JPS5933217B2 - densitometer - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an electrophoresis apparatus, particularly a densitometer used in an electrophoresis apparatus for analyzing serum proteins, using a cellulose acetate membrane as a support for a buffer solution.

Cellulose acetate electrophoresis equipment has low adsorption of specimens and dyes, minimal sample adsorption, so there is little sample loss, and even minute amounts of specimen can be analyzed, and there is little tailing during electrophoresis, so separation is clear. It has advantages such as being advantageous for quantitative determination.

The applicant has developed an automatic electrophoresis device that can simultaneously perform such cellulose acetate electrophoresis on a large number of specimens. This automatic electrophoresis device has a part that cuts a rolled support into a predetermined length;
A part that wets the support with a buffer solution, a part that applies a serum sample to the support, an electrophoresis part that fractionates the serum applied to the support, and a part that stains the support after electrophoresis. and a densitometer part for decolorizing the dyed support and measuring the density. Conventionally, in a densitometer used in such an electrophoresis device, a support on which a separated image has been formed by electrophoresis is sandwiched between two transparent plates, or is attached to one transparent plate, and then the density is measured. was.

In both cases the support must be transparent. This can be done by sandwiching the support, which has been made transparent by soaking it in a clarifying liquid in advance, between two transparent plates, or pasting it on one transparent plate. Methods have been considered to do this, but in any case, air bubbles tend to enter, and the clarifying liquid tends to spill around, making the process complicated. The applicant has proposed a densitometer in JP-A-50-132993, which eliminates the above-mentioned drawbacks and allows photometry to be performed without first making the support transparent after electrophoresis and dyeing.

In this densitometer, the support is placed in a nearly sealed container filled with a clarifying liquid to make it transparent, and the light is measured there. However, the light source and light receiving part that make up the photometry section are placed above and below the container and away from the container. It is arranged as follows. That is, the light emitted from the light source is made to enter the container through the bottom surface of the container, and the light transmitted through the support is made to enter the light receiving section through the top surface of the container. In such a configuration, since the light passes through the interface between the container and the air twice, it is susceptible to reflection from the container surface, resulting in a reduction in photometry accuracy. In addition, since the upper and lower walls of the container are placed opposite each other with a narrow interval, air bubbles that enter when the support is immersed in the clarifying liquid remain, which adversely affects concentration measurement. There is also. An object of the present invention is to provide a densitometer that can make a support on which an image has been formed transparent by a simple operation and can measure density with high accuracy.

The densitometer of the present invention has a container with an open top for containing a clarifying solution, and a light source and a light receiving part arranged oppositely above and below this container, and the lower end of the light source or the light receiving part arranged above the container is made transparent. By changing the relative position of the clarifying solution solution and the measuring section while the support of the object to be measured remains in the container containing the clarifying solution. , is characterized in that it is configured to scan each density pattern on the support and to perform feeding between each density pattern.

The present invention will be described in detail below with reference to the drawings.

FIGS. 1 to 3 show the configuration of a sample to be subjected to a densitometer using a conventional method. With conventional densitometers,
A sample was made by sandwiching the transparent support between two transparent plates or pasting it on a single transparent plate, and the sample was placed in the measuring section of a densitometer to measure the concentration. . Figure 1 shows how a support is sandwiched between two transparent plates (glass etc.). Support 1, which has been made transparent by soaking it in a clarifying liquid in advance, is placed on top of transparent plate 2. A transparent plate 3 is placed on top of the sample 4, the cross section of which is shown in FIG.
That is, a transparent support sandwiched between two transparent plates is made. Alternatively, instead of making the support 1 transparent in advance, a method may be considered in which a required amount of clarifying liquid is poured onto the transparent plate 2, the support 1 is placed on top of the transparent plate 2, and then the transparent plate 3 is placed on top of the transparent plate 3.
In either method, the transparent plates 2 and 3 are removed during the process of making sample 4.
Air bubbles tend to remain between the two, interfering with measurement, and removing these air bubbles is a very difficult task. Furthermore, when preparing sample 4 using these methods, it is difficult to determine the amount of clarifying liquid, and if there is too much, it may protrude outside the transparent plates 2 and 3 and stain the transparent plates and the sample stand of the densitometer. If the amount is too small, it may cause bubbles to form. In order to solve these problems, as shown in Fig. 3, the upper transparent plate is replaced with a flexible material film 5 such as a thin transparent plastic film to make it easier to remove air bubbles. Alternatively, the transparent plate on the top was omitted.

However, in any case, the complexity of the work could not be resolved. FIG. 4 shows the overall configuration of a densitometer according to the present invention, which solves the above-mentioned operational complexity.

FIG. 4a shows a cross section, and FIG. 4b shows a longitudinal section. In the present invention, instead of preparing a sample with a previously transparent support and placing it at the measurement position of a densitometer as in the conventional case, the support transparent part is integrated with the density measurement part. That is, a frame-shaped or transparent mounting table 6 is provided so as to be movable in the direction of arrow 7 and in the direction perpendicular to the plane of FIG. 4a, that is, in the direction of arrow 8. It is made of a transparent substance and a transparent liquid solution container 10 is placed thereon. Furthermore, this clearing liquid solution 10
An illumination section 11 and a light receiving section 12 are arranged at positions facing each other with the bottom surface 9 interposed therebetween. The lighting section 11 includes a light source lamp 11A,
A condenser lens 11B and a filter 11C are provided. The light receiving unit 12 placed at the top is aligned with the axis 1 as shown in Fig. 4b.
The container 10 is attached via an arm 14 which is rotatable around the container 10, and the light receiving part 12 is configured to be detachable from the upper surface of the container 10 as necessary. The inside of the container 10, which is open at the top, is filled with a transparentizing liquid 15 until at least the light receiving part 12 is immersed therein.

