Method of preparing bone tissue preparation and set for its realisation
SUBSTANCE: invention relates to medicine, veterinary and biology. Sample fixation, decalcification, washing with water, dehydration in alcohol solutions and pouring with paraffin are performed. Sample fixation is performed for 24 hours in alcohol-based molecular fixing agent FineFix, which contains FineFix and 96° alcohol in ratio 1:2.5. Decalcification is carried out for 2-5 days in 5-8% buffered solution of formic acid with daily replacement of decalcifying solution and control of decalcification completeness. Ratio sample:decalcifying solution constitutes 1:20. When decalcification is finished, sample washing with water is performed and sample is re-submerged into alcohol solution of molecular fixing agent FineFix for 6-12 hours before dehydration stage. Set for preparation of bone tissue preparation contains alcohol-based molecular fixing agent FineFix, concentrated solution of decalcifying agent, prepared with ratio 40 g of sodium citrate, 100 ml of 90% solution of formic acid, 300 ml of distilled water, and working solutions for control of decalcification completeness, containing saturated ammonium oxalate solution and 25% water solution of ammonia.
EFFECT: invention makes it possible to obtain high-quality preparations suitable for further histological and immunohistological analysis without application of highly toxic components.
2 cl, 2 dwg
The present invention relates to the field of medicine, veterinary medicine and biology.
Histological examination of bone tissue is a required component diagnosis of many diseases and conditions and is used in clinical and experimental studies. The quality of the product plays a crucial role in diagnosis and in establishing the fabric of the relationship.
There is a method of preparing bone, including bone cutting and grinding (Microscopic techniques / Ed. by Sarkisov D.S. Petrov JUL - M.: Medicine, 1996. - S).
The disadvantages of this method include the fact that the resulting preparations are unsuitable for histological studies, as in thick slices obscured the structure of the bone, and when grinding also destroyed cellular elements.
Also known a method of preparation of fragments of the bone tissue to the research, including fixation in buffered 10%solution of neutral formalin for 24-48 hours, rinse in water for 24-48 hours, declinatio 5-7 .5%solution of nitric acid to the full decalcomania, subsequent processing of the preparation of bone tissue in 5%solution of sodium sulfate or lithium within 1 day, washing in water for 24-48 hours, dehydrated in alcohols and fill in paraffin (ICRI is scopacasa technique / edited Sarkisov D.S. Petrov JUL - M.: Medicine, 1996. - S, 457, 467).
However, this method does not allow to achieve good preservation of cellular structures, especially in the surrounding soft tissues (muscle, periosteum, bone marrow), significantly affects the quality standard histologische colors, and does not allow subsequent immunohistochemical and immunofluorescence studies due to the damage of the secondary structure of protein epitopes.
The closest in technical essence to the present invention is a method of preparation of bone samples for morphological diagnosis, including the fixation of the sample, its declinatio, washing with water, dehydration in ethanol solutions with increasing concentration and fill in paraffin (Medvedev N.N., Nikolaev V.G. and other Method of preparation of bone samples for morphological diagnosis. Pat. EN 2241994, IPC7G01N 33/48, G01N 1/28, Application: 2003129253/15, 30.09.2003, Published: 10.12.2004).
The known method is as follows. After fixation of bone samples in formalin solution are declinatio within 7-8 days in a 5%solution of nitric acid on the basis of a 5%formalin solution, after which bone samples for 2 hours, washed under running tap water and 12 hours immersed in a 5%solution of sulfuric acid lithium, then for 20-30 mine is again washed with running water and subjected to dehydration in alcohols of increasing concentration.
The disadvantages of this method include the duration of the procedure of decalcomania, use as a fixative formalin, and as calcinator solution of nitric acid, which prevents the subsequent conduct immunohistochemical and immunofluorescence studies of samples due to the significant damage to the surrounding tissue and cellular structures when conducting decalcomania.
The task of the invention is to develop a method of preparing bone, suitable for histological and immunomorphological research, and set for its implementation.
The technical result of the proposed method is the preservation of the histological structures of the bone and surrounding tissues, as well as reducing the complexity and toxic effects in the manufacture of the drug.
The technical result is achieved in that the method of preparation of preparation of bone tissue includes the fixation of the sample, its declinatio, washing with water, dehydration in ethanol solutions and fill in paraffin.
