotting method according to the present invention, the sample in the sample container 52 is spotted and supplied by the automatic spotting device 50 onto the reagent layer of the chemical analysis slide 1 that has been transported to the spotting position. However, this spotting is performed as follows.

First, the container vertical position control means 70 raises the @ placing stand 74C, and the liquid level detection sensor 100 moves the sample container 5
Detects the vertical position of the liquid level inside. Next, based on the detected liquid level position, the tip vertical position control means 60 is operated to move the dotting tip 53 attached to the dotting arm 55 in the suction position downward, so that the lower end of the dotting tip 53 is Insert it into the sample solution as shown by the chain line. After this, the suction/discharge means 90 is activated to move the piston 94a,
The sample liquid is sucked into the space 53a of the spotting tip 53 for a predetermined period of time. Note that at this time, the liquid level decreases by suctioning the sample liquid, so this decrease is detected by the liquid level detection sensor 10.
0, the container vertical position control means 70
The loading table 74c may be moved upward to move the sample container 52 upward, thereby keeping the liquid level constant.

When a specified amount of sample liquid is sucked into the space 53a in the spotting tip 53 as described above, the tip vertical position control means 60
The spotting arm 55 is moved upward and the spotting tip 53 is lifted up. Thereafter, the spotting arm 55 is rotated about the rotating shaft 55a by the tip moving means 80, and the spotting tip 53 is moved from the suction position to the spotting position. Next, the spotting tip 53 is placed in the upper and lower tip fffi 1I11.
When it is moved downward by the control means 60 and its tip is located near the upper part of the reagent layer portion 1a of the slide 1, the sample liquid in the spotting tip 53 is slowly discharged by the action of the suction/discharge means 90, and the sample liquid is discharged from the spotting tip 53. At the lower end, an operation as shown in FIG. 6A is formed.

Then, the spotting tip 53 descends as it is, and the reagent is spotted and supplied onto the reagent layer portion 1a of the slide 1 (see FIG. 6B). Note that the distance "h" between the lower end of the spotting tip 53 and the surface of the reagent layer portion 1a at this time is controlled to be always constant. Thereafter, by moving the spotting tip 53 upward, the first spotting of the sample liquid is completed.

Multiple doses of sample liquid can be suctioned and held in the spotting tip 53, and multiple consecutive spottings can be performed while the spotting tip 53 is held at the spotting position. For this purpose, it is necessary to store the spotted chemical analysis slide 1 at the spotting position into a predetermined storage chamber 21, and to position the wood spotted chemical analysis slide 1 at the spotting position. This operation is performed by the conveyance/insertion means 40, and the conveyance/insertion means 40
moves in the lateral direction (arrow C) on the rail 49 to face a predetermined storage chamber 21 of the incubator 920, and after inserting the chemical analysis slide 1 into this storage chamber 21, returns to the chain line in FIG. It moves to the position indicated by X, picks up the unused chemical analysis slide 1 pushed out by the pushing lever 12, and transports it to the spotting position (the position indicated by the solid line Y). In this way, after the unused chemical analysis slide 1 is transported to the spotting position, the ml tan means 60 is placed at the lower tip-F position.
The spotting tip 53 is moved down to perform the second spotting of the sample liquid onto the reagent layer of the chemical analysis slide 1.

In this case, if the transport/insertion means 40 transports the unused chemical analysis slide 1 quickly (if it is carried out, the sample liquid at the tip of the spotting tip 53 exposed to the air will only slightly dry). The second spotting can be performed without clogging the tip, but if the time interval from the first spotting to the second spotting becomes large, the spotting tip 53 This causes the problem of clogging due to drying of the sample liquid at the tip.
If the time interval from the first spotting (previous spotting) to the second spotting (next spotting) is long, the tip moving means 80 moves the spotter arm after the first spotting is completed. 55 to move the spotting depth 53 to the suction position,
2 in FIG. 3 by the chip vertical position control means 60.
As shown by the dotted line, insert the tip of the spotting tip 53 into the sample container 5.
2 to prevent the tip from drying out.

Next, when performing the second spotting, the spotting tip 53 is first lifted above the sample liquid by the tip vertical position control means 60, and the sample liquid in the space 53a of the spotting tip 53 is lifted by the suction/discharge means 90. Discharge completely. After this infiltration, the spotting tip 53 is moved down again to immerse its tip in the sample liquid, and a predetermined amount of the sample liquid is sucked into the space 53a of the spotting tip 53 by the suction/discharge means 90. In this way, even if a dried sample liquid is attached to the tip of the spotter tip 53, it can be removed together with the discharged sample liquid when the sample liquid in the spotter tip 53 is discharged. can effectively prevent clogging.

