Method of staining blood smears for microscopic determination of structural organisation of cell phase activity

G01N1/30 - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (separating components of materials in general B01D, B01J, B03, B07; apparatus fully provided for in a single other subclass, see the relevant subclass, e.g. B01L; measuring or testing processes other than immunoassay, involving enzymes or micro-organisms C12M, C12Q; investigation of foundation soil in situE02D0001000000; monitoring or diagnostic devices for exhaust-gas treatment apparatus F01N0011000000; sensing humidity changes for compensating measurements of other variables or for compensating readings of instruments for variations in humidity, seeG01D; or the relevant subclass for the variable measuredtesting or determining the properties of structures G01M; measuring or investigating electric or magnetic properties of materials G01R; systems in general for determining distance, velocity or presence by use of propagation effects, e.g. Doppler effect, propagation time, of reflected or reradiated radio waves, analogous arrangements using other waves G01S; determining sensitivity, graininess, or density of photographic materials G03C0005020000; testing component parts of nuclear reactors G21C0017000000)

FIELD: medicine.

SUBSTANCE: method includes the preparation of smear from peripheral blood with preliminary fixation with methyl alcohol, drying, washing with distilled water. After that, the smears are placed in a potassium chloride solution in a ratio of 0.57 g of potassium chloride per 100 ml of distilled water for 20 min and washed with distilled water. Additionally prepared is a mixture of solutions, prepared ex tempore, containing a solution "A" and "B". The solution "A" includes a 50% silver nitrate solution in an amount of 5 g of silver nitrate + 5 ml of distilled water. The solution "B" includes a 2% solution of gelatin on a 1% formic acid solution in an amount of 15.8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml of 10% gelatin. The solutions "A" and "B" are mixed in an amount of 5 ml of each, in darkness, with further submergence of the blood smears for 20 min in darkness in the thermostat at a temperature of 37°C with the further submergence of the smears into distilled water for 2-3 seconds. After that, they are twice subjected to a 8 min exposure in a 5% sodium thiosulphate solution in darkness in the thermostat at a temperature of 37°C. After that, they are washed successively with tap water and distilled water, after-staining is performed in the Romanovskiy dye for 30 min. After that, the smears are washed again with tap water, air-dried, placed in the Canadian balm and covered with a coverslip.

EFFECT: increased quality of smear staining and provision of a possibility to identify and further evaluate parameters of nucleolus organiser regions.

4 tbl, 6 ex

 

The technical field to which the invention relates

The invention relates to veterinary science and medicine, in particular to a method for staining of blood smears for microscopic determination of structural organization and phases of cell activity, and can be used in cytophysiology, cytopathology and cytogenetics, and also recommended for use in the haematological studies in determining the functional state of blood cells.

The level of technology

A method of staining blood smears, which is that for the preparation of blood smears them fix solution Nikiforov, the painting is carried out in two dyes, which take the Ehrlich's hematoxylin and eosin sodium, at first dabs placed in aged in sealed containers within 360 days, a solution of Ehrlich's hematoxylin for 2-3 minutes, then submerge them in a container of tap water for 1-2 minutes until you're blue smears, then the strokes are dried with filter paper and placed in 1% aqueous eosin solution-sodium for 1-2 minutes, washed stained smears with distilled water for 8-10 C and dried in air (Patent RU №2304776 "Method of staining blood smears" / V. I. Trukhachev, V. Rodin, V. V. Mikhailenko, A. A. Dergunov / IPC G01N 33/48, G01N 1/30, publ. 20.08.2007).

The disadvantages of this method are:

- lack of opportunities� identify areas of nucleolar organizers in connection with the non-specific dye;

- higher complexity and economic cost of the method.

A method of staining blood smears, lies in the fact that smears fixed and stained with lowering them 4-5 times within one second in solutions of anionic dye - 0,5% solution of eosin, and then a cationic dye - 0,5% solution of Azura, and separately in 0.5% solution of methylene blue. The method allows to speed up the painting strokes and improve the quality of smears for microscopic examination (Patent RU №2081404, "the Method of staining blood smears" / S. A. Pozov, I. I. Kolomytsev / IPC G01N 1/30, G01N 1/28, publ. 10.06.1997).

The disadvantage of this method is:

the inability identify areas of nucleolar organizers, which does not allow to judge about the functional activity of cells.

A method of staining blood smears, lies in the fact that blood smears are thermally fixed at 160-180°C, stained with 0.25% aqueous solution of eosin for 110-130 C, rinsed with water, and then stained with a 0.2% aqueous solution diazepamillegalgu benzthiazole with 50-70 (Patent RU №2044295 "Method of staining blood smears" / V. V. Rogovin, S. I. Chikalov, I. G. Han, S. P. Titov // IPC G01N 1/30, publ. 20.09.1995).