The difference of the proposed method lies in the fact that the fixation of the sample bone should be performed within 24 hours in molecular clamp, which does not include aldehydes, for example, raster FineFix alcohol-based, containing FineFix is 96° ethyl alcohol in the ratio of 1:2,5.
FineFix - Besfamilnov the latch, which includes the following components: water, 1,2-PROPANEDIOL, polyvinyl alcohol and from 0.05 to 2.00 wt.-% Monomeric carbohydrate containing at least 3 carbon atoms and the second means for storing ethanol (patent EP 145 5174 B1 Fixative, the application Number EP20030005010, publication date: December 15, 2004, priority date: March 5, 2003. Other numbers of patent: US 20050074422. Author: Francesco Visinoni, The Applicant Is A Milestone S.r.L).
Distinctive technique proposed method is that declinatio carried out in 2-5 days 5-8% buffered solution of formic acid at a daily changing decalcifying solution and verify the completeness decalcomanie ratio sample:decalcifying solution is 1:20.
The differences between the proposed method also includes receiving repeated premises of the sample bone in alcohol solution molecular retainer for 6-12 hours, which is carried out after decalcomania and washing the sample with water.
Comparative analysis of the proposed technical solutions to the prototype allows to make a conclusion on the conformity of the proposed technical solution the criteria of the invention of "novelty."
Distinctive techniques of the proposed method ensures the safety of histological structures of the bone and surrounding tissues, preservation of antigenic epitopes,and reduce labor intensity and reduce the time of manufacture of the drug. So, the timing of decalcomanie compact bone small laboratory animals (mice, rats) are 2-4 days, rabbits 4-5 days, fragments of human bone tissue - 4-5 days. This eliminates the toxic effects of formalin on the researcher.
The inventive method achieves perceived by the applicant of the technical result, namely, the preservation of the histological structures of the bone and surrounding tissue, reducing the complexity and faster manufacturing of drugs.
Therefore, the inventive method allows to obtain preparations of high quality, suitable for routine histological examination and for carrying out immunohistological studies. In addition eliminates the use of highly toxic formalin.
The above can conclude that the technical solutions according to the criterion of "inventive step".
The method constituting the invention, intended for use in medicine, veterinary medicine and biology. The possibility of its fulfillment is confirmed as described in the application techniques and equipment.
The above gives grounds to believe that the proposed solution meets the criteria of the invention "industrial applicability".
Implementation of the proposed method is illustrated by a specific example is on the run. The following example serves to illustrate, but not limit the invention.
Samples of bone tissue fixed in alcohol solution molecular latch FineFix (280 ml FineFix+720 ml of 96° alcohol) for 24 hours. Then without washing the sample is placed in an individual well-closed container in a 5%buffered solution of formic acid. A concentrated solution of calcinator, the so-called matrix solution, at the rate of 40 g sodium citrate, 100 ml of 90% formic acid, 300 ml of distilled water; before use the solution was adjusted with distilled water to obtain a 5-8% solution of formic acid. For each sample the ratio of the sample:decalcifying solution is 1:20.
Daily assessment of the completeness of decalcomanie calcium-oxalate method (3 ml used decalcifying fluid, to bring the pH to 7.0 with concentrated (25%) ammonia solution, then add 5 ml of saturated ammonium oxalate solution, mix well, leave on for 30 minutes while the liquid remains clear - decalcomania completed). In the presence of calcium used in decalcifying solution, the sample is placed in a fresh portion of calcinator.
After decalcomanie carry out washing of the samples with water for 5 minutes and then placed in alcohol solution FneFix 12 hours.
This was followed by dehydration in alcohols of increasing concentration ranging from 70° solution and ending at 100° alcohol. Fill in paraffin carried out according to the standard technique (Microscopic techniques / Ed. by Sarkisov D.S. Petrov JUL - M.: Medicine, 1996. - S).
The proposed method produced 162 block of samples of bone tissue. Obtained high-quality histological preparations as the application of routine methods of staining (staining with hematoxylin-eosin)and when applying immunomorphological methods of dyeing.
When the preparation of bone tissue using a set containing molecular latch FineFix alcohol-based, containing FineFix and 96° ethyl alcohol in the ratio of 1:2.5, the concentrated solution of calcinating and working solutions to monitor the completeness of decalcomanie: 25% ammonia solution and a saturated solution of ammonium oxalate.
The essence of the proposed method is illustrated in figures histological preparations of bone tissue, obtained by the proposed method.
