Method for additional electron-dense contrast enhancement of nucleic acids in nucleus and cytoplasm accompanying histochemical detection of sodium cations in cell and tissue ultrastructures
SUBSTANCE: invention refers to medicine and biology, namely to a method for additional electron-dense contrast enhancement of acid groups of biomolecules accompanying the histochemical detection of sodium cations in pulmonary and tracheal cell and tissue ultrastructures. Substance of the method consists in fixing tissue slices, washing the surface in bidistilled water, placing into a diluted agent containing 4% osmium tetraoxide 1 ml and 2% potassium hexahydroantimonate 8 ml. The tissue slices are stained for 4 hours while stirring strongly and washed in bidistilled water. The tissue slices are further prepared by slicing thinner and dehydrated; the semi-thin and ultrathin slices are produced and studied by a transmission electron microscope, which is followed by computer processing to detect diffuse selective staining of the acid cell ultrastructures and intracellular substance.
EFFECT: using the declared method enables the additional electron-dense contrast enhancement of the acid groups of biomolecules accompanying the histochemical detection of sodium cations in the pulmonary and tracheal cell and tissue ultrastructures.
5 dwg, 1 ex
The invention relates to the field of medicine and biology, histological, cytological, physiological study of tissues and cells of human organs and animals, as well as by cytological and histological methods.
Known methods histochemical staining of sodium chloride tissues and cells bodies, which is carried out using a saturated aqueous solution hexahydroquinoline potassium (K[Sb(OH)6]), a 2% aqueous solution of OsO4/G. Geier. E-histochemistry. - M.: Mir. - 1974. - 488 p/. Methods of staining cells with an aqueous solution of antimonate and osmium acid required for the study of biological objects with an electron microscope, since the products of biological staining structures containing sodium ions, these methods become electron-dense.
The excess of osmium tetroxide prevents this histochemical reaction, as phospholipids containing polyunsaturated fatty acids, have high electron density. Additional contrasting nucleic acids contained in the nucleus and cytoplasm of the cells uranylacetate, citrate of lead, etc., as well prevents histochemical detection of sodium ions through antimonate. Therefore, the main drawback of these methods is that they are made without with�of NASA coordination compounds OsO 4(osmium acid) with hexahydrocannabinol potassium, which occurs during heating of the aqueous solution of these substances in a water bath and subsequent purification dissolved when heated coordination compounds OsO4in water, acetone excess of osmium tetroxide, which is poorly soluble in water and in organic solvents.
Loved ones are essentially ways histochemical detection of sodium chloride and calcium ions in cells and tissues, which are made using antimonate /Komnik H. Electron mikroscopische Lokalisation von Na+ and CI - in Zellen and Geweben // Protoplasma. - 1962. - V. 55. - P. 414-418. Mariani P., Tolomio C, Baldan B., and P. Braghetta Cell wall ultrastructure and cation localization in some benthic marine algae // Phycologia. - 1990. - Vol.29. - No. 2. - P. 253-262/.
The significant difference of the claimed method from this is that it is the synthesis and purification: 1) main coordination compounds of osmium tetroxide, which are required for histochemical staining of cells and tissues, and 2) K2CO3that required to neutralize dye solution. Is used for this in eightfold molar ratio to the osmium acid 0.1 M solution hexahydroquinoline potassium (K[Sb(OH)6]). While there is no osmiophilic cytoplasmic membranes, if the duration of the staining for 4 hours or more.
Similar technical results�tattoo are ways histochemical staining of fixed coordination compounds of osmium tetroxide with timelinelite, dimethylaminoacetyl, unbalanced dimethylethylenediamine, tetramethylethylenediamine, tetraethylethylenediamine, which are used for histochemical detection of substances with an acid reaction (nuclear chromatin, nucleolus, ribosomes, glycosaminoglycans, etc.) in tissues and cells /Geier G. E histochemistry. - M. Mir. 1974. - 488 p/.
The significant difference of the claimed method from this is that as the ligand is used a saturated aqueous solution hexahydroquinoline potassium (K[Sb(OH)6]), resulting in synthesized Os[Sb(OH)6]4and K2CO3and what for histochemical detection of sodium chloride, calcium ions, inorganic ions of the iron used K[Sb(OH)6].