The disadvantage of this method is the inability to identify areas of nucleolar organizers in connection with nespecificnomu dye.

The most� closest to the technical essence and the achieved positive effect and adopted by the authors for the prototype is a method of staining by the method of Howell and black with little modifications, aimed at identifying areas of nucleolar organizers. The cells are fixed with cold mixture of methanol and glacial acetic acid (3:1) for 30 min. the cells are washed sequentially in 3 shifts): 50%, 30% ethanol with a flow of distilled water, dried in air. For staining preparations prepare two solutions: the first consists of 2% gelatin and 1% (w/w) formic acid in bidistilled water; the second is 50% AgNO3bidistilled water. On a clean glass slide, put a drop of gelatin solution and two drops of silver nitrate solution. On the drop put the coverslip with cells facing down. The painting is carried out in a humid chamber in a thermostat at 60°C. staining Time varies from 4 to 11 min. control over the intensity of staining is carried out visually using an optical microscope MBI-3 (lens ×10, eyepiece ×10). Painted glass washed with distilled water until the water until it becomes transparent. Then, in order to avoid discoloration of drugs over time, the glass cell is placed at 30 C in a 5% solution of sodium hyposulphite, and then the glass is washed with water, dehydrated in alcohols and conclude in canadian balsam (see method of staining by the method of Howell and black, Howell M., Black D. A., 1980. Controlled silverstaining of nucleolus organiser regions with a protective colloidal developer. A 1-step method. Experietia. V. 36, P. 1014-1015).

This method of painting has the following disadvantages:

- higher complexity operations when staining;

- has a long painting process;

- no possibility of morphological identification of cells.

Disclosure of the invention

The object of the invention is to provide a method of staining blood smears for microscopic determination of structural organization and phases of cell activity with the reduction of the time of receipt of large and small batches efficiently stained blood smears, in which it is possible to study areas of nucleolar organizers to determine the functional activity of blood cells (see tab.1-4).

The technical result that can be achieved using the present invention is to reduce time and improve the quality of staining of smears, with the ability to identify and then estimate the parameters of the zones of nucleolar organizers.

The technical result is achieved by using the method of staining blood smears for microscopic determination of structural organization and phase activity of cells, comprising preparing a smear from peripheral blood pre-commit methyl alcohol, drying, washing with distilled water, followed by placing in a solution of KCl at a ratio of 0.57 g KCl n� 100 ml of distilled H 2O for 20 min, and washing with distilled water, optionally a mixture of solutions prepared ex tempore containing solution "A" containing 50% solution of silver nitrate, in an amount of 5 g of AgNO3+ 5 ml of distilled water, and solution "B", including a 2% solution of gelatin in 1% formic acid in the amount of 15.8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml of 10% gelatin, then mix the solutions "A" and "b" in the amount of 5 ml each, in the dark, and to this mixture, immerse the slides for 20 min in the dark in a thermostat at a temperature of 37°C, followed by immersion of the stroke for 2-3 seconds in distilled water, then incubated twice for 8 min in a 5% solution of Na thiosulfate in the dark in a thermostat at a temperature of 37°C, washed successively with tap water and distilled water, conduct bookrack in the paint Romanowski for 30 min, washed again with tap water, dried smears on the air, sign in canadian balsam and cover with cover glass.

Thus, the proposed method allows for the haematological studies in medicine and veterinary medicine to conduct rapid analysis of changes in the structure and phases of the functional activity of blood cells, to give opinions about the intensity and stability of the physiological PR�processes, to identify the features that determine the prognosis of the disease.

The inventive method of staining blood smears for microscopic determination of structural organization and phase activity of cells is as follows. Prepared smears from peripheral blood pre-fixed with methyl alcohol, dried under suction and washed with distilled water. Placed in a KCl solution (0,57 g of KCl in 100 ml distilled H2(O) for 20 min and Then washed with distilled water. Placed in a mixture of solutions prepared ex tempore. Solution "A" - 50% solution of silver nitrate (5 g of AgNO3+ 5 ml distilled water). Solution "B" - 2% solution of gelatin for laboratory works on a 1% solution of formic acid (15,8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml 10% gelatin for laboratory work). Mix a solution A (5 ml) and b (5 ml) in the dark, and to this mixture, immerse the slides for 20 min in the dark in a thermostat at a temperature of 37°C. After that swabs dipped for 2-3 seconds in distilled water, then incubated twice for 8 min in a 5% solution of Na thiosulfate in the dark in a thermostat at a temperature of 37°C. Washed with tap water, then distilled water. Spend bookrack paint Romanowski for 30 min. Washed with tap water and dried smears on the air. Then zakluchautsya in canadian balsam and cover with cover glass. Spend microscopy of smears to determine and estimate the parameters of the zones of nucleolar organizers.

