JPS6252448A - Method for measuring concentration in electrophoretic apparatus - Google Patents

Abstract

PURPOSE:To attain an increase in treatment speed by shortening a treatment time, by applying staining and decolorization treatments to a support after electrophoretic treatment and subsequently measuring fractional concn. in such a state that the treated support has been immersed in a decoloring solution. CONSTITUTION:A support A after electrophoretic treatment is sent into a liquid tank 11 through an inlet side passage (a) by rollers 20a, 20b and subsequently sent into the lower part of the liquid tank 11 while being held between rollers 22a, 22b to be immersed in a staining solution. The staining solution is discharged from the liquid tank 11 and a decoloring solution is introduced into said tank 11 to decolor the support A. Further, a required amount of the decoloring solution is introduced into the liquid tank 11 to fill the same so as to reach the vicinity of the lower parts of the rollers 20a, 20b and, when the support A is sent to a predetermined position, said support A is held between the rollers 20a, 20b to be stopped and held at that position. Subsequently, a feed screw shaft is rotationally driven by a feed motor. A light transmitting/receiving part support stand is guided on a guide shaft through a feed nut and linearly fed to a predetermined direction and fractional concn. is measured along the film surface of the support A.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application This invention relates to a method for measuring concentration in an electrophoresis apparatus.

BACKGROUND OF THE INVENTION Electrophoresis has been used to test serum proteins in conventional clinical laboratories in medical institutions. In this electrophoresis method, as is well known, the serum to be tested is coated on a support such as a cellulose acetate membrane, and then electricity is applied to form an image of the serum. This energized sample is stained with a staining solution,
Furthermore, after decolorizing parts other than serum, the fractional concentration is measured using a densitometer.

By the way, it would be extremely inefficient to perform these steps manually, and testing using electrophoresis requires a great deal of skill. Problems may occur, such as incorrect results.

Based on the above, we automated the various steps of this electrophoresis method,
In addition to improving efficiency, it also eliminates variations between inspections.
Efforts are being made to put into practical use automated devices that can obtain good test results.

Problems to be Solved by the Invention However, in the practical devices that have been proposed so far, the support that has been subjected to electrophoresis is dyed and decolorized, and then further dried, and the dried material is made transparent. The configuration was such that the concentration was measured only after the

This drying process is easily affected by the outside temperature and humidity, and it often happens that complete transparency cannot be obtained due to insufficient drying. In addition, overdrying may cause wrinkles on the support, which has the problem of adversely affecting measurement results. Furthermore, since the two steps of drying and transparency are indispensable, the number of processing steps increases and the processing time is long, making it impossible to achieve rapid processing.

This invention was proposed to solve the above-mentioned conventional problems, and aims to eliminate the need for the two steps of drying and transparency, shorten the processing time of the electrophoresis step, and speed up the processing. The purpose is to

In order to achieve an objective that is more than just a means to solve the problem, the present invention aims to dye and decolorize a support that has undergone electrophoresis, and then immerse it in a decolorizing solution to determine the density of the electrophoretic pattern. I started taking measurements.

Function According to the above structure, the density of the dyed/decolorized support is immediately measured after being immersed in the decolorization solution after the electrophoresis treatment.

Therefore, the two steps of drying treatment and clarifying treatment are omitted, and the density is measured immediately after the decolorization treatment. by this,
Double edges that adversely affect measurement results, such as insufficient drying, resulting insufficient transparency, and wrinkles on the support due to overdrying, are eliminated.

Embodiments Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.

In FIGS. 1 and 2, 10 is a casing, inside which a pair of block bodies are butted together and integrated.
A decolorizing liquid tank 11 is provided. The dyeing/decolorizing liquid tank 11 has a width wider than the width of the support in the front and back directions of the paper in FIG. It is designed to the extent that it is possible to measure the fractional concentration on the support. A drain port 12 is provided from the bottom of the liquid tank 11 to the side wall. The opening and closing of the drain port 12 is controlled by, for example, a solenoid valve.

The upper surface of the casing 10 above the liquid tank 11 is open, and a top cover 13a is removably attached thereto.

Between this upper cover 12 and the upper surface of the liquid tank, there is a passageway on the inlet side of the support (
A) and an exit passage (b) are formed. Passage (a)
, (b) are inclined downward from the outside of the liquid tank toward the center of the inside. The support is fed into the liquid tank 11 through the inlet side passage (a), and is discharged from the liquid tank 11 to the outside through the outlet side passage (b). The lower part of the right side wall of the liquid tank 11 shown in FIG. 1 is open, and a lower lid 13b is sealed therein and screwed into the lower part.