Thus, figure 1 shows the histological preparation of the bone, stained hematoxylin-eosin. Shows the security structures of the bone and surrounding tissues And muscles, periosteum, bone, D - bone marrow, E - cartilage, 40x magnification.
Figure 2 shows the staining of bone marrow cells in detalizirovan the th bone immunohistochemical method on carbonic anhydrase. Used primary antibody CA2, Epitomics® catalog No. 5204-1. Demonstrated good quality staining. F - brightly coloured osteoclasts, D - bone, magnification 400x.
Thus, the proposed method allows to obtain preparations of bone tissue, suitable for histological and immunomorphological research. When this bone and the surrounding tissue to maintain its structure, are preserved antigenic epitopes, there is no use of highly toxic components.
1. The method of preparation of preparation of bone tissue, including the fixation of the sample, decalcomania, washing with water, dehydration in ethanol solutions and fill in paraffin, characterized in that the fixation of the sample should be performed within 24 h in the molecular latch FineFix alcohol-based, containing FineFix and 96%ethanol in the ratio 1:2,5, declinatio carried out in 2-5 days 5-8% buffered solution of formic acid at a daily changing decalcifying solution and verify the completeness decalcomanie ratio sample:decalcifying solution is 1:20, after decalcomanie spend washing the sample with water and to the stage of dehydration re-place the sample in alcohol solution molecular latch FineFix 6-12 hours
2. Kit for the preparation of the preparation of bone tissue, characterized the decomposing those it contains a molecular clamp FineFix alcohol-based, concentrated solution of calcinator made at the rate of 40 g sodium citrate, 100 ml of 90%formic acid, 300 ml of distilled water and working solutions to monitor the completeness of decalcomania: a saturated solution of ammonium oxalate and 25%aqueous ammonia solution.
SUBSTANCE: compound for conclusion of histological preparations contains synthetic resin, solvent and auxiliary component. Foamed polyurethane (FPU) is used as resin, xylol is used as solvent and dibutyl phthalate is used as auxiliary component at the following ingredient ratio, wt %: foamed polyurethane 15-20; xylol 79-84; dibutyl phthalate 1.0.
EFFECT: reduction of cost of embedding medium and improving quality of histological preparation at maintaining fabrics colour.
3 tbl, 3 ex
SUBSTANCE: plastic deformation is created on edges of a specimen. The following is used for creation of plastic deformations: local heating, rolling and combination of those methods of action; as a result, in the centre of the specimen a zone with uniform distribution of stress components appears, the signs and levels of which depend on the nature of action and can be found as per the measurement results of the specimen before and after the action.
EFFECT: improving accuracy of measurement results of mechanical stresses in steel structures.
2 cl, 2 dwg
SUBSTANCE: method for preparing biological tissue sample for histopathological examination wherein the biological tissue samples are sequentially processed in a fixative solution which is presented by 10% formalin buffered by sodium phosphates; the intermediate processing takes place at room temperature. Then, the biological tissue samples are washed with water. That is followed by dehydration in more than one portions of isopropanol. The samples are cleared in two portions of the intermediates containing isopropanol and mineral oil with the first portion of the intermediate containing five portions of isopropanol and one portion of mineral oil, and with the second portion containing two portions of isopropanol and one portion of mineral oil. The intermediate processing is followed by immersing the biological tissue samples into mineral oil for clearing. The biological tissue samples are impregnated with melted paraffin added with 5% of bee wax, 0.8% of dimethyl sulphoxide and 0.5% of butyl rubber.
EFFECT: reducing labour content and time for preparing the high-quality section of the histopathological examinations with the either laboratory facilities: manual and automatic lines, in any tissue processors due to the high quality biological tissue samples having been prepared.