Brief description of the drawings and other materials

Table 1 provides the method of staining blood smears for microscopic determination of structural organization and phases of cell activity, the number OAOR in the nuclei of red blood cells in geese of different ages, pieces.

Table 2 - the same size OAOR in the nuclei of erythrocytes geese of different ages, mcm-square

In table 3, the number OAOR in the nuclei of erythrocytes in ducks of different ages, pieces.

Table 4 - the same size OAOR in the nuclei of erythrocytes ducks of different ages, mcm-square

The implementation of the invention

Examples of specific implementation method of staining blood smears for microscopic determination of structural organization and phase activity of cells

Example 1. With the aim of staining taken party in the amount of 30 strokes. Coloring is as follows. Prepared smears from peripheral blood pre-fixed with methyl alcohol, dried under suction and washed with distilled water. Placed in a KCl solution (0,57 g of KCl in 100 ml distilled H2(O) for 20 min and Then washed with distilled water. Placed in a mixture of solutions prepared ex tempore. Solution "A" - 50% solution of silver nitrate (5 g of AgNO3+ 5 ml distilled water). Rast�or "In" - 2% solution of gelatin for laboratory works on a 1% solution of formic acid (15,8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml 10% gelatin for laboratory work). Mix a solution A (5 ml) and b (5 ml) in the dark, and to this mixture, immerse the slides for 15 min in the dark in a thermostat at a temperature of 30°C. After that swabs dipped for 2-3 seconds in distilled water, then incubated twice for 5 min in a 5% solution of Na thiosulfate in the dark in a thermostat at a temperature of 37°C. Washed with tap water, then distilled water. Dried smears on the air. Then make strokes in canadian balsam and cover with cover glass. Spend microscopy of smears to determine and estimate the parameters of the zones of nucleolar organizers.

In analysing the data, noted the following:

areas of nucleolar organizers are poorly visualized, and were fuzzy;

- define the settings of the zones of nucleolar organizers was complicated;

- over time, the quality of smear deteriorated, worsened the visualization of the areas.

Example 2. With the aim of staining taken party in the amount of 30 strokes. Coloring is as follows. Prepared smears from peripheral blood pre-fixed with methyl alcohol, dried under suction and washed with distilled water. Pomeshaut KCl solution (0,57 g of KCl in 100 ml distilled H 2(O) for 20 min and Then washed with distilled water. Placed in a mixture of solutions prepared ex tempore. Solution "A" - 50% solution of silver nitrate (5 g of AgNO3+ 5 ml distilled water). Solution "B" - 2% solution of gelatin for laboratory works on a 1% solution of formic acid (15,8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml 10% gelatin for laboratory work). Mix a solution A (5 ml) and b (5 ml) in the dark, and to this mixture, immerse the slides for 20 min in the dark in a thermostat at a temperature of 37°C. After that swabs dipped for 2-3 seconds in distilled water, then incubated twice for 8 min in a 5% solution of Na thiosulfate in the dark in a thermostat at a temperature of 37°C. Washed with tap water, then distilled water. Dried smears on the air. Then sign in canadian balsam and cover with cover glass. Spend microscopy of smears to determine and estimate the parameters of the zones of nucleolar organizers in tables 1-4 presents examples of determination OAOR in the nuclei of red blood cells in geese of different ages, NJ; area OAOR in the nuclei of erythrocytes geese of different ages, microns square; number OAOR in the nuclei of erythrocytes in ducks of different ages, NJ; area OAOR in the nuclei of erythrocytes ducks of different ages, mcm-square

In analysing the data, noted the following�:

areas of nucleolar organizers Visualizers clearly;

- define the settings of the zones of nucleolar organizers available in a light microscope;

- the storage time of the stroke did not affect the visual quality zones.

Example 3. With the aim of staining taken party in the amount of 30 strokes. Coloring is as follows. Prepared smears from peripheral blood pre-fixed with methyl alcohol, dried under suction and washed with distilled water. Hurt in a KCl solution (0,57 g of KCl in 100 ml distilled H2(O) for 20 min and Then washed with distilled water. Hurt in a mixture of solutions prepared ex tempore. Solution "A" - 50% solution of silver nitrate (5 g of AgNO3+ 5 ml distilled water). Solution "B" - 2% solution of gelatin for laboratory works on a 1% solution of formic acid (15,8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml 10% gelatin for laboratory work). Mix a solution A (5 ml) and b (5 ml) in the dark, and to this mixture, immerse the slides for 25 min in the dark in a thermostat at a temperature of 60°C. After that swabs dipped for 2-3 seconds in distilled water, then incubated twice for 10 min in a 5% solution of Na thiosulfate in the dark in a thermostat at a temperature of 37°C. Washed with tap water, then distilled water. Dried smears on �ozdok. Then sign in canadian balsam and cover with cover glass. Spend microscopy of smears to determine and estimate the parameters of the zones of nucleolar organizers.