Close according to an embodiment is a method of producing pyroantimonate mercury (II) /Servinski A. E. EN 2036149 C1/, which is used in inorganic chemistry in the formation of compounds of antimony and mercury. The inventive method: the solutions of acetate of mercury (II) and potassium hydroxide is added to a solution hexahydroquinoline potassium. The molar ratio of K[Sb(OH)6]:Hg(CH3COO)2:KOH=2:(2-3):(6-8). Was stirred and heated for 10-15 min. the Precipitate was separated, washed with hot water and air dried at 100-120°C. the drying Time of 15-60 min. the Essential difference of the claimed method from this is �the formation of the coordination compounds of osmium tetroxide - Os[Sb(OH)6]4and K2CO3finally formed in the presence of NaCl and sodium ions (Na+), calcium (Ca+) found in the investigated organs and tissues of the body.
The task of the claimed method is the additional okrashivanie nucleic acids in the nucleus and cytoplasm, which is performed by electroncapture contrasting cells required for histochemical detection of sodium cations in ultrastructure.
The essence of the claimed method is that using a transmission electron microscope are studied in ultrathin sections of tissues and cells, fixed in histological fixative for 1 hour, contains 2% aqueous solution hexahydroquinoline potassium, and then with the aim histochemical detection of phosphate groups contained in nucleic acids, are painted with an aqueous solution of the basic coordination compounds of osmium tetroxide - Os[Sb(OH)6]4containing K2CO3that finally formed in the presence of NaCl and sodium ions (Na+), calcium (Ca+) found in the investigated organs and tissues of the body. The reaction of histochemical staining occurs at a molar ratio of the reactants inside the pieces of tissues containing sodium chloride etc. and reagents in the incubation with�food K[Sb(OH) 6]:OsO4that is equal to 1:(1-8):(1-10).
Our proposed method involves the synthesis of coordination compounds of the main interaction of osmium OsO4and osmium acid with hexahydrocannabinol potassium, which finally occurs in tissues, organs, in cells which contains in access of atmospheric air CO2that is absorbed in K2O. After fixation, slices of organs in histological fixative containing hexahydrocannabinol potassium, is the staining of the slices of the main bodies of the coordination compound of osmium, washing in distilled water, dehydrated in alcohol or acetone, opinion pieces in Araldite-EPON, fabrication of ultrathin sections, the study of tissues and cells using transmission electron microscope, computer processing of electronography.
The invention is illustrated: Fig. No. 1 - Chondrocytes fibro-cartilaginous membrane of the trachea of the rat. In the cytoplasm of chondrocytes revealed acidic groups of glycosoaminoglycans. Colour is the principal coordination compound of osmium, Fig.2 - Respiratory Department of lungs of the rat. Cellular elements of the interalveolar septum, selective staining of heterochromatin, euchromatin, nucleolus, perichromatin, interchromatin pellets, the main coordinating compound of osmium, Fi�.3 - The wall of the alveoli of the lungs of rats. In'veolocity detected electron-dense granules of calcium dihydrogen phosphate. In the intercellular space of the deposition of diffuse electroncapture material of sodium dihydrogen phosphate, Fig.4 - Intense contrast enhancement of the coordination compound of osmium cytoplasm and nuclei of the connective tissue cells of the interalveolar septum. In the intercellular space of the observed fine-grained precipitate antimonate sodium, Fig.5 - Deposition of sodium dihydrogen phosphate surfactant in the contingency. Ultrathin section of the lungs of rats.
The method is carried out in three stages, following each other:
1. At the beginning of the method to a 2% aqueous solution of OsO4in an eightfold molar ratio of added saturated 2% (0.1 M) aqueous solution hexahydroquinoline potassium K[Sb(OH)6]. Pre-mix the solutions individually for 30 minutes and heated in a water bath at 100°C. Then the solutions are mixed, and then gradually cooled with constant vigorous shaking to ensure access of atmospheric air rich in CO2that is absorbed in K2O.
2. For the purpose of chemical coagulation of the proteins, small (5 mm thick, 5 mm long) pieces of tissue of human or animal are recorded in histological fixative (glutaraldehyde) for 1 hour, then at�OSU 2% aqueous solution hexahydroquinoline potassium adjusted values of a latch to a pH of 7.4 (added dropwise). After that, the surface of pieces of bodies quickly washed from the first latch in bidistilled water and then in 2% aqueous solution hexahydroquinoline potassium. Then for 4 hours or more, pieces of tissues are stained in 4% solution of the basic coordination compounds of osmium and K2CO3bidistilled water. Staining is performed with constant vigorous shaking to ensure access of atmospheric air rich in CO2that is absorbed in K2O. After staining the tissue pieces prepariruetsya by cutting on a thin (1-2 mm thick, 2-2. 5 mm long), then rinsed in bidistilled water, dehydrated in ethyl alcohol or acetone and poured into araldit-araldit.