SUBSTANCE: first, a specimen of rectangular shape is made; a bevelled cut is made on the specimen at an angle of 15-25° from lower base to upper one, thus taking specimen length as the base. Then, surface of the bevelled cut of the specimen is ground and an austenisation mode is performed in oxidation atmosphere by etching gas. The specimen is cooled in water or in the air; then, a microslice or a series of microslices is prepared on surface of small base of the specimen, thus grinding the layers parallel to large base of the specimen. Depth of decoration zone is measured by etching gas on microslice by means of a microscope; then, the investigated surface of microslice is etched with alcohol solution of nitric acid till boundaries of austenitic grains are determined; determined boundaries of austenitic grains are studied; depth of selective determination zone of boundaries of austenitic grains is determined, and the defined etching picture is photographed. As per investigation results of microslice surface, structural state of the specimen is evaluated step by step: first, decoration zone of structure by oxidation with etching gas, then, selective etching zone of actual boundaries of austenite grains, and finally, zone of simultaneous detection of boundaries and intra-grain structure of the investigated steel; then, full depth of penetration of etching gas to the investigated material is determined by summing depths of decoration zone with etching gas and zone of selective detection of boundaries of austenite grains at etching of microslice and multiplication of the obtained value by cosine of inclination angle of bevelled cut to the large base.
EFFECT: simpler detection of boundaries of actual austenite grain; providing complex evaluation of structural state of hardened steel with possibility of multiple layer-by-layer investigation of slices by simultaneous fixation of oxidation zone of the investigated steel, zone of selective detection of boundaries of actual austenite grain and zone of intra-grain structure on slice surface.
7 dwg, 2 tbl
FIELD: testing equipment.
SUBSTANCE: invention is designed for sampling and precise comprehensive assessment of pollution of air samples (from air supplied into a system of aircraft pilot cabin conditioning), sampled from a compressor of a gas turbine aviation engine (GTE) during its bench testing, and further gas chromatographic analysis of samples for content of hazardous admixtures. A laboratory complex for sampling and gas chromatographic analysis of air samples includes a complex of air sampling 6 with a unit of samplers 7 and a control panel 8, a complex for gas chromatographic analysis of air samples 3 with a control panel 4, a package 9 for transportation of adsorption packets 10 and a container 11 for storage of concentrators 12. The laboratory complex is also equipped with plants for supply of gases 5, pumping of calibrating gas mixture 2 and definition of working volumes of a vacuumised part of the item 1. At the same time the plant for supply of gases 5 is simultaneously connected with the plant for pumping of the calibrating gas mixture 2 and with the complex of gas chromatographic analysis of air samples 3, and the plant for definition of working volumes of the vacuumised part of the item 1 is connected with the complex of air sampling 6.
EFFECT: improved operational properties, provision of sampling and precise comprehensive assessment, total error of measurement of pollution of air samples from a GTE compressor under bench testing, higher quality of indirect control of oil seals used in supports of the GTE compressor rotor.
FIELD: testing equipment.
SUBSTANCE: sampler relates to a sampling device in liquid and fluid condition, namely, to samplers for semi-automatic sampling along the entire height of the reservoir with oil products. The sectional sampler comprises a sampling column, a system of three-way valve control in the form of connecting traction rods and yokes connected into a parallelogram. Sections of sampling pipes are assembled by means of their installation into bores of three-way valves and are fixed by captive nuts with a seal. Besides, a worm gear is installed on the axis connected to a master yoke.
EFFECT: higher operational characteristics and service life of a sampler due to reduction of labour intensiveness of its assembly during installation and repair.
SUBSTANCE: invention relates to field of medicine, namely to pathomorphological diagnostics. To predict five-year long survival of patients with invasive breast cancer index of dispersion of tissue structures is determined as difference between maximal and minimal values of number of cancer structures and/or fractions of parenchymal or stromal component in microscopy at small magnification (100x) divided by quantity of vision fields in which said values were counted. If index of dispersion of tissue structures is lower than 1.6, five-year long survival of patient can be predicted with 95% probability, if its value is higher than 2.3, prediction of five-year long survival is unfavourable.
EFFECT: method makes it possible to predict five-year long survival in patients with invasive breast cancer.
FIELD: machine building.
SUBSTANCE: device includes sampling tube mounted in pipeline perpendicular to flow movement and provided with slot-like inlet from side of flow movement. Slots in inlet are made horizontally along the height of pipeline and are directed toward liquid flow. Depth of slots changes from small near pipeline walls to largest near pipeline axis. Opposite to inlet in sampling tube there made is a vertical slot.
EFFECT: increasing sample uniformity and improving accuracy of sample composition determination.
FIELD: measurement equipment.
SUBSTANCE: method involves analysis of an image of mixture surface and determination of coefficient of its non-homogeneity. The investigated mixture is uniformly distributed on a smooth surface and divided into necessary number of portions; digital images of their surfaces with build-up of brightness histograms are obtained. Then, each portion is divided into equal number of parts (probes) with build-up of their brightness histograms. Mixture non-homogeneity coefficient is calculated by comparing digital images of parts (probes) of a piston with an image of the whole portion of the investigated mixture as per brightness histograms.
