JPS6258457B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a fully automatic cassette electrophoresis apparatus.

In electrophoretic analysis, samples such as serum proteins are
When a DC current is applied to both ends of a support containing a PH buffer, the buffer ions and charged molecules or atoms migrate in the direction of the electrode opposite to their load, and after a certain period of time, the migration speeds differ. Since the fractions are arranged on a support in order and separated, the amount of each fraction separated is quantified based on the difference in separation speed. For this purpose, each sample is coated on a thin film moistened with a buffer solution, electrophoresed in an electrophoresis tank, fixed and stained, and the concentration of each substance fractionated on the electrophoresis membrane is determined. It is for reading.

However, the electrophoresis described above is sensitive to the effects of uneven quality of the thin film that is the sample support, expansion and contraction of the thin film during drying, wetting, re-drying, and transparency, as well as ambient temperature and humidity. The above-mentioned sample processing requires a considerable degree of skill, individual differences are inevitable, and there are disadvantages such as the need for a lot of time and labor, and the inconvenience of sample storage after measurement.

In order to overcome this drawback, an electrophoresis apparatus capable of performing these operations automatically was proposed in Japanese Patent Publication No. 55-6190, and some of these devices have been put into practical use.
The electrophoresis device described above uses a long strip-shaped membrane such as a cellulose acetate membrane, and wets the strip-shaped membrane with a buffer solution.
The device is continuously transported to locations such as sample application, electrophoresis, staining, decolorization, and concentration measurement. However, in such a method, the belt-like membrane itself is a thin film that is in an extremely weakly wet state, and has especially uneven elasticity, so that the belt-like membrane cannot be smoothly transported through each stage of the device. Failure of the equipment or even cutting of the strip membrane may occur, requiring the entire operation to be stopped, which poses a major obstacle to automation. Furthermore, the damage caused by the loss of valuable patient serum is extremely large.

Furthermore, for continuous operation, it is necessary to transport the strip membrane at a constant speed, and therefore the size of equipment for each process must be designed based on this. In particular, electrophoresis takes a long time (approximately 30 to 50 minutes), so when processing a large number of samples, it is necessary to use a larger electrophoresis tank, or to use two or more electrophoresis tanks because the residence time of the strip membrane in the electrophoresis tank is long. This requires a complicated mechanism such as the provision of a plurality of electrophoresis tanks. Also, in this case, the disadvantages of the above-mentioned strip membrane processing increase the accuracy of the device and the difficulty of remediation against failures. In order to avoid the above fatal drawback, a method has actually been proposed in which the above-mentioned membrane strips are cut to the required length and sequentially transferred to each process, but this method requires a lot of effort to clamp the membrane strips. In transport means such as rollers, the unsmoothness of operation caused by changes in the wet (or dry) membrane strip remains unresolved, and in order to eliminate this, many guide means are provided. It requires a complicated mechanism, and has the disadvantage of being prone to failure.

An object of the present invention is to provide a fully automated cassette-type electrophoresis apparatus that can smoothly and reliably perform electrophoretic analysis of serum proteins and the like.

The automatic electrophoresis apparatus according to the present invention is an apparatus characterized by having the following components (a) to (k).

(a) A sample plate stocker in which sample plates for storing multiple samples such as serum proteins are stacked vertically.

(b) Cut the electrophoretic membrane into a size that allows for coating and measuring the sample in one sample dish, fix this in a cassette (consisting of a rigid upper and lower frame), and stack the cassettes vertically. A case for storage.

(c) From the bottom of the cassette stocker, place the cassette 1
Wetting means that takes out the electrophoretic membrane one by one and immerses it in a buffer solution to moisten the electrophoretic membrane with the buffer solution.

(d) A sample supply means that takes out sample plates one by one from below the sample plate stocker and applies the sample to the electrophoretic membrane after wetting.

(e) Guided by endless rails laid on approximately the same horizontal plane, latched to an endless chain laid along the rails, moved by the chain, gripping and releasing means for the cassette, and upper and lower parts of the gripping cassette. A handler having a moving means, which receives and grips a cassette from a sample supply means, transfers it to an electrophoresis tank, a staining tank, a decolorization tank, and a drying means in this order, and then returns to the cassette receiving position.

(f) Multiple migration tanks installed along the rail. The electrophoresis tank is used alternately, and the lid is opened and closed and the buffer solution is exchanged in response to the transfer, gripping, and regripping of the cassette by the handler.