3. At this stage, on the instrument are made of ultra-thin tissue sections, which are studied using transmission electron microscope with subsequent computer processing of electronography with 3D graphics.
An example of use.
The proposed method further electroncapture contrasting acidic groups in biomolecules in histochemical detection of sodium cations in ultrastructure cells and tissues was used for histochemical studies of the respiratory Department of lungs and trachea intact rats. Malen�Chia (5 mm thick and 10 mm long) pieces of the tissues of the lungs and trachea were fixed for 1 hour in 4% solution of glutaraldehyde, to which was added dropwise 2% hexahydrocannabinol potassium in order to achieve a physiological pH of a solution, equal to 7.4. Quickly washed first in bidistilled water and then in 2% aqueous solution hexahydroquinoline potassium. Two slices (5 mm thick and 5 mm long) was placed in a reagent solution, which contained 1 ml of 4% osmium tetroxide and 8 ml of 2% hexahydrocannabinol potassium. The volume of pieces of organs empirically determined that the concentration of sodium chloride and cations of sodium, calcium, etc. in the body is approximately equal to 0.1 M. for 4 hours the pieces of tissues stained with vigorous shaking in 2% solution of the basic coordination compounds of osmium with hexahydroquinoline potassium bidistilled water. After staining, the slices quickly rinsed in bidistilled water. Then the tissue pieces prepariruetsya by cutting on a thin (1-2 mm thick, 2-2. 5 mm long). For the purpose of dewatering the tissue pieces are held through a series of alcohols, acetone and then poured into Araldite-EPON. Then on the ultramicrotome LKB NOVA were made of semi-thin and ultrathin sections.
In the case study of semi-thin sections of fibrous (fibrous)-cartilaginous membrane of the trachea and lungs is detected by the high-intensity granular staining of the cytoplasm of chondrocytes and chondrogenesis�lastow, cells of the alveoli and interalveolar septa, which is rich in glycosaminoglycans (Fig.1).
In the study of ultrathin sections of the lungs with a transmission electron microscope FEI Tecnai G2 Spirit was revealed diffuse selective staining of acidic cellular ultrastructure and intercellular substance, and the lack osmiofilii cytoplasmic membranes. In the nuclei of cells detected by selective staining of heterochromatin, euchromatin, nucleolus, perichromatin, interchromatin granules, which are characterized by high content of ions of phosphoric acid
In the cytoplasm of cells of the alveoli is detected weakly colored or moderately electronmobility diffuse material, faint granular material of 0.01-0.05 μm and a single electronmobility granules of calcium dihydroorotate Ca(H2PO4)2the size of 0.09-0.3 mm (Fig.3). In the cytoplasm of erythrocytes contained in the capillaries of the lungs, observed weak staining of the cytoplasm, found very small electron-dense granules of size 0.01-0.02 μm (Fig.2).
The conclusion about the presence of inclusions of calcium dihydroorotate Ca(H2PO4)2in lung cells and red blood cells made on the basis of previously conducted histochemical studies of the lungs of rats, which was carried on with�OSU alizarin red C /S. V. Zinoviev Histochemical characteristics of the venous system of the respiratory Department of lungs of experimental animals subjected to chronic hypothermia, after introduction into the body of dihydroquercetin // Bulletin of physiology pathology of respiration. - No. 45. S. 57-61/. In the intercellular substance in the cytoplasm and wall aeroheating barrier detects accumulation of neutral lipids, which are adjacent to diffuse accumulations electroncapture material size of 1-4 μm (Fig.2, 3).
On the surface of'veolocity first type is present diffusely painted a layer of surfactant which has a low electron density. In the surfactant varies two layers of gipofiz. On the surface of the surfactant are single very fine granules osmiophilic material (Fig.2, 3).
In the alveoli found colored coordination compound of osmium material, disordered located between subcellular structures in the intercellular substance in the contingency surfactant, in accumulations of neutral lipids, which contains molecules of sodium dehydroacetate (NaH2PO4) with moderate electron density, represented by a granule size of from 0.05 to 0.3 microns. The edges of these granules blurred, indistinct contours, some of them have a round or star-shaped (Fig.5).
Software Tr�nsmission electron microscope FEI Tecnai G2 Spirit allows you to carry out computer processing of electronography. When preprocessing electronography 3D graphics found crystals precipitate of antimonate, which contain sodium chloride and has a complex structure. With increasing electron microscope over 30,000 times, in the case of computer processing of electronography observed that the composition of the cytoplasmic vacuoles and in the intercellular space are detected fine-grained precipitate hexahydroquinoline potassium (0,0001-0,005 µm) with low or moderate electron density, which is formed by interaction with the sodium chloride (Fig.3, 4).
