The device for electrophoresis

 

(57) Abstract:

The invention relates to medical equipment and can be used in medical institutions. The device contains a number of cells, the cell with the partition. On the wall posted holder. On the holder to be pulled in one or more membranes with cargo. The goods are situated on both sides of the holder. Goods made of magnets. Magnets zamalchivaut the ends of the membranes. The device allows to carry out the procedures necessary for diagnosis of ischemic heart disease, atherosclerosis, cancer, acute and chronic inflammatory processes in the early stages. In addition, the device improved the quality of attachment of the membranes, reduced time spent on their mount. 5 Il.

The invention relates to medical equipment and can be used in all medical institutions.

The known device for electrophoresis in the gel and membrane carriers (hereinafter membranes) patent, copyright certificates, certificates for industrial designs and Standards, issued in the name of Sobolev Century. And., patent 1780513 from 20.11.90, "Device for electrophoresis in gel", ed. St. 967470 26., GOST 27153-86 "Instruments and apparatus for medical treatment of the sample by the methods of electrophoresis."

They all contain a number of cells, each of which includes a cuvette with a partition dividing it into two cavities for a buffer that contains the holder with the membrane or gel, and cover with the electrodes. Membrane and gel are used for application of samples, and for electrical connection of the anode-cathode space through the buffer.

However, using these devices it is impossible to carry out a number of biochemical diagnostic methods, as outlined below. One of the main values is the method of attachment of the membranes.

For the implementation of new techniques required new designs with the use of membranes and holders of other structures that would simply, quickly and securely affix the membrane to cause the sample to connect the camera to a power source and to obtain a clear faction.

The prototype of the proposed device is a device according to patent 1780513 of 20 November 1990, containing a number of cells each of which includes a cuvette with a partition dividing it into two cavities for the buffer, where the holder of the gel with the work surface, with the two sides from which viparovuvanosti made strap, the upper surface of which is parallel to the working surface of the holder. On the strap overlaps at least one cellulose acetate membrane, the ends of which are entered in the supplied clamps vertical slits of the holder from its outside surface. The device is provided with a template with two parallel ribs that contact the diaphragm and the upper surfaces of the slats. The template has several Windows, the two sides each of which is made with grooves for drawing samples.

This device is convenient for mounting single bands membranes width of 28 mm in the study of a small number of samples, but inconvenient when attaching membranes standard size 90 x 90 mm 180 x 90 mm, Therefore, the disadvantage of this device is that the time required to mount six lanes membranes width 28 mm, is 15 minutes In the inventive device, the time for fixing membrane widths from 28 to 180 mm is 0.5 minutes

The aim of the invention is the possibility of the following methods:

- electrophoretic fractionation of proteins and lipoproteins serum;

- electrophoretic separation of iovernment lactate dehydrogenase and melachroinos besieged-lipoproteidna faction.

All these techniques are tested on the claimed device at the Department of biochemistry Petersburg sanitary Academy.

The proposed device and methods necessary for the diagnosis of lipid metabolism that occur in ischemic heart disease and atherosclerosis, cancer, acute and chronic inflammatory processes in the early stages.

The proposed device is necessary when determining the purity in the manufacture and use-globulin preparations, for example, blood transfusion stations.

This objective is achieved in that the holder is stretched one or more membranes with the help of goods that are located on both sides of the holder, for example, magnets, locking the ends of the membranes.

The proposed device for electrophoresis, (hereinafter "device") contains one or more electrophoresis chambers and power supply (not shown).

In Fig.1 shows a camera for electrophoresis; Fig.2 and 3 the holder of Fig.4 and 5 the template.

Camera (Fig.1) contains a cuvette with 1 partition 2 dividing it into two cavities for buffer 3, which is hosting the holder 4 with the membrane athisaya membrane.

The device operates as follows.

1. The device uses membrane "VLADIPOR" MFA-MA 4 or 5, which has a standard size of 90 x 90 mm 180 x 90 mm TU 6-05-1903-87. The device allows you to secure the membrane of any width, standard or cut into strips, almost instantly.