(g) A dyeing tank and a decolorizing tank installed along the rail and spraying a dyeing liquid and a decolorizing liquid onto the cassettes using spray nozzles, respectively. The lids of the staining tank and the decolorizing tank are opened and closed, and spraying is performed in response to loading, gripping and regripping of cassettes by the handler.

(h) Drying means provided along the rail to dry the cassettes after decolorization. When a cassette is carried in and released from the grip by a handler, the drying means closes the lid, dries the cassette, and transfers the dried cassette to the measuring means.

(i) Measuring means consisting of a decalin roller section that transparentizes the electrophoretic membrane of the cassette sent from the drying means, and a densitometer section that quantifies the amount of each fraction on the electrophoretic membrane of the cassette that has passed through the decalin roller section.

(j) Control means for controlling the transportation of cassettes and sample dishes to each means, the operation of each means, and the operation of the corresponding handlers.

(k) Means for detecting, confirming, and recording the cassette code and sample plate code.

One of the main features of the device of the present invention is that the electrophoretic membrane for applying a plurality of samples is fixed between rigid upper and lower frames (the central part other than the four sides is cut out) to form a cassette. The electrophoretic analysis process is performed completely automatically on a cassette basis.

Another main feature of the apparatus of the present invention is that the cassette is sequentially gripped and transported from the sample supply means to the electrophoresis tank, staining tank, decolorization tank, and drying means in this order by a handler that moves along an endless track. , being transferred while being accompanied by operations such as gripping, releasing, and regripping.

Yet another feature of the apparatus of the present invention is that a plurality of, for example four, electrophoresis tanks are provided, and by using the electrophoresis tanks alternately, the time required for each step is equalized, and the overall time required is is shortened.

Other features of the device of the invention will become apparent from the detailed description provided below.

Hereinafter, the apparatus of the present invention will be described in detail based on the attached configuration diagram (FIG. 1) showing one embodiment.

In the figure, 10 is a sample plate stocker, 20 is a cassette stocker, 30 is a wetting means, 40 is a sample supply means (for example, an applicator), 50 is a handler, 60 is an electrophoresis tank, 70A is a staining tank, and 70B is a decolorizing tank. , 80 is a drying means, 90A is a decalin roller portion of the measuring means 90, and 90B is a densitometer portion. Although the apparatus of the present invention has a separate control means, it is not shown because it is provided for each process.

Conventionally, in order to improve testing efficiency, a plurality of samples, such as serum samples, for example, 20 samples, are stored separately in a single serum dish. 11 is a serum dish,
The serum dishes 11 are housed in a vertically stacked manner in the sample dish stocker 10. The serum dishes 11 are delivered one by one to the sample supply means 40 according to a command from the control means. During the delivery, the code on the serum dish 11 is read by the light receiving element, and the code is fed back to the control means and stored to prevent misidentification of the sample. Further, a counter for the number of dishes is provided, and together with a sensor for checking the number of dishes provided in the stocker 10, the amount of stock in the stocker 10 is checked. The serum dish 11 is fed by a commonly used method, such as moving a feed plate using a rack and pinion.

A cassette stocker 20 stores cassettes 21 stacked vertically. The cassette 21 is made by cutting a migration membrane such as a cellulose acetate membrane into a size capable of coating and measuring the sample in one sample dish 11, and fixing it between upper and lower rigid frames. The cassette 21 is also taken out from the stocker 20 and checked in substantially the same manner as the sample dish 11.

The cassette 21 taken out from the cassette stocker 20 is immersed in the buffer solution tank 30 by a means such as a clamping roll guided by a guide rail, and the electrophoretic membrane is wetted with the buffer solution. The cassette 21 that has been immersed in the buffer solution waits for application of serum by the applicator of the sample supply means 40.

At the sample supply means 40, the applicator 40 of the sample supply means sucks up serum from the serum dish 11, reaches the cassette 21, and after applying the serum to the cassette 21, washes the tip (blood collection part). The tank is indicated by 41) and cleaning (the cleaning means is indicated by 42).
Indicated by ), then perform the next blood collection. The used serum dish 11 is received by a pedestal 43 having a balance spring and stored in a used dish stocker 44.

The sample coated cassette 21 is transported by elevator or other suitable means to a suitable position for feeding into the handler 50.

51 indicates a handler moving conveyor line. The handler moving conveyor line 51 is made up of an endless rail that guides the handler 50 and is laid on the same horizontal plane, and a chain that is laid along the rail and rotated by a drive source controlled by a control means. 50 is engaged with the chain.