A pair of feed rollers 14 and 15 are placed at positions facing the entrance end of the entrance passageway (a) at intervals in the direction of the front and back of the paper in FIG.
is provided. The support body that has undergone the electrophoresis treatment in the previous step is fed by feed rollers 14 and 15 and sent to the entrance side passage (a). Furthermore, a pair of support discharge rollers 16 and 17 are disposed at a position facing the exit end of the exit side passage (a), spaced apart from each other in the front and back directions of the paper in FIG.

The support that has been dyed and decolorized inside the liquid tank 11 and whose density has been measured passes through the exit side passage (b) and is delivered to the discharge rollers 16 and 17.
The waste is fed to the outside of the device and discharged.

Concave portions 18a, 18b and concave portions 19a, 19 are provided on both sides of the upper part of the liquid tank 11 and on both sides of the middle part spaced a certain distance below.
b is formed.

In the recesses 18a and 18b, a pair of first rollers 20a and 20b are supported by shafts 21a and 21b, respectively, and spaced from each other in the left-right direction in FIG. Similarly, each pair of second rollers 22a and 22b are attached to the shaft 28a in the recesses 19a and 19b.
, 28b, and are spaced apart from each other in the left-right direction in FIG. 2, and are provided in pairs facing each other.

In addition, in the following explanation, the roller 20a122a will be explained as a fixed roller, but it will be explained as the roller 20b.
, 22b.

On one side of the liquid tank 11, a support feed motor 24 is fixed to a support plate 25 and arranged.

A shaft 21a supporting the first a-220a is connected to the shaft via a coupling 26. The other end of the shaft 21a protrudes to the other side of the liquid tank 11, and a pulley 2 is attached to the end of the shaft 21a.
7 is installed. A belt 29 is passed between this pulley 27 and a pulley 28 attached to the end of the pongee 28a.

The rotational force of the motor 24 is transmitted to the first shaft 2 through the shaft 21a.
At the same time, it is transmitted to the 20th roller 22a through the belt 29 and the shaft 23a. Therefore, the first and second rollers 20a.

22a are rotationally driven in the same direction and synchronously by one drive of the motor 24. And motor 24 is positive.

When reversely driven, the first and second rollers 20a.

22a is driven to rotate forward and backward in the direction in which the support body is sent into the liquid tank 11 and in the direction in which it is sent out, respectively.

first rollers 20a, 20b and second roller 22a.

The opposite side wall of the liquid tank 11 between the liquid tank 22b and the support 22b is hollowed out in a rectangular shape with a size large enough to cover the area of the migration pattern formed on the support. Transparent or light-transmissive window plates 32.88 made of glass, resin, etc. are attached to the liquid tank walls of the opening windows ao and si thus cut out.

Two mutually parallel guide shafts 34, 34 are arranged on one side of the liquid tank 11. guide shirt l-34,
A bearing plate 35 is fixed to the free end of 34. A light projecting section and a light receiving section constituting the concentration measuring unit are mounted on the guide shaft 84.34. The light projecting portion is guided by guide shafts 84 and 84 and is connected to the window plate 3 described above.
2 and 33, and linearly fed along the membrane surface of the support. The drive mechanism is constructed as follows.

- A light emitting/receiving section feed motor 36 is installed on the inner wall of the casing 10. A feed screw shaft 37 is supported between the casing 10 and the bearing plate 35 so as to be rotatable in forward and reverse directions.

A belt 40 is passed between a pulley 38 provided at the shaft end of the feed motor 36 and a pulley 39 provided at the shaft end of the feed screw shaft 37 to transmit the driving force of the motor to the feed screw shaft 37. It has become.

A feed nut 42 fixed to a stand 41 is screwed into the nut receiver of the feed screw shaft 371C. This nut holder is stand 41
One end side of a light emitting/receiving unit support stand 43 is supported on the top.

The light projecting section 50 and the light receiving section 60 are disposed on the support base 42 so that their optical axes are coaxial and face each other with the window plates 82 and 88 in between.

The light projection unit 50 includes an optical fiber 51 that guides light from a light source (not shown) inside, a mirror 52 that reflects the incident light by 90 degrees, a lens 53 disposed in front of the mirror 52, and a window in front of the optical fiber 51. It consists of a plate 82 and a slit 54 provided facing each other.