2. Uses a buffer of the following composition:

- barbitala of sodium was 12.75 g;

- barbitala - 2.3 g - fill with distilled water to 1000 ml and placed in a water bath.

3. Prepare a 7% solution of acetic acid, to which 7 ml of glacial acetic acid diluted with distilled water to 100 ml.

4. For staining of proteins prepared 0.25% solution amidating dye, for which 250 mg of dye dissolved in 100 ml of 7% acetic acid.

5. For staining of lipoproteins using Sudan black, Sudan 3 or Sudan 4 (red). A small amount of dye on the tip of a scalpel placed in 5 ml of isopropyl alcohol. After the dye is completely dissolved, add 10 ml of 5% Paon.

6. For enlightenment membranes use solution consisting of methanol - 87 ml, glacial acetic acid (12 ml) and glycerol - 1 ml of This solution is suitable only when the fractionation of proteins.

Paradol 4 (cuvette, holder, template and cover have locks 9, which prevent incorrect installation.)

2. On the working surface of the dry membrane ballpoint pen put the arrow indicating the direction of fractionation. Arrow is applied across the grain. The working surface has a porous structure, and non - smooth.

3. The membrane is placed for 1-3 min in a buffer solution, is poured into a separate container.

4. The membrane is removed from the buffer and put between two sheets of filter paper side up.

5. Remove top sheet of filter paper. Zamalchivaut magnets each of those ends of the membrane, which is perpendicular to the arrow.

6. The membrane 5 is placed on the holder 4, as shown in Fig.1, 2 and 3.

7. On the cuvette 1 set the template 8.

8. Applicator (not shown) through the slot 10 is applied to the sample (serum). The template is removed.

9. The cuvette is closed by a cover 11 with the electrodes 12.

10. The camera is supplied a constant regulated voltage whose magnitude is determined by the method.

11. After electrophoresis, the membrane is placed in a fixing solution for 10 min, then in okrashivaemuyu device

1. The main advantage of this device is the implementation of the above diagnostic methods.

2. Significantly reduced time to mount the membranes of any width, superior quality mounting membranes.

3. Membranes are continuously metered tension, which is very important, because when conducting electrophoresis membranes become wet, stretch and SAG, which affects the process of electrophoresis.

4. When applying the sample applicator membrane soft springs, which eliminates the gap between her and returns to its original position.

Process automation

1. The device connects to the power source by a simple camera moves into the connector. Voltage is applied to one turn of the handle timer this sets the time of electrophoresis. The camera turns off automatically.

2. Among the available processing methods electrophoregram preferred method of densitometrically. However, in the last time our computer implemented method of processing electrophoregram.

At the present time on the basis of S-Petersburg scientific research Institute of cardiology of Russian Federation Ministry of health we have developed a computer system for processing electrophorese tool for the automated processing of electrophoregram.

The system allows to automatically determine the percentage of fractions and mass concentration in g/l, to detect the presence of abnormal proteins "paraproteins", not typical of the blood of a healthy person, to detect the absence of factions, to capture changes in the structure of any protein, to approximate electrophoregram depending on the input to the program corrections, etc.

3. The device will be included in the set of devices that are commercially available and have been used successfully for over fifteen years:

the electrophoretic device PAPAG-1 TU 64-1-3174-80, registration card 78/626-82,

devices immunoelectrophoretic PIAF-01-, PEEP-01-1, PIF-01-2, PIF-01-3 TO-1-3945-85, reg. ID 96/386-178. These instruments are included in the State REGISTER of medical products of the Ministry of Health and medical industry of the Russian Federation, M., 1996, page 124.

The device for electrophoresis containing a number of chambers, each of which includes a cuvette with a partition dividing it into two cavities for the buffer, which holds the holder with cellulose acetate membrane cap with electrodes and a template, wherein the holder tight one and the

 

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