The method allows to synthesize, purify acetone basic coordination compounds of osmium - Os[Sb(OH)6]4excess of osmium tetroxide, which are used for histochemical detection of nucleic acids in the presence of NaCl and sodium ions (Na+), calcium (Ca+) found in the investigated organs and tissues of the body.
The technical result of the use of the method is that there is a qualitative staining of tissues, cells by inhibition of staining by osmium tetroxide phospholipids, with the aim of identifying nucleic acids (DNA and RNA), as well as sodium chloride solution, the cations of calcium, iron, etc., allowing you to spend significant ultrastructural study of ultrathin sections using Tr�nsmission electron microscope.
Thus, our proposed method can be used for electron microscopic studies of ultrastructure of cells and intercellular substance of organs and tissues of humans and animals, which have an acidic pH environment, Obnovlenie the presence of ions of phosphoric acid, sulfuric acid, and molecules of sodium chloride, or sodium cations.
The method further electroncapture contrasting acidic groups of the biomolecules in the histochemical detection of sodium cations in ultrastructure cells and tissues of the lungs and trachea, characterized by the fact that the pieces of 5 mm thick and 10 mm long tissue of the lung and trachea is fixed for 1 hour in 4% solution of glutaraldehyde, which was added dropwise 2% hexahydrocannabinol potassium to achieve a pH of 7.4, carry out cleaning of the surface from the retainer in bidistilled water and then in 2% aqueous solution hexahydroquinoline potassium, slices of 5 mm thickness and 5 mm in length is placed in a reagent solution,which contains 1 ml of 4% osmium tetroxide and 8 ml of 2% hexahydroquinoline potassium, for 4 hours, the tissue slices were stained with vigorous shaking in 2% solution of the basic coordination compounds of osmium with hexahydrocannabinol potassium bidistilled water, rinsed in bidistilled water, then the slices of tissue preparer�Ute by cutting on thinner 1-2 mm thickness 2-2,5 mm long, dehydrate, carried out through a series of alcohols, acetone and then poured into Araldite-EPON, and then on the instrument are made of semi-thin and ultrathin sections, which are examined with a transmission electron microscope, with subsequent computer processing of electronography 3D graphics with the detection of diffuse selective staining of acidic cellular ultrastructure and intercellular substance.
SUBSTANCE: preparation method of a dielectric specimen for investigation on a focused-beam electronic microscope of its micro- and nanostructure involves application of a current-carrying coating onto the specimen surface and provision of electrical contact of the specimen coating with the current-carrying object table. The current-carrying coating is applied by wetting of the specimen surface with a solution of hydrophilic non-evaporable non-flammable non-toxic current-carrying ionic liquid in the form of tetrachloroferrate of N-decylpyridinium in acetone and further drying of the specimen in the air till complete removal of a volatile component.
EFFECT: prevention of accumulation of electrical charges on the surface of dielectric specimens.
FIELD: oil and gas industry.
SUBSTANCE: sampling device contains a main pipeline, a sampling section joined with the main pipeline with a possibility of sampling with coverage of liquid flow cross section, a sampling tap and a manometer. The sampling tap is designed as housing and bushing which is rigidly joined with a handle, and in a starting position the bushing shuts a drain hole of the housing, and the handle can move together with the bushing, opening the drain hole of the housing in working position. The hollow cylinder with the central channel is installed in the main pipeline, and the hollow cylinder from side of liquid flow movement is fitted with the input cone tapering the flow and the output cone expanding the flow on the other side of the hollow cylinder. The whirler fitted inside with tangential channels is installed in the central channel of the hollow cylinder from the input cone. The bushing of the sampling tap covers hermetically from outside the housing with the drain hole and can move restrictedly in axial direction with reference to the housing. The housing has the first and second external cylindrical grooves and the cutting spring lock ring is located inside. The bushing is fitted with internal ring sampler.
EFFECT: improvement of quality of liquid sampling, improvement of operational reliability of the sampling tap and increase of level of correctness of identified process parameters of wells and layers according to analyses of samplings.