An embodiment of handler 50 is shown in FIG. Three arms extend from the main body 52 of the handler 50, and rollers 53 attached to the two arms connect to the two endless rails 51.
The handler 50 is guided by the handler A. Moreover, since one arm is locked to the chain 51B, the handler 50 moves along the rail 51A as the chain 51B rotates.

On the opposite side of the arm of the handler body 52, a slide bar 54 is provided parallel to and perpendicular to the body 52 with a slight spacing therebetween. A slide plate 55 is fitted into the slide bar 54,
The slide plate 55 is moved up and down along the slide bar 54 by driving a slide plate moving lever 56 attached to the slide plate 55 from the outside. The slide plate 55 is always fixed by a fixed ball catch 57, and when the slide plate moving lever 56 is driven, it slides out of the ball catch 57.

Further, a fixed arm 58A is fixed to the slide plate 55, and an opening/closing arm 58 is fixed to the fixed arm 58A.
B is pivoted, and the fixed arm 58A and the opening/closing arm 5
8B can hold the cassette 21. A pinion gear 58C pivotally attached to the fixed arm 58A meshes with the teeth of the opening/closing arm 58B, and also meshes with an arm rotation lever 58D pivotally attached to the slide plate moving arm 56. Therefore, when the arm rotation lever 58D is driven from the outside and rotated in the direction of the arrow A, the space between the fixed arm 58A and the opening/closing arm 58B opens, allowing the cassette 21 to be removed. Reference numeral 59 denotes a torsion spring for keeping the arms 58A and 58B in a state of gripping the cassette when the rotary lever 58D is not driven.

The handler 50 receives the cassette 21 from the sample supply means 40 and transfers the cassette 21 to a migration tank 60 provided along a conveyor line 51.
In the illustrated embodiment, there are four migration tanks (60A,
60B, 60C, 60D) are provided. However, the number of electrophoresis tanks is determined as appropriate, particularly taking into consideration the electrophoresis time of the sample and the required number of specimens to be tested, that is, the required testing speed.

Each migration tank 60 has a lid that opens and closes automatically by moving up and down. When the handler 50 grips the cassette 21, the lid of the first migration tank 60A opens, the handler 50 stores the cassette 21, and when the handler 50 returns to its original position, the lid closes and migration begins.
In this way, the second, third and fourth tanks, 60
B, 60C, and 60D perform electrophoresis in the same manner. A direct current is applied to the electrophoresis tank 60, and the protein molecules of the sample on the electrophoresis membrane are separated based on the difference in migration speed. For this purpose, DC electrodes are connected to both sides of the electrophoretic membrane.

After a while, the electrophoresis in the first tank is finished, the lid is opened, the handler 50 comes and grasps the cassette 21, and the cassette 21 is carried to the next staining tank 70A, and the lid is closed. 2nd tank 60B
Similarly, when the electrophoresis is completed, the cassette is sent to the next step. In order to perform electrophoresis again in each electrophoresis tank 60,
When the lid is opened, the buffer solution in the left and right sections of the tank is exchanged via various devices such as attached piping and separate tanks.

Next, along the rail, a dyeing tank 70A and a decoloring tank 70
B is provided. In the dyeing tank 70A, the staining solution is sprayed from the bottom upwards with one nozzle, and in the decolorizing tank 70B, the decolorizing solution is sprayed upwards from the bottom with one nozzle and from the top downwards with three nozzles.
This spraying method can process in an extremely short time compared to the previous method, which uses a drum rotating system, and stains and decolors while rotating an electrophoretic membrane fixed around the drum in a tank. be. Dyeing tank 70
Although the decolorization tank A and the decolorization tank 70B are also covered, the opening and closing of the lids and the operation of the handler 50 are generally the same as in the migration tank 60. However, the handler 50 is on standby during dyeing or bleaching, and when the process is finished, the cassette 2
1 and transfer it to the next process. By adopting this method, the related equipment will not be corroded by the acidity of the dyeing solution and decolorizing solution, unlike conventional machines.
Processing time can also be significantly reduced.

Next, a drying means 80 is provided along the rail 51A. In this embodiment, a hot plate and a fan are used to heat the aluminum plate with a strip heater. The electrophoretic membrane part of the cassette is placed on a hot plate and heated, and a lid equipped with a hot air fan is closed to dry the cassette with hot air. The operation of the handler 50 and the lid and other elements of the drying means 80 is generally the same as in the previous step. However, the handler 50 which has released the cassette 21 by the drying means 80 is returned to the receiving position of the new cassette 21. Further, the cassette 21 that has been dried is pushed by a transfer plate and transferred to the measuring means 90.