EFFECT: reducing labour intensity, increasing speed and accuracy of determination of quality of mixture of components that differ as to colour.
SUBSTANCE: sampling device comprises a probe with a sharp-edged cutting edge in the lower part, a flange with a chamfer in the upper part, which is attached coaxially to the rod of a smaller diameter with a measuring scale, and a sample extractor. The cylindrical probe in the lower part is provided with two oval cutting bits with sharp-edged cutting edges. The flange in the upper part with the manufacturing hole made perpendicular to the longitudinal axis of the sampling device is attached to the bar consisting of rods of circular section which are screwed in one another with dimensional manufacturing holes. The openings are made on the surface of the rod in every 500 mm from the oval cutting bits of the probe, and their diameter is equal to diameter of the probe extractor - a rod with a conical groove. Moreover, between the cutting bits and the flange on the probe surface two longitudinal grooves are arranged opposite each other.
EFFECT: reduction of the labour intensity of sampling by providing easier penetration of the sampling device in the estimated mass, providing the possibility of batch sampling from a depth of 1,5-2 m, ensuring easy removal of the sampling device at deep layers of silage.
SUBSTANCE: invention relates to ophthalmology. The cornea storage solution contains a mixture of biological media Ml99 and DMEM, chondroitin sulphate, L-alanyl-L-glutamine, 4-(2-hydroxyethyl)-1-piperazine ethane sulphonic acid (HEPES), sodium pyruvate, sodium β-hydroxybutyrate, gentamicin sulphate, amphotericin B, 2,3,5,7,8-pentahydroxy-6-ethyl naphthalenedione-1,4 (echinochrome A) and hydroxyethyl starch, with the following ratio of components, wt %: medium M199 0.64; medium DMEM 0.447; hydroxyethyl starch 5-10; chondroitin sulphate 2.5; echinochrome A 0.001-0.01; L-alanyl-L-glutamine 0.05425; buffer HEPES 0.5958; sodium pyruvate 0.0055; sodium β-hydroxybutyrate 0.151; gentamicin sulphate 0.01; amphotericin B 0.0000250; distilled water - up to 100.
EFFECT: invention increases storage life of cornea.
1 dwg, 1 tbl, 1 ex
SUBSTANCE: invention relates to the field of cryobiology, cell biology, marine biotechnology and hydrobiology. Processing of marine microalgae cells is carried out with cryoprotective mixture containing penetrating and non-penetrating cryoprotectant and a nutrient medium. Freezing is carried out, followed by preservation in liquid nitrogen. Sterile seawater is taken as the nutrient medium in the following ratio of components, wt %: penetrating cryoprotectant 5-7.5%, non-penetrating cryoprotectant 1.5-3%, sterile sea water - the rest. Prior to freezing, the cells of marine microalgae are incubated in the ice bath for at least 10 minutes. Freezing is carried out in three phases: first, cooled to -25°C at 0.9-1.1 °C/min.; then cooled to -75°C at 2-2.5 °C/min.; then cooled to -196°C, placing marine microalgae cells in liquid nitrogen. Dimethyl sulfoxide (DMSO) or ethylene glycol can be used as penetrating cryoprotectant. Trehalose or polyvinylpyrrolidone (PVP) can be used as non-penetrating cryoprotectant.
EFFECT: invention enables to increase the yield of viable, functionally active cells of microalgae after freezing-thawing and to restore their culture for a week after cryopreservation, while providing a quantitative assessment of the functional state of the microalgae.
3 cl, 1 dwg, 3 tbl, 5 ex
SUBSTANCE: invention relates to medicine. Performed are: intake of donor blood, mixing with preservative glucir in ratio 4^1, separation of donor blood into plasma and erythrocyte mass, with further storage of erythrocyte mass within 30 days at t=+4°C. Hemopreservative 3-oxy-6-methyl-2-ethylpyridine malate is additionally introduced into composition.
EFFECT: invention makes it possible to increase efficiency of protection of erythrocyte cell membranes, limit process of hemolysis, prevent formation of microclots at late terms of erythrocyte mass storage.