The light coming out through the slit 54 is transmitted to the window plate 82. The light is received and detected by the light receiving section 60 through 'a8, and after being photoelectrically converted, it becomes an electric signal corresponding to the fractional concentration of the support, and density data is extracted through the signal processing system.

In the above configuration, the support A that has undergone the electrophoresis process is sent into the liquid tank 11 through the entrance side passage (A) by the rotation of the first rollers 20a and 2ob. At this time, there are second rollers 22a and 22b in the liquid tank 11.
It is filled with staining solution to approximately the same height as .

The support A sent in as described above is transferred to the second roller 22
It is sandwiched between a and 22b, and as it rotates, it is sent to the lower part of the liquid tank 11 and immersed in the dyeing liquid. Then, a sample staining process is performed.

When the dyeing process is finished, the dyeing liquid is discharged from the liquid tank 11, and at the same time, the decolorizing liquid is transferred to the second roller 22a.

It can be inserted to approximately the same height as 22b. Then, the support A in the tank 11 is immersed in this decolorizing solution, and the portion other than the sample serum is decolorized.

When the decoloring process is completed, a required amount of decolorizing liquid is further poured into the liquid tank 11, and the first roller 20a is moved.

20b is filled to near the bottom. Next, the support body A is sent upward by the rotational drive of the second coolers 22a and 22b. When the support body is sent to a predetermined position, the first
．． It is held between the second rollers 20a, 20b and 22a, 22b, and is stopped and held at that position. In this state, the support A is kept immersed in the decolorizing solution. Next, by driving the feed motor 36, the feed screw shaft 37 is rotated in a predetermined direction and made to vibrate.

Now, the projector/receiver support base 43 is moved via the feed nut 42 to the guide shaft a4. It is guided by a4 and linearly fed in a predetermined direction. As a result, the light receiving section 50.6
0 is sent along the membrane surface of the support A through the window plates 82 and 8B in the direction of measuring the fractional concentration.

Then, electrical signals corresponding to the density of the electrophoretic pattern formed on the support A are sequentially and continuously taken out from the light receiving section 60, and density data is measured and a density map is drawn at the same time. In this manner, after the decolorization treatment of support A, the density is immediately measured while the support is immersed in the decolorization solution.

As described above, after decolorizing the support, measurements are carried out immediately while the support remains opaque without drying or clarifying treatment. In this case, the relationship between the degree of attenuation and the absorbance becomes a curve as shown in FIG. Therefore, the measured light attenuation is converted into absorbance using the following correction formula.

dEt E=dEt71agP(1-10) (where, E: absorbance, dEt2 attenuation, log
P=intercept shown in FIG. 3) Thus, when a concentration measurement is made, the first. second roller 2
The support body A is sent upward by the rotation of 0a, 20b and 22a, 22b, and is carried through the exit side passage (b) to the support body discharge rollers 16, 17, and is carried by both rollers 16, 1.
7 and is fed to the outside.

As described above, dyeing, decolorization, and density measurement are carried out on the support A.
This is done in series while the water is still immersed in the same liquid tank.

In the examples, a so-called transmission measurement method was described in which the light from the light projecting section is transmitted through the support and detected by the light receiving section. Of course, the present invention is also applicable to a reflection measuring method in which the reflected light is received by a light receiving section.

As explained in detail, the method of this invention does not require a clarifying liquid and can omit the two steps of drying and clarifying, so the processing time is shortened compared to the conventional method. , it is possible to achieve rapid processing of measurements. In addition, it is possible to eliminate factors that affect measurement, such as unevenness during transparency due to insufficient drying and wrinkles due to overdrying, making it possible to perform good and accurate concentration measurements.

[Brief explanation of drawings]

Fig. 1 shows a specific example of the apparatus used in carrying out the method of the present invention. FIG. 3 is a characteristic diagram showing the relationship between light attenuation and absorbance in FIG. A・・・・・・・・・・・・・・・Support, 11
・・・・・・・・・・・・・・・Liquid tank, 30.81...
...Window part, 20a, 20b... First roller, 22a, 22b
... Second roller, Figure 1

Claims (1)

[Claims] (1) A method for measuring concentration in an electrophoresis apparatus, characterized in that after dyeing and decolorizing a support that has been subjected to electrophoresis treatment, the fractional concentration is measured while immersing it in a decolorizing solution.