In the present invention, the lower end of the light receiving section 12 is connected to the transparent liquid 1 in this way.
5, it is possible to completely eliminate the influence of diffused reflection on the liquid surface, making it possible to perform highly accurate concentration measurements. Moreover, since the upper part of the container 10 is open, air bubbles are quickly dissipated and do not affect the measurement. Next, the light receiving section 12 is raised and the support body is placed into the container 10. The support to be measured has a length a from the support tip 16 (FIG. 1) to the first partial image center line 17-1 (FIG. 1), and a length a of each partial image center line 17-1 to 17-n ( The pitch P between (Fig. 1) is regulated to be constant. When using such a support (not made transparent) 1, the support tip 16 is shown in FIG. 4a.
It is sunk into the bottom of the container 10 so that it hits the right end 18 of the container 10 filled with the clarifying liquid. Next, the holding plate 19 is gently lowered into the container 10 to press the support 16 against the bottom surface 9 of the container. The holding plate may be made entirely of a transparent material, or may be made of a material that covers only the peripheral portion of the support 1 that is not related to measurement. By doing this, the first minute image center 17-1 (FIG. 1) is located at a point a certain distance a away from the right end 18 of the container 10 in the direction of the arrow 7,
From there, the image centers are aligned in the direction of arrow 7 at a constant pitch P. Next, the arm 14 is rotated counterclockwise as shown in FIG.
and start measuring.

The measurement is performed by scanning on the center line of the first minute image, that is, by scanning in the direction of arrow 8, to obtain the density pattern of the first minute image, and then moving the scanning position by pitch P in the direction of arrow 7, Scanning the center line of the image for 2 minutes is repeated n times. Scanning at this time and feeding of the scanning position are both performed by moving the mounting table 6 in the respective directions. As mentioned above, the relative position of the center line of each image with respect to the container 10 is
Since the direction of the arrow 7 is accurately determined, the positional relationship between the mounting table 6, the light receiving section 12, and the illumination section 11 at the start of scanning can be regulated so that the center of the image is scanned for the first minute. , and thereafter, by feeding at a constant feed pitch P, scanning on the image center line can be realized. Note that even if the support is slightly shifted in the direction of arrow 8, it does not matter because it has no effect on the density pattern obtained. In this example, in order to prevent the container 10 from shifting on the mounting table 6, a spring-ball mechanism 20 is provided on the left side wall of the mounting table 6 to press the container 10 against the right side wall of the mounting table 6.

The present invention is not limited to the above-mentioned example. For example, instead of moving the mounting table 6 and the container 10 to perform scanning and feeding, the light receiving section 12 and the illumination section 11 are moved to perform scanning and feeding. Good too.

Further, the scanning may be performed by moving the light receiving section 12 and the illumination section 11, and the feeding may be performed by moving the mounting table 6. In this case, the driving mechanism can be simplified. Furthermore, it is also possible to make the side surface of the clarifying liquid container transparent and place the illumination section and the light receiving section across the side surface. According to the densitometer of the present invention, since there is a support in a sufficient amount of the clarifying liquid, there is no air bubbles mixed in, and since the clarifying liquid is always inside the clearing liquid, there is little spillage to the outside, and furthermore, the transparent plate Since it does not require a separate liquid, it is easy to operate, does not stain the equipment with clarifying liquid, and allows accurate concentration measurements.

Furthermore, since one end of the illumination section or the light receiving section is immersed in the clearing liquid, diffused reflection on the surface of the clearing liquid can be removed, thereby improving density photometry accuracy.

[Brief explanation of the drawing]

Figure 1 is a diagram showing the configuration of a sample to be applied to a conventional densitometer; Figure 2 is a cross-sectional view of the sample prepared in Figure 1;
The figure is a sectional view of a sample whose upper transparent plate is made of plastic film, FIG. 4a is a longitudinal sectional view of an embodiment of the densitometer of the present invention, and FIG. 4b is a cross-sectional view of the densitometer shown in FIG. 4a. It is a diagram. 1... Support body, 6... Mounting table, 10.
......Clearing liquid solution, 11......Lighting part, 1
2... Light receiving part, 15... Clearing liquid, 1
9... Presser plate.

Claims (1)

[Claims] 1. A container with an open top for containing a clarifying solution, and a light source and a light receiving section placed oppositely on the top and bottom of this container, and the lower end of the light source or the light receiving section placed above the container is placed in the clearing solution. By changing the relative position of the clarifying liquid container and the measuring part while the support of the object to be measured is placed in the container containing the clarifying solution, the measuring part can be immersed. A densitometer for electrophoresis, characterized in that it is configured to perform scanning on each density pattern and feeding between each density pattern.