Roughly speaking, due to changes in the outside temperature a between the first dotting and the second dotting, the inside of the space 53a of the dotting tip 53, the space of the cylinder 95, and the passage 98 may change. Even if the air is expanding or contracting, by discharging the sample liquid once and re-suctioning it, the effects of the expansion and contraction can be eliminated, and the amount of suction and discharge can be accurately controlled. .

The sample liquid thus sucked into the space 53a of the spotter depth 53 is deposited onto the reagent layer of the unused chemical analysis slide 1 at the spotting position when the automatic spotting device 50 is operated in the same way as in the case described above. Supplied on a spot basis. Thereafter, this chemical analysis slide 1 is transferred to a predetermined storage chamber 21 by the transport/insertion means 40.
The temperature is maintained at constant temperature (
incubation), and during this incubation, the irradiation of the illumination light and its reflected optical density are measured by a read head that is moved as needed below the storage chamber;
A chemical analysis of the sample is performed. When these measurements are completed, the chemical analysis slide 1 is transferred to the transport/insertion means 40.
As a result, the receiver is discharged from the storage chamber 21 into the tray 29. Thereafter, by repeating the above operation, the tip of the spotting tip 53 may become clogged.

Spotting can be performed multiple times using the same spotting tip 53, and chemical analysis can be performed automatically and continuously using a large number of chemical analysis slides.

(Effects of the Invention) As explained above, according to the present invention, even when spotting is performed multiple times using the same spotting tip and the spotting interval becomes large, the single spotting tip is By immersing the tip in the sample liquid, drying and clogging of the sample liquid at the tip of the tip can be prevented. In addition, when performing the next spotting, once the sample liquid in the spotting tip has been discharged into the sample container, re-suction is performed to remove the dried sample liquid remaining at the tip of the spotting tip 53. It is possible to eliminate the influence of expansion and contraction of air in the J3 and suction/discharge mechanism, and no clogging occurs.
Moreover, the amount of suction and discharge can be accurately controlled.

[Brief explanation of the drawing]

Fig. 1 is a perspective view showing a chemical analyzer having an automatic spotting device used in the spotting method according to the present invention, Fig. 2 is a plan view showing the chemical analyzer with the cover plate removed, and Fig. 3 is a book. A schematic sectional view showing an example of an automatic spotting device used in the spotting method according to the invention; FIGS. 4 and 5 are sectional views taken along arrows IV-IV and v-■ in FIG. 3; FIG. 6A; and FIG. 6B is a front sectional view showing the operation of spotting the sample liquid from the spotting depth onto the slide. 10...Main body 11...Cartridge 16...Cover plate 20...In 4-Ubeta 40...Transportation/insertion means 50...Automatic point @
Device 53... Spotting tip 55... Spotting arm 60... Tip vertical position e) J'69 means 10.
・Container vertical position control means 80 ・Chip moving means 90 ・Suction/discharge means 100 ・Liquid level detection sensor Figure 1 (Voluntary) Procedures Completed Mr. Seizan Patent Office Commissioner September 1985 2nd, 2nd, Name of the invention, Liquid sample spotting method 3, Relationship with the case of the person making the amendment Patent applicant Address: 210 Nakanuma, Minamiashigara City, Kanagawa Prefecture Name:
Fuji Photo Film Co., Ltd. 4, Agent Address: 5-2-1 Roppongi, Minato-ku, Tokyo 160 6. Number of inventions increased by amendment None 7. Subject of amendment Column 8
, Contents of the correction 1) "Incubator 10" on page 9, line 11 of Book IllI was corrected to "Incubator 20." 2) 15th true 3rd line

Claims (1)

[Claims] The tip of the spotting tip is immersed in the sample liquid placed in the sample container, a predetermined amount of sample liquid is sucked and held from the tip into the spotting tip, and then the sample liquid in the spotting tip is A method of spotting on a reagent layer of a chemical analysis slide, in which the spotting is performed multiple times using the same spotting tip, in which the spotting is performed multiple times from the previous spotting to the next spotting. If the time interval between spotting is longer than a certain value, the tip of the spotting tip is dipped into the sample solution in the sample container after the previous spotting is completed, and for the next spotting, the tip is removed from the sample solution. After drawing out the sample liquid left in the spotting tip and discharging it into the sample container, the tip is immersed in the sample liquid again and a predetermined amount of sample liquid is poured from the tip into the spotting tip. 1. A method for spotting a liquid sample, comprising: suctioning and holding the sample liquid in the spotting tip, and then spotting the sample liquid in the spotting tip onto a reagent layer of a chemical analysis slide.