SUBSTANCE: method comprises the steps: collecting from the bile ducts of liver of domestic and/or wild animals infected with fasciolas of only live adult F. hepatica. Placement them into individual tubes with filtered and centrifuged bile diluted with isotonic solution of sodium chloride 1:1. Exposure of tubes at t = 38-39°C if F. hepatica is from cattle, and t = 39-40°C if F. hepatica is from sheep and/or goats, under conditions of thermostat for 5 hours. Subsequent washing eggs in isotonic solution of sodium chloride.
EFFECT: invention enables to obtain up to 100 percent of fertilised eggs of Fasciola of species Fasciola hepatica and can be used for study in the laboratory or field experiments in solving fundamental and applied scientific tasks in the field of epizootiology, treatment and prevention of fascioliasis of domestic or wild animals.
SUBSTANCE: invention relates to the field of agricultural machinery industry. The device for sampling chopped straw from grain combine harvesters comprises collectors, a retainer, a bracket and a control lever. The collectors are arranged in rows, the number of which in the transverse direction is determined by the header coverage and the selected number of the studied areas, and in the longitudinal - replication of sampling. The collectors are interconnected in longitudinal rows by means of flexible connections equal in magnitude, so that the total length of the row does not exceed the distance to the ground mass descent area on the stubble. The first collectors in each longitudinal row have easily removed connections on the holes in the rod-float leveller which is located perpendicular to the direction of the combine movement and retained by the retainer driven by the control lever.
EFFECT: invention provides a boundary separation of the comparable areas in sampling and reduction of the probability of failure of experimental equipment.
2 cl, 1 dwg
SUBSTANCE: invention relates to the method of analysis of a set of ferromagnetic particles. The method is characterized by that the particles of the named set are levelled in such a way that each of the named particles is oriented practically in the same direction. Then the particles of the named set are fixed in this aligned direction and the internal areas of the named particles levelled in such a way are uncovered. After that the nature of the alloy comprised by each of the named particles is identified, the named particles are grouped by categories depending on their nature and the metallurgical structure and chemical composition of one or more of the named particles in each category are determined.
EFFECT: improvement of accuracy and reliability of the analysis of ferromagnetic particles.
14 cl, 6 dwg
SUBSTANCE: invention relates to forecast of ageing processes of the synthetic polymer materials (SPM) depending on duration of their operation or storage. Analysis of the volatile organic compounds (VOC) migrating from SPM is performed by active sampling for sorbent, with further thermal desorption and gas chromatographic analysis. Forecast of the ageing processes of the material and estimation of toxicity of the gas discharge are performed as per dynamic of the qualitative and quantitative composition of gas discharge components in SPM initial state, and during artificial climatic thermal-humidity ageing. Dynamics analysis of the total gas discharge (ΣT) from each material is performed for all substances migrating from studied SPMs. Change of toxicity is estimated and forecast of the material ageing is performed as per developed indices of total gas discharge (ΣT) and as per hygienic index P=(ΣTinitial/ΣTn)/V, where Tinitial and Tn are indices of toxicity of gas discharge of each substance in initial state and after ageing, respectively, and ΣTinitial and ΣTn are total indices of toxicity of gas discharge of all components of SPM in initial state and after ageing, V is duration of ageing (year, month).
EFFECT: invention ensures high accuracy of the method of VOC qualitative and quantitative composition determination in gas discharge during materials ageing and the analysis results repeatability.
FIELD: physics, acoustics.
SUBSTANCE: group of inventions relates to an apparatus for irradiating a sample with focused acoustic energy, a device which is part of said apparatus, a cartridge for said device and a method of irradiating a sample with focused acoustic energy. The apparatus comprises a device, a cartridge, a completely solid-state connector and a source for generating acoustic energy. The cartridge has a chamber for receiving a sample, and the completely solid-state connector provides a completely dry coupling of acoustic energy between the source and the cartridge. The device and the cartridge are adapted for inserting the cartridge containing a sample into the device and are separable, and the focused acoustic energy is focused high-intensity ultrasound. The device has a source for generating acoustic energy and the cartridge has a chamber for receiving a sample.
EFFECT: improved sample processing.
17 cl, 24 dwg
FIELD: motors and pumps.