In analysing the data, noted the following:

- repainting of the areas of nucleolar organizers;

- it is noted excessive darkening of the image, which complicates the study areas of nucleolar organizers.

Example 4. With the aim of staining taken party in the amount of 30 strokes. Coloring is as follows. Prepared smears from peripheral blood pre-fixed with methyl alcohol, dried under suction and washed with distilled water. Placed in a KCl solution (0,57 g of KCl in 100 ml distilled H2(O) for 20 min and Then washed with distilled water. Hurt in a mixture of solutions prepared ex tempore. Solution "A" - 50% solution of silver nitrate (5 g of AgNO3+ 5 ml distilled water). Solution "B" - 2% solution of gelatin for laboratory works on a 1% solution of formic acid (15,8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml 10% gelatin for laboratory work). Mix a solution A (5 ml) and b (5 ml) in the dark, and to this mixture, immerse the slides for 20 min in the dark in a thermostat at a temperature of 37°C. After that swabs dipped for 2-3 seconds in distilled water, then incubated� twice for 8 min in a 5% solution of Na thiosulfate, in the dark in a thermostat at a temperature of 37°C. Washed with tap water, then distilled water. Spend bookrack safranin T for 24 h. Washed with tap water and dried smears on the air. Carry out the dehydration of alcohols. Then make strokes in canadian balsam and cover with cover glass. Spend microscopy of smears to determine and estimate the parameters of the zones of nucleolar organizers.

The analysis of the data showed that in the entire group of strokes by 80% occurred leaching paint alcohols, whereby to determine the nucleolar organizers and conduct morphological identification of blood cells was not possible.

Example 5. With the aim of staining taken party in the amount of 30 strokes. Coloring is as follows. Prepared smears from peripheral blood pre-fixed with methyl alcohol, dried under suction and washed with distilled water. Placed in a KCl solution (0,57 g of KCl in 100 ml distilled H2(O) for 20 min and Then washed with distilled water. Hurt in a mixture of solutions prepared ex tempore. Solution "A" - 50% solution of silver nitrate (5 g of AgNO3+ 5 ml distilled water). Solution "B" - 2% solution of gelatin for laboratory works on a 1% solution of formic acid (15,8 ml of distilled water + 0.2 ml 100% mu�avinou acid + 4.0 ml 10% gelatin for laboratory work). Mix a solution A (5 ml) and b (5 ml) in the dark, and to this mixture, immerse the slides for 20 min in the dark in a thermostat at a temperature of 37°C. After that swabs dipped for 2-3 seconds in distilled water, then incubated twice for 8 min in a 5% solution of Na thiosulfate in the dark in a thermostat at a temperature of 37°C. Washed in tap water, then distilled water. Spend bookrack green diamond for 2 min, Washed with tap water and dried smears on the air. Then make strokes in canadian balsam and cover with cover glass. Spend microscopy of smears to determine and estimate the parameters of the zones of nucleolar organizers.

The analysis of the obtained data revealed that when decresce green diamond all cellular structures acquired an intensely green color. Consequently, the study of parameters of nucleolar organizers seemed not possible.

Example 6. With the aim of staining taken party in the amount of 30 strokes. Coloring is as follows. Prepared smears from peripheral blood pre-fixed with methyl alcohol, dried under suction and washed with distilled water. Placed in a KCl solution (0,57 g of KCl in 100 ml distilled H2(O) for 20 min and Then washed with distilled water. Placed in a mixture of solutions, �prigotovlennyh ex tempore. Solution "A" - 50% solution of silver nitrate (5 g of AgNO3+ 5 ml distilled water). Solution "B" - 2% solution of gelatin for laboratory works on a 1% solution of formic acid (15,8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml 10% gelatin for laboratory work). Mix a solution A (5 ml) and b (5 ml) in the dark, and to this mixture, immerse the slides for 20 min in the dark in a thermostat at a temperature of 37°C. After that swabs dipped for 2-3 seconds in distilled water, then incubated twice for 8 min in a 5% solution of Na thiosulfate in the dark in a thermostat at a temperature of 37°C. Washed with tap water, then distilled water. Spend bookrack paint Romanowski for 30 min. Washed with tap water and dried smears on the air. Then make strokes in canadian balsam and cover with cover glass. Spend microscopy of smears to determine and estimate the parameters of the zones of nucleolar organizers.