The measuring means 90 consists of a decalin roller section 90A and a concentration meter section 90B. The cassette 21 sent from the drying means 80 is placed on a sample carrier and moved to the decalin roller portion 90A, and the sample carrier is moved while supplying decalin to the rollers to make the electrophoretic film transparent. Next, the cassette 21 is sent to the densitometer section 90B and its density is measured. After the measurement is completed, the sample carrier returns to its initial position and ejects the measured cassette 21. Next, the sample carrier moves to the decalin roller area, uses the decalin roller to clean the area on which the electrophoretic membrane is placed, returns to its original position, and waits until the next cassette arrives.

Although some explanations are omitted in the description of the device of the embodiment, the device of the present invention is equipped with a control means, whose operation is programmed, and which decodes the program and controls each device according to the program. means for controlling the operation of the apparatus of the invention and for detecting and verifying the cassette code and the sample pan code.

Since the apparatus of the present invention is constructed and operates as described above, it can achieve the following superior effects compared to conventional automatic electrophoresis apparatuses.

(1) Since the electrophoretic membrane is formed into a cassette, the operation of the device does not become unsmooth due to expansion and contraction of the membrane, which is the case with strip-shaped electrophoretic membranes.

(2) By considering the electrophoresis time and other processing times, it is possible to determine the number of electrophoresis tanks that correspond to the required number of samples to be processed.

(3) Since the cassettes are carried in and out of each process by a handler, there are almost no failures, unlike the case of moving the strip membrane, and programming is easy and rational automatic control can be performed.

(4) With band-shaped membranes, codes can only be attached by drilling holes, etc., but in the apparatus of the present invention, clear codes can be attached to the cassettes, so there are fewer operations for attaching codes during the process, and there are fewer errors.

[Brief explanation of the drawing]

Fig. 1 is a block diagram of an embodiment of the device of the present invention;
The figure is a side view of an embodiment of the handler of the device of the invention. 10... Sample plate stocker, 11... Sample plate, 2
0...Cassette stock, 21...Cassette, 3
0... Wetting means, 40... Sample supplying means, 50...
...Handler, 51...Handler moving conveyor line, 51A...Endless rail, 51B...Endless chain, 58A...Fixed arm, 58B...Opening/closing arm, 60...Migration tank, 70A...Staining tank, 70
B... Decolorization tank, 80... Drying means, 90... Measuring means, 90A... Decalin roller section, 90B...
Concentration meter section.

Claims (1)

[Scope of Claims] 1. A cassette-type fully automatic electrophoresis apparatus characterized by comprising the following items (a) to (k). (a) A sample plate stocker that stores multiple samples such as serum proteins by stacking them vertically; (b) A sample plate stocker that stores multiple samples such as serum proteins by stacking them vertically; (b) A sample plate stocker that stores multiple samples such as serum proteins, etc.; (c) A cassette stocker in which the cassettes are cut into strips and fixed on a rigid frame to form a cassette, and the cassettes are stacked vertically for storage. (d) sample supply means that takes out the sample plates one by one from the sample plate stocker and applies the sample to the cassette from the wetting means; (e) the sample plates laid on approximately the same horizontal plane; It is guided by an endless rail, is latched to an endless chain laid along the rail, is moved by the chain, has means for gripping and releasing the cassette, and means for moving the gripped cassette up and down, and has a means for moving the gripped cassette up and down. A handler that receives and grips the cassette, transfers it to the electrophoresis tank, dyeing tank, decolorization tank, and drying means in this order, and returns to the cassette receiving position again; (f) a plurality of migration tanks provided along the rail; The tank is used alternately, and includes an electrophoresis device that opens and closes the lid and exchanges the buffer solution in response to transport, grip release, and re-gripping of the cassette by the handler; and a dyeing tank and a decolorizing tank in which a decolorizing solution is sprayed by a spray nozzle; a dyeing and decolorizing means for opening and closing a lid and spraying in response to loading, gripping release, and regripping of a cassette by a handler; h) Drying means installed along the rail to dry the cassettes after decolorization; when the cassettes are carried in and released by the handler, the drying means closes the lid, dries the cassettes, and measures the cassettes after drying. (i) A decalin roller section that transparentizes the electrophoretic membrane of the cassette sent from the drying means, and a densitometer section that quantifies the amount of each fraction on the electrophoretic membrane of the cassette that has passed through this section. (j) a control means for controlling the transport of the cassette and sample pan to each means, the operation of each means and the operation of the corresponding handler; and (k) detecting the cassette code and the sample plate code; A means of verifying and recording this.