SUBSTANCE: simulation device contains an oil batcher, a dispersion chamber and a lubricant oil decomposition chamber (1). At the air outlet downstream the chamber the diffuser (2) is located. On the chamber the heater (3) with the thermocouple (4) and the thermorelay (5) is installed. The device includes the air duct (6) supplying the pumped-over hot air into the lubricant oil decomposition chamber connected through the manometer (7) to the air compressor (8). The device contains the cylinder (13) filled with ultra-pure nitrogen, (23, 24) the sealed gage tank with air cavity with oil and a cover for oil filling, with the oil pipeline connected to it through the gas pipeline with the regulator (12), the adapter (11) and cap nuts. The gage tank (9) is connected through the adapter (11) with cap nuts (20, 21) to the measured capillary (15) in a cooling jacket (16) with circulating water through the thermostat with the pump (18) and radiators, attached to the decomposition chamber by means of the cap nut (22) and the sealing cone (25). Also the device comprises the additional chamber (26) screwed to the main decomposition chamber of (1) coaxially and sealed with a gasket (27), with the rod with a flywheel (17) installed inside, with threaded and non-threaded parts. Meanwhile the threaded part is implemented with a possibility of movement in the internal washer with a thread (28) for adjustment of the decomposition chamber volume and change of conditions of simulation of oil concentration, while the non-threaded part is sealed in a gland with graphite seal (29).
EFFECT: improvement of accuracy of simulation of composition of oil decomposition products in aviation gas-turbine engines.
FIELD: aircraft engineering.
SUBSTANCE: invention relates to aircraft cabin air samplers, gadgets for analysis of admixtures in aircraft cabin air samples for analysis of concentration of contaminants in aircraft air conditioning systems and for determination of the composition of harmful admixtures and dangerous concentrations of gases and vapours in air. This sampler comprises evacuated vessel as air consumption booster, absorption cartridge with sorbent-concentrator composed of sharpened steel tube with plugs of glass wool at tube ends, with side bore in said tube filled with sorbent and glass wool. Evacuated vessel is composed of cylindrical case with inlet and outlet pipes fitted at case ends. Outlet pipe is provided with tube of vacuum rubber and metal plug to be fitted in the tube free end after evacuation. Inlet pipe is welded to the case end and features ID larger than that of the case inlet and internal thread. Aforesaid absorption cartridge with sorbent-concentrator is fitted in said inlet pipe and, partially, in evacuated vessel used also as a sampler of admixtures not absorbed by concentrator. Locking device composed of the tube with air sample passage inlet is screwed via seal ring in evacuated vessel surface neck to tight fit. Cover with stiffness ribs and rubber washer is secured to said locking device and aligned therewith. Lever with triangular cam at the end is articulated with the device on opposite side from the cover attachment side to lift said cover to unseal the system and to bleed air. Said lever and cover are secured to be revolved in one plane relative to axles of rotation and attachment. Springs secured from one side to locking device case and to cover stiffness ribs on opposite sides. This makes said cover opened and cover closed at lowered lever and cover located at lever can outer leg. Lever shifted, cover goes up to unseal the system and to bleed air.
EFFECT: higher sampler sensitivity, accuracy of analysis, accelerated in-flight experiment.
FIELD: oil and gas industry.
SUBSTANCE: system contains casing-well, cylindrical sampler comprising three main parts, top part is manifold chamber, middle part is connecting coupling with female thread and groove connecting bottom and top parts, bottom part is receiving chamber for accumulation of gas supplied via side holes of the casing-well, receiving chamber and manifold chamber are covered with lids, above the connecting coupling a discharge tube is installed, under it a receiving tube is installed, above it a ball valve is installed, top discharge tube passes through the manifold chamber, lid and goes outside, on it the inlet union valve is installed to inject air in the manifold chamber and safety union valve for overpressure relief, the pneumatic chambers are located one above, and another below the inlet holes in the sampler casing, in top lid of the sampler the outlet valve is installed, casing pipe is made out of n pipes connected by outside thread coupling, with side holes of same diameter uniformly distributed along length of the casing pipe-well.
EFFECT: simplified design.
SUBSTANCE: invention refers to medicine, particularly to haematology. The cryogenic preservation of haematopoietic umbilical cord stem cells is carried out with a cryoprotectant solution - dimethylsulphoxide - added to a suspension of nuclear cells with the haematopoietic stem cells. That is followed by preparing for freezing by cooling the stem cell suspension in a cooling chamber to a temperature of +4°C. The multi-staged freezing of the stem cell suspension involves using a cryoprotectant solution that is 55% dimethylsulphoxide with 5% dextrane 40, which is added into a suspension of a leukocyte concentrate with the stem cells packed in a cryobag. That is followed by mixing mechanically in a mixing apparatus and added with a cryogenic preserving agent at a temperature of +4°C, de-aerating the cryobag with releasing a portion of the suspension, closing the bag, sealing it and placing into a shrink bag and freezing within several stages with computer assistance. At the first stage, the stem cell suspension mixed with the cryoprotectant solution (hereinafter referred to as a sample) is kept for 10 min at a temperature of +4°C, frozen at 1°C/min to a temperature of -12°C, cooled down at 15°C/min to a temperature of -60°C; after the sample is unfrozen at 10°C/min, it is cooled down at 1°C/min to -60°C; at the end of the freezing programme, the sample is cooled down at 3°C to -100°C. Upon completion of the freezing programme, the sample placed into a cryobox is placed into a quarantine vacuum flask containing liquid nitrogen to determine test results for the presence or absence of infectious agents and bacteriological and mycotic contamination. At the end of the quarantine period, the haematopoietic umbilical cord stem cell sample is transported into the vacuum flask containing liquid nitrogen for long storage at min -150°C with negative test results.