The analysis of the data led to the conclusion that 85% of strokes successfully painted and they can study how the structure of cells and nucleolar organizers. The proposed method of coloring is superior to the most commonly used method of staining for the detection of nucleolar organizers by Howell and black (see Howell, Black, 1980) that along with the study of the nucleolus�'s organizers got the opportunity to explore the rest of the cell structure, the method significantly reduces the time associated with the color.

Thus, based on the above examples revealed that the optimal are the examples 2 and 6, by which it is possible to study areas of nucleolar organizers to determine the functional activity of blood cells, while in tables 1-4 presents examples of determination OAOR in the nuclei of red blood cells in geese of different ages, NJ; area OAOR in the nuclei of erythrocytes geese of different ages, microns square; number OAOR in the nuclei of erythrocytes in ducks of different ages, NJ; area OAOR in the nuclei of erythrocytes ducks of different ages, mcm-square

The present invention compared with the prototype and other known technical solutions have the following advantages:

- reduced time staining;

- the method allows to obtain swabs high quality staining;

- the method allows to visualize the nucleolar organizers along with all other structures of the cell.

Table 1
The number OAOR in the nuclei of red blood cells in geese of different ages, EA.
The stage of the experimentThe experimental group
males, M±m (n=300)females, M±m (n=300)
1 day6,22±0,128,23±0,14##
1 month10,02±0,12**Of 9.55±0,11**##
3 months7,90±0,09**7,68±0,08**
6 monthsOf 11.45±0,11**11,52±0,10**
9 months9,92±0,11**8,92±0,09**##
12 months9,00±0,08**9,10±0,09
Note: statistical significance of the differences with the earlier period: * - p<0,05, ** - p<0.01; with males of the same period: # - p < 0,05, # # p<0.01.

Table 2
Size OAOR in the nuclei of erythrocytes geese of different ages, mcm sq
The stage of the experimentThe experimental group
males, M±m (n=300)females, �±m (n=300)
1 day0,293±0,0090,404±0,007##
1 monthOf 0.382±0,009**0,425±0,011##
3 months0,368±0,0070,356±0,010**
6 months0,297±0,007**0,262±0,008**##
9 months0,343±0,012**0,337±0,011**
12 months0,234±0,008**0,244±0,009**
Note: statistical significance of the differences with the earlier period: * - p<0,05, ** - p<0.01; with males of the same period: # - p < 0,05, # # p<0.01.

Table 3
The number OAOR in the nuclei of erythrocytes in ducks of different ages, EA.
The stage of the experimentThe experimental group
females, M±m (n=300)males, M±m (n=300)
1 day 5,12±0,146,14±0,07##
1 month7,86±0,08**8,75±0,09**##
3 months6,37±0,12**6,91±0,11**##
6 months10,80±0,07**11,74±0,09**##
9 months11,14±0,09**10,43±0,11**##
12 months11,06±0,1011,59±0,13**##
Note: statistical significance of the differences with the earlier period: * - p<0,05, ** - p<0.01; with females of the same period: # - p < 0,05, # # p<0.01.

Table 4
Size OAOR in the nuclei of erythrocytes ducks of different ages, mcm sq
The stage of the experimentThe experimental group
females, M±m (n=300)males, M±m (n*300)
1 day0,384±0,008 0,337±0,007##
1 month0,664±0,009**0,372±0,009**##
3 months0,469±0,008**0,527±0,011**##
6 months0,431±0,010**0,391±0,008**##
9 months0,435±0,0110,414±0,010
12 months0,378±0,007**0,330±0,008**##
Note: statistical significance of the differences with the earlier period: * - p<0,05, ** - p<0.01; with females of the same period: # - p < 0,05, # # p<0.01.

The method of staining blood smears for microscopic determination of structural organization and phase activity of cells, comprising preparing a smear from peripheral blood pre-commit methyl alcohol, drying, washing with distilled water, followed by placing in a solution of potassium chloride in a ratio of 0.57 g KCl in 100 ml distilled water for 20 min, and washing with distilled water, characterized in that it further prepare the mixture of solutions prepared ex tempore containing solution "A", include�rd 50% solution of silver nitrate, in an amount of 5 g of silver nitrate + 5 ml of distilled water, and solution "B", including a 2% solution of gelatin in 1% formic acid in an amount of 15.8 ml of distilled water + 0.2 ml of 100% formic acid + 4.0 ml of 10% gelatin, then mix the solutions "A" and "b" in the amount of 5 ml each in the dark, and to this mixture, immerse the slides for 20 min in the dark in a thermostat at 37°C followed by immersion of the stroke for 2-3 seconds in distilled water, then incubated twice for 8 min in 5% sodium thiosulfate solution in the dark in a thermostat at temperature 37°C, washed successively with tap water and distilled water, conduct bookrack in the paint Romanowski for 30 min, washed again with tap water, dried smears on the air, sign in canadian balsam and cover with cover glass.