2 cl, 6 tbl, 5 ex
SUBSTANCE: invention relates to field of medicine, namely to method of obtaining mineralised bone matrix. Method of obtaining mineralised bone matrix includes mechanical cleaning spongy bone from cartilage tissue and perforation of bone with pin. After that, bone is successively processed with solution of sodium chloride and twin-100 after mixing, with hydrogen peroxide solution, chloroform-ethanol mixture, with volume of all used solutions constituting not less than 5 volumes of spongy bone; after that, it is processed with alcohol-ether mixture, and mixed, all processes of mixing being carried out on magnetic mixer, then, centrifuged and aired, after which bone fragments are frozen, freeze-dried and sterilised under specified conditions.
EFFECT: claimed method makes it possible to reduce time of claimed method realisation, possible storage time of product obtained by claimed method is 3 years, with preservation by product of sterility and biological properties.
SUBSTANCE: invention relates to medicine. Device for preservation of hepatic transplant (1) in normothermia conditions is claimed. Device contains tank (2), in which hepatic transplant (1) is placed in preservation solution, arterial canal (3) of perfusion, portal canal (5) of perfusion, at least, one sensor (7) of consumption in perfusion canal and, at least, one sensor (8) of pressure in perfusion canal. Device additionally contains arterial oxygenator (11), connected with arterial canal (3) of perfusion, portal oxygenator (12), connected with portal canal (5) of perfusion, at least, one arterial perfusion pump (9), at least, one portal perfusion pump (10), module (13) of heat exchange, made with possibility of supporting temperature supplying tank (2) in normothermia condition, and device (14) of pressure and steam control.
EFFECT: invention makes it possible to increase term of hepatic transplant storage.
8 cl, 1 dwg
SUBSTANCE: invention relates to medicine. Anatomic preparation after tissue fixation and preparation is immersed in hexane for 4 weeks, cooled to -18°C, then moved into exiccator, filled with siloxane composition, which contains synthetic caoutchouc, goldhydrochloric acid and hydrosiloxane oligomer, 25 wt % of which is diluted with 75 wt % of hexane. Vacuum of 5 mm Hg is created in it, preparation is cooled to temperature -15°C for 6-8 weeks. After that, preparation is vulcanised in thermostat at temperature 38°C for 4 hours.
EFFECT: invention makes it possible to realise said purpose and simplify the process.
2 dwg, 1 ex
SUBSTANCE: invention may be used for diagnosing telencephalic glyosis in children with congenital infections. The technique involves the morphological examination of cerebral tissue. The white matter tissue fragments are taken from periventricular brain regions at initial postcornu and at interventricular foramen; the histological sections are stained by immunohistochemical staining method using Glial Fibrillary Acidic Protein monoclonal antibodies; reactive astrocytes are detected, and the presence thereof by more than 30%, telencephalic glyosis is diagnosed.
EFFECT: use of the declared technique enables the most accurate diagnosis of telencephalic glyosis in congenital infections in children, and promotes making an early, accurate instrumental diagnosis.
SUBSTANCE: invention relates to agriculture. The following steps are performed: (a) providing a composition comprising or consisting of a carrier and one or several repellent compounds selected from para-cymene, gamma-terpinene, alpha-terpinene, alpha-phellandrene and/or epicyngibberene, (b) adding of the said composition, one or several times to a plurality of cultivated plants. The said composition is applied for scaring whiteflies. To attract whiteflies a composition is used which comprises a carrier and at least one compound as an active ingredient, selected from the group consisting of beta-phellandrene, limonene and\or 2-carene.
EFFECT: invention enables to improve the efficiency of whitefly control.
18 cl, 3 dwg, 1 tbl, 5 ex
SUBSTANCE: invention relates to agriculture. The agent against crop fungal diseases Bipolaris sorokiniana, Fusarium oxsporium and Aspergillus niger contains an active substance from the dithiazinane family - α,ω-bis-( 1,3,5-dithiazinan-5-yl)-3-oxopantane of formula in form of a 0.1-0.15% solution in dimethylformamide.
EFFECT: invention enables to use a readily available agent obtained using a simple technique.
SUBSTANCE: invention relates to medicine. Before washing and submerging liver into preserving solution Costodiol, perfluorane, non-enriched with oxygen, is added to it in ratio 4:1 respectively. Under conditions of cold hypothermia preservation is performed for 8 hours at temperature +5°C.
EFFECT: invention makes it possible to increase efficiency of anti-ischemic protection of liver transplant.
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