EFFECT: invention enables increasing the cells viability.
1 tbl, 1 dwg
SUBSTANCE: invention relates to a solution for fixing biological cells. The fixing solution is intended for in vitro preservation of a cytological sample containing nucleated cells and erythrocytes. The solution contains 80-95 vol. % of a mixture of 590 ml normal saline, 10 ml polyethylene glycol (Carbowax®), 203 ml isopropyl alcohol, 193 ml pure ethanol, 0.01 vol. % sodium azide, and 5-20 vol. % buffered 4% formalin; pH of the fixing solution is in the range of 6.4 to 7.4.
EFFECT: preserving the integrity of nucleated cells.
2 cl, 2 dwg, 1 ex
SUBSTANCE: coloured X-ray contrast mass consists of barium sulphate, glycerine, acrylic dye and aqueous gelatine solution. The invention relates to methods of producing a setting coloured X-ray contrast mass. The disclosed mass can be used for macro- and micro-preparation of vessels and X-ray imaging thereof. The preparation method includes thoroughly mixing barium sulphate, glycerine, acrylic dye and aqueous gelatine solution, wherein barium sulphate, glycerine, acrylic dye and aqueous gelatine solution are taken in amount of 1.5 parts, 1 part, 0.1 parts and 7.5 parts, respectively, stirring and heating the mixture to 60-80°C before application. The prepared mass has high permeability in thin vessels, brightness (clarity), fast setting and elasticity. The mass is prepared from readily available and cheap materials. The remaining or unwanted mass can be stored for a long period of time.
EFFECT: disclosed mass can be heated repeatedly and is ready for use.
SUBSTANCE: cryoprotector is opened in a laminar flow unit; a special syringe of asyringe pump is filled with a cryoprotector; that is followed by introducing a filter solution of the cryoprotector - 55% dimethylsulphoxide with 5% dextran 40 at temperature +4°C in a nucleated cell suspension with haemopoietic stem cells in a cryopackage with the concentrate and mixing mechanically in a mixing apparatus, transferring the system together with the cryoprotector flask into the laminar flow unit; the air is released from the cryopackage and portion of the suspension; the package is sealed and placed into a shrink bag; that is followed by programmed multi-stage freezing, the first stage of which keeping the mixture of the suspension with the stem cells and cryoprotector - a freezing sample - for 10 min at temperature +4°C; the second stage is cooled at a rate of 1°C/min to temperature -12°C; the thirst stage provides cooling at a rate of 20°C/min to temperature -60°C; at the fourth stage, the sample is heated at a rate of 10°C/min to temperature -18°C; at the fifth stage, the sample is cooled at a rate of 1°C/min to -60°C; at the end of the freezing program, the sample is cooled at a rate of 3°C/min to temperature -100°C; after freezing, the sample is placed into a quarantine dewar with liquid nitrogen until infection and bacteriological fungal contamination test results are obtained. After termination of the quarantine shelf life, the sample with haemopoietic stem cells are placed for long-term storage at temperature not exceeding -150°C, into the dewar with liquid nitrogen if observing negative test results. If the infection and bacterial and/or fungal contamination test results are positive, the sample with haemopoietic stem cells are transferred into the dewar with liquid nitrogen for infectious material for long-term storage.
EFFECT: invention enables increasing cell viability in the sample.
3 cl, 4 dwg
SUBSTANCE: invention relates to method of obtaining 1,6-bis-(1,5,3-dithiazepan-3-yl)-2,5-disulphanylhexane of formula (1), possessing fungicidal activity against Botrytis cinerea and Rhizoctonia solani. Essence of method lies in interaction of mixture of 1,2-ethanedithiol and formaldehyde with water solution of ammonium salts NH4X (X=F, Br, OAc, NO3, 1/2SO4) with molar ratio HS(CH2)2SH:CH2O:NH4X=3:6:2 at room temperature (~20°C) and atmospheric pressure for 5-7 h.