 

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9 cl, 9 dwg

FIELD: medicine.

SUBSTANCE: invention deals with a method of evaluating a threat of hypoxia formation in pregnant women with the aggravation of the cytomegaloviral infection in the third trimester of gestation. The essence of the method: in peripheral blood a titre of antibodies to cytomegalovirus is determined, the activity of phosphoenolpyruvate dehydrogenase is determined by the histochemical method, the content of discocytes, echinocytes, target-shaped erythrocytes, degenerative forms of erythrocytes, oxyhaemoglobine quantity are determined in peripheral blood. If the titre of antibodies to cytomegalovirus is 1:1600, detection of the reduction of phosphoenolpyruvate dehydrogenase activity to 12.0±0.09 c. u.: reduction of discocytes in peripheral blood to 74.0±1.3%, increase of the number of echinocytes to 7.0-0.2, target-shaped erythrocytes to 8.5±0.3%, degenerative forms of erythrocytes to 10.5±0.15% and reduction of the oxyhaemoglobine quantity to 92.0±1.6% a threat of formation of haemic hypoxia is evaluated.

EFFECT: improvement of the method of the threat evaluation.

2 dwg

FIELD: medicine.

SUBSTANCE: test tray comprises a case 1 made of an optically transparent material. From one end face, the case 1 has a stop plug 2 with a hole 3, a connecting pipe 4 threaded to connect to either a haemofilter 5, or a cap 7. The case 1 comprises a movable piston 8 connected by a rod 10 to a handle 11. Electrically supplied electrodes 12 are arranged on surfaces of the stop plug 2, piston 8 and on the inner surface of the case 1. The electrodes 12 are connected to contact groups of a device - a laser analyser - through conductors 13, 14. What is disclosed is an alternative version of the structural embodiment of the test tray.

EFFECT: sterile measurement of the colloidal fluid sample.

17 cl, 5 dwg

FIELD: agriculture.

SUBSTANCE: method of selection of horizontal soil monolith comprises embedding along the genetic horizons of nth thin-walled metal cylinder-monolith-selector of the ith diameter with a pointed lower end of a triangular shape. The selection of the horizontal soil monolith from the pit is carried out with the number of cylinders-monolith-selectors k, equal to where i - number of the cylinder diameter (n > i > 1), n - number of cylinders of different diameters, ki - the number of repetitions of the cylinder of ith diameter (ki > 3). And each time, prior to selection of the horizontal soil monolith the inner surface of each used cylinder-monolith-selector is greased with petroleum jelly, and the load on the cylinder-monolith-selector is performed in a direction perpendicular to the surface of the pit stepwise with fixing the load of each step. The set of devices for selection of the horizontal soil monolith comprises the said k-th number of thin-walled metal cylinder-monolith-selectors and a metal cylindrical nozzle on the cylinder-monolith-selector. The metal nozzle is provided with a cylindrical recess in one of its ends, which diameter is equal to the outer diameter of the cylinder-monolith-selector having a maximum diameter of n cylinder-monolith-selectors, and the axis of symmetry coincident with the axis of symmetry of the metal cylindrical nozzle. The set also comprises (n-1) washer with an outer diameter equal to the diameter of the recess and the thickness equal to the height of the recess in the end of the metal cylindrical nozzle with the ability of mounting of each of them to the recess, followed by fixing in it. The inner washer diameters are different and equal to the outer diameter of each of the (n-1) cylinder-monolith-selector, constituting a pair: washer-cylinder-monolith-selector. Set is provided with a screw press with a head and a heel of cylindrical shape and a shield with a recess on its axis of symmetry with the ability of mounting in it through the heel of the screw press. And in the other end of the metal cylindrical nozzle on the cylinder-monolith-selector on its axis of symmetry a recess of cylindrical shape is made, the diameter and depth of which correspond to the diameter and thickness of the head of the screw press.

EFFECT: improvement of quality of sampling soil of undisturbed placement and increase in the accuracy of determining the water-physical and filtration properties of soil on genetic horizons of the soil profile, reduction of time for selection of the monolith and the complexity of operations in selection of quality horizontal soil sample.

3 cl, 1 dwg, 1 tbl

FIELD: engines and pumps.