EFFECT: output of 1,6-bis-(1,5,3-dithiazepan-3-yl)-2,5-disulphanylhexane of general formula (1) depending on applied ammonium salts (NH4X) constitutes 31-67%.
2 cl, 2 tbl, 1 ex
SUBSTANCE: paleontological objects with soft tissues are treated for 1-4 months with an embalming solution containing 40% formalin - 0.516%, crystalline phenol - 0.262%, glycerine - 79.077%, 96% alcohol - 18.576%, sodium chloride - 1.569%. The ratio of the embalming solution to the mass of the paleontological object must not be less than 3:1. Embalming is carried out at room temperature.
EFFECT: method of embalming paleontological objects with soft tissues provides prolonged preservation of soft tissues, minimises loss of soft tissues having different rotting stages, stops rotting processes, skin straightening, preserves natural colour thereof and natural morphometric parameters, which enables further use of the paleontological object for scientific purposes and for exhibition in dry form.
SUBSTANCE: invention refers to physiology, cryobiology and medicine, namely to human blood nuclear cell preservation technique with the use of inert gas. The method involves cell saturation in Compoplast 300 plasticised container placed in a metal cryobarocontainer with inert gas, xenon at pressure 0.6 atm for 20min in the room temperature environment of 21±2°C; excessive gas is eliminated, and the biological object is removed from the metal container and frozen to -28°C in ethanol, placed into an electric freezer at -80°C. The bioobject is unfrozen in a water bath at 39±1.5°C for 1.5 min.
EFFECT: implementing the invention provides the reliable cryonic preservation of leukocytes in the inert gas medium and the high level of qualitative and morphological safety of leukocyte concentrate of human blood.
1 tbl, 3 ex
SUBSTANCE: method of reducing impact of low-temperature jump on cryoprotectant solution is provided at the expense of remote processing of cryoprotectant solution with cells of living organisms by ultrasonic radiation with frequency 0.50-10 before its freezing.
EFFECT: reduction of low-temperature jump of cryoprotectant solutions, which makes it possible to eliminate overcooling effect, provide continuous and smooth character of freezing and increase integrity of defrosted cells after cryoconservation.
SUBSTANCE: what is presented for embalming of dead bodies for the purpose of organ harvesting and transplantation drills is a method, wherein tissues and organs are saturated with embalming solutions; the dead body is immersed into water at t 40°C for 120-160 min; cannulas are inserted into a femoral artery, and a blood flow is washed with warm normal saline with 1-2% sodium citrate added; the femoral artery is sealed, and a solution containing formalin and glycerol is inserted into the artery.
EFFECT: keeping the internal organ structure by increasing the preserving properties of the solution and providing the small vessel filling.
SUBSTANCE: invention relates to a method of lyophilisation of a composition, which contains purified antithrombin III (AT III) and a crystallised substance, selected from alanine, mannitol, glycine or NaCl. The claimed method includes freezing the composition at a temperature from -52°C to -60°C for 6-15 hours, annealing the composition at -30°C for 1 hour, re-freezing the composition at a temperature from -52°C to -60°C for 2-15 hours at keeping the product temperature between -48°C and -52.7°C for 4-10 before lyophilisation and drying the composition with obtaining a lyophilised cake. The invention also relates to a pharmaceutical set, which contains the said lyophilised cake and a liquid reagent.
EFFECT: invention provides obtaining the lyophilised composition, containing AT III, which preserves its activity and stability.
14 cl, 24 dwg, 5 tbl, 2 ex
SUBSTANCE: method involves using product containing spermatozoa treated by drying with freezing to humidity level of 1% and having injured membrane or spermatozoon head so that spermatozoon nucleus retains its genetic validity enough for fertilization. The spermatozoon heads fertilize an isolated oocyte after rehydration and microinjection being done. The retained genetic integrity is enough for fertilizing an oocyte and producing living descendants. Method involves collecting living mature spermatozoa, making spermatozoa suspension in special purpose physiological medium, freezing the spermatozoa suspension for producing frozen spermatozoa, drying the frozen spermatozoa or spermatozoa heads in vacuum to humidity level of 1%, making rehydration of spermatozoa or spermatozoa heads with injured membrane and selecting those retaining nucleus of genetic integrity. The selected spermatozoa or spermatozoa heads are used for fertilizing isolated oocytes with living descendants being produced.
EFFECT: enhanced effectiveness in producing living descendants.
39 cl, 5 dwg, 1 tbl