SUBSTANCE: device comprises a floating element 10, which is placed onto the sea surface and connected to a pump, rigidly fixed to the sea bottom or to massive floatage 8. The pump is made in the form of a cylindrical pipe-shaped vertically arranged chamber 1 semi-submerged into the sea, which in its upper and lower parts is equipped accordingly with lower 3 and upper 6 nozzles. At the lower nozzle 3 there is a hose 4 with certain length arranged in water depth. In the chamber there is a piston in the form of an inlet check valve placed on the stem 9, which is made as capable of passing water in the chamber only in direction from the lower nozzle to the upper one and is connected by means of the stem 9 with a floating element 10. The piston may be made within a membrane 12 adjacent to the plane of a disc 11 made with through holes, axes of which are parallel to the axis of the disc.

EFFECT: simplified design, expanded area of application of a device for water lifting.

2 cl, 1 dwg

FIELD: measurement equipment.

SUBSTANCE: invention relates to equipment for determining consumptions and periodic water sampling from different horizons of a peat deposit, which are fixed as to depth. The complex includes a well casing pipe with a cone tip and a water intake structure. Besides, a sampling unit includes a cylindrical housing, on which there located are two elastic rubber cuffs with diameter equal to well diameter; in the wall of the cylindrical housing there are side holes - a middle one - for water receiving from a working horizon and is located between two cuffs; an upper one is located above the upper cuff; the lower one is located under the lower cuff; upper and lower holes are of a transit type and connected to each other with a tube passing inside the cylindrical housing of the sampling unit; the lower part of the cylindrical housing is connected to the water intake structure through a flange attached to the cylindrical housing; the upper part of the cylindrical housing is connected to a bracket for lifting the sampling unit and the water intake structure connected to it, the diameter of which is lower than inner diameter of the well casing pipe; the well casing pipe is pipes from one to N, which are connected to each other with external threaded couplings and side holes made throughout length of the pipes.

EFFECT: simpler design.

2 cl, 2 dwg

FIELD: measurement equipment.

SUBSTANCE: invention relates to a measuring probe for measurement and taking samples in molten metal. The probe is provided with a measurement head located on a rod, which includes at least a temperature sensor and a sampling chamber. The latter is at least partially enveloped with the measurement head and includes an input channel passing through the measurement head. The input channel has an internal section with length L, which is located in the measurement head, and has minimum diameter D at least at one point in this internal section; with that, L/D2 ratio is less than 0.6 mm-1. Besides, the measurement head has counter pressure Pg of lower than 20 mbar, which is determined so that first a reference gas flow is passed via a pipe with two open ends, and pressure P1 is measured in the pipe. Then, the pipe is inserted with one end into the inlet channel of the measurement head; the same reference gas flow is passed via the pipe and pressure P2 is measured in the pipe, and counter pressure Pg of the measurement head is determined based on difference P2-P1.

EFFECT: improvement of quality of the obtained samples.

23 cl, 5 dwg

FIELD: chemistry.

SUBSTANCE: apparatus includes a receptacle in the form of a sealed container whose lower part is fitted with a controlled two-way diaphragm valve, and a nozzle for feeding water used to wash the sample delivery line. The apparatus also includes a filter element, which is hermetically connected to a filtrate storage container, and an information control, display and transmission unit. The filter element is located in the working medium and is mounted on the sample removal line. To regenerate the filter surface, the apparatus includes a hydraulic pressure pulsator, which is installed on the sample removal line between a ball valve and a resistance disc, through which the sample is fed into the sample receptacle in the form of a sealed container, having a water-cooled jacket located at the end of the filtrate removal line.

EFFECT: invention improves the accuracy of monitoring process parameters, provides timely detection and rectification of process disorders, which enable to obtain more reliable data on the chemical composition of a solution.

1 dwg

FIELD: medicine.

SUBSTANCE: blood is sampled, acidified to pH 2-3 with aqueous oxalic acid, extracted in toluene for 5 min; the prepared extract is centrifuged for 60 min at 7,000 rpm, added with sodium sulphate to dewater and acetylated for 3 hours by introducing trifluoroacetic anhydride while stirring continuously in the pyridine medium. The blood sample, toluene, trifluoroacetic anhydride and pyridine are taken in volume ratio 5:2.5:0.2:0.1 respectively.

EFFECT: simplifying the stage of sample preparation and increasing the sensitivity of pentachlorophenol test.

3 cl, 4 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: described are: solution for preliminary processing for immunohistochemical staining, which elutes paraffin-containing mounting medium from microscope slide with tissue sample, embedded into medium, and extracts antigenicity of tissue sample, and which can be used three or more times, and solution concentrate for preliminary processing for immunohistochemical staining, which provides possibility of easy obtaining of solution for preliminary processing. Solution for preliminary processing for immunohistochemical staining contains antigen-extracting agent, certain non-ionic surface-active substances in specified range and cyclodextrin or its derivative in certain amount, with not less than 80% of water constituting the remaining part. Composition of antigen-extracting agent is such that pH of solution for preliminary processing is in specified range, and content of cyclodextrin or its derivative represents specified amount.

EFFECT: obtaining solutions for preliminary processing for immunohistochemical staining.

21 cl, 5 tbl, 11 ex

FIELD: test equipment.

SUBSTANCE: invention relates to laboratory test equipment, namely to the device for forming and testing of samples of thin coatings in loading devices, for example, for testing of thin ceramic heat-shielding coatings for tensile strength. The device is a two-piece unit intended for placement in the load device comprising two cylindrical and circular details the external surface of which is intended for application, at least, of one layer of thin coating and forming of a sample. One of cylindrical details has on the axis a cylindrical cavity, and another one a companion cylindrical ledge placed through a ring hole in a cavity and connecting the details. The external surface of cylindrical details has adhesion, and a ring surface has applied coatings without adhesion, and serve, respectively, for forming of a sample as a connecting layer and/or non- adhesion thin coating.

EFFECT: improvement of reliability of study of strength properties of thin coatings by forming of non-adhesion longitudinal superficial sample on the two-piece unit suitable for loading by longitudinal and temperature loads.

5 cl, 3 dwg

FIELD: chemistry.

SUBSTANCE: group of inventions relates to making preparations of adhering or non-adhering cells and/or particles, contained in liquid. Compartment (10) for making said preparations contains accumulation chamber (20) for storing liquid in accumulation chamber in suspended state against force of gravity, acting on liquid, only due to forces of adhesion and/or superficial tension. Accumulation chamber is made with possibility of storing liquid, which contains cells and/or particles, and discharge of stored liquid, containing said cells and/or particles, through output opening (22) by application of specified external force, in particular centrifugal force. Compartment contains channel (30), located adjacently to output opening (22) of accumulation chamber (20), with output opening (22) of accumulation chamber (20) leading to said channel. Channel (30) has section, larger than section of output opening (22), and wall in the place of transition from output opening (22) into channel (30) forms edge (32). Compartment also includes subject section (50) for reception of output liquid, containing said cells and/or particles, and absorbing means (40), located adjacently to subject section (50) between channel (30) and subject section (50). Absorbing means (40) has opening (42) , making it possible for liquid, containing said cells and/or particles, move through opening (42) onto subject section (50), and additionally removes liquid from liquid, containing said cells and/or particles, on subject section (50) in such a way as to leave said cells and/or particles on subject section (50) for analysis.

EFFECT: realisation of more efficient, reliable and high-quality making of preparations of cells and/or particles, contained in liquid.

25 cl, 14 dwg

FIELD: measurement equipment.

SUBSTANCE: invention relates to measurement of total gas content in non-traditional container rocks, such as gas-bearing container beds, which may be found in sedimentary rocks, volcanic or metamorphic rocks. The method includes stages of well drilling in the measurement range in a container bed to create a volume of drilling mud in annular space, which contains fragments of drilled rock and gas. At the same time the volume of annular space has the front edge and the rear edge, diversion of the front edge of the annular space volume so that entire volume of annular space is trapped in a degassing system for storage without its exposure to atmosphere, interruption of diversion of annular space volume after trapping of the front edge of annular space volume in the degassing system for storage in order to determine quantity of gas in terms of annular volume; and also in-situ calculation of the total gas volume in the container bed with account of gas and fragments of drilled rock in terms of fragments of drilled rock and gas, contained in the annular space volume.

EFFECT: increased reliability and accuracy of the method and the device for measurement of total gas content in non-traditional container rock.

25 cl, 2 dwg

FIELD: automatical aids for sampling liquids.

SUBSTANCE: system for sampling and delivering filtrate has filter submerged into tested medium and connected with collecting tank and vacuum pressure source which is connected with top hole of collecting tank by means of pneumatic pipe. System has sample receiving tank connected with collecting tank and control unit which has first output to be connected with vacuum pressure source. Collecting tank has two separated chambers - washing chamber and dispatching chamber. Lower hole of washing chamber has to be lower hole of collecting tank and side hole of dispatching chamber has to be side hole of collecting tank. Floating valve is installed inside washing chamber to shut off lower and top holes. Filter is connected with lower hole of collecting tank through sampling pipe. Side hole of collecting tank is connected with lower hole of tank for receiving samples through sampling pipe. Flow-type sensor and check valve are installed inside transportation pipe. Output of flow-type sensor is connected with input of control unit; second output of control unit is connected with control input of analyzer.

EFFECT: improved precision of measurement of sample ion composition; prolonged service life of filter.

1 cl, 1 dwg

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