RU2104513C1 - Method of histologic specimen preparation - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: fixation, dehydration, clarification and paraffinization are performed in domestic microwave oven of power equal to 450 W. Fixation is carried out for 20 s at 90% power. After draining the fixing the liquid and pouring over the piece of tissues with 96% spirit, it is placed in microwave oven for 10 min at 20-30% power with additional water load of total volume of 1000 ml from four sides of vessel containing the piece of tissues. Dehydration is performed three times in similar mode of oven operation. This done, piece of tissue is poured over with mixture of spirit with chloroform in relation of 1:1 and placed in microwave oven for 5 min at 30% power and total water load of 1250 ml positioned over circumference of vessel with piece of tissue and optimal radius of 3 cm in five vessels. After pouring over with clean chloroform, vessel with piece of tissue is placed in microwave oven again for 10 min at 70 per cent power. Paraffinization is performed by pouring over the piece of tissue with mixture of chloroform and paraffin in traditional way. Piece of tissue is placed in microwave oven for 5 min at 50% power and previous water load. Then it is poured over with liquid paraffin in traditional way and placed in microwave oven again for 15 min at 40% power and the same water load as in previous step. EFFECT: reduction of histology specimen preparation time from 10-12 days to 1 h 20 min with high quality of specimen preserved.

Description

 The invention relates to medicine, more specifically, to a technique for the manufacture of histological preparations of tissue samples, and can be used in differential and clarifying diagnostics of various pathological conditions of the body.

 Known methods for preparing tissue samples for microscopic analysis, including their fixation, dehydration, waxing, after which a histological specimen for microscopic examination is prepared from microscope specimens from tissue samples (G.A. Merkulov, Pathology and histological technology course, Medgiz, 1956). The listed stages of the preparation of histological samples are performed manually by moving them to various solutions.

 The production of histological samples according to the known method proposed by G.A. Merkulov allows one to obtain high-quality samples while observing all technological methods, namely, multiple transfer to various solutions for a long time, sometimes up to 48 hours, washing in running water for several hours . All this leads to the fact that the total production time of histological samples increases and reaches 7-8 days. Only after this time is it possible to prepare histological sections of tissues, their interpretation, final differential diagnosis.

 Thus, special treatment for the patient can be prescribed 10-12 days after the discovery of any pathology. Such a delay in special treatment aggravates the patient's condition, and in the presence of a malignant disease is fraught with significant progression of the process and complications in its further treatment.

 There is an accelerated method for the manufacture of histological preparations with the so-called cito-diagnostics, when a piece of tissue taken for histological examination is frozen with liquid nitrogen on a cryostat. Then a histological preparation is prepared, a study of the latter is carried out, and after 10-50 minutes after taking a piece of tissue, the nature of the patient’s disease is determined (G. Merkulov. Pathology and histological technique. - L .: Medicine, 1969).

 However, such an accelerated method for the manufacture of histological samples cannot be used to study all tissues of a living organism. Rapid freezing of mucous membranes, i.e. tissues having a loose structure, leads to their change, and not only the appearance, but also the structure of the cells, which is especially important for establishing the correct diagnosis. As for fatty and flabby tissues, they generally do not lend themselves to rapid freezing. In addition, the method of accelerated preparation of histological preparations cannot be applied for the study of bone tissue. These negative properties force in the future to return to the re-examination of histological preparations of the same tissues, but made according to the known long-term method.

 The aim of the invention is to reduce the manufacturing time of histological preparations of various tissues with high quality.

 The goal is achieved by the fact that a piece of tissue taken for the manufacture of a histological preparation is immersed in a container with a 10% formalin solution and placed in a household microwave (MV) oven with a total power of 450 W for 20 s with a power mode of 90%. Be sure to use a water load - 250 ml of water in a jar. Next, the container is removed from the furnace, the fixing liquid is drained, after which the piece is filled with 96% alcohol and placed in the MV-furnace for 10 minutes at a capacity of 20-30% with an additional water load in a total volume of 1000 ml on four sides of the container with a piece. Filling with alcohol (dehydration) at the same power mode of the MV-furnace three times. Before waxing, a piece of tissue is poured with a mixture of 96% alcohol with chloroform in a ratio of 1: 1, placed again in the MV oven for 5 minutes at a power of 30% and a total water load of 1250 ml, located around a circle around a container with a piece of fabric with an optimal radius 3 cm in five containers. Then the container with a slice is poured with pure chloroform and again placed in the MV-oven for 10 min at a power of 70%. The water load is the same as in the previous step. At the last stage (paraffinization), a piece of tissue is poured with a mixture of chloroform and paraffin according to the traditional method and placed in a MV-oven for 5 min at a power of 50% with a previous water load. After that, the piece is poured with pure liquid paraffin according to the traditional method and placed in the MV-oven for 15 minutes at a power of 40% with the water load of the previous stage.

 A comparative analysis of the developed method for the manufacture of histological preparations with the known one shows that it has an inventive step and is new, since for a specialist it does not follow explicitly from the level of modern development of this branch of medicine.

 The level of development of the technology for the manufacture of histological preparations in the available scientific and medical literature does not have information about the use of household microwave ovens for the manufacture of samples of various tissues. This circumstance allows us to consider the proposed method significantly different from the currently known.

 The developed method is industrially applicable, as it can be used in wide medical practice of pathomorphological laboratories of medical institutions.

 The proposed method was used for the preparation of histological samples of various tumor tissues - breast, lymph nodes, uterus, ovaries, lung, soft tissues. In the process of developing the method, more than 400 histological preparations of various types of tissues were manufactured. In all cases, the quality of the preparations was high, which made it possible to easily interpret them and carry out differential diagnosis of tumor formations of different localization.

 The technical result of the use of the developed method consists in the possibility of significantly reducing the time of manufacture of histopathological preparations, obtaining their high quality, increasing the number of manufactured preparations, not requiring additional chemicals, the cost of acquiring a household microwave oven pays off in 1.5-2 months.

Claims (1)

 A method of manufacturing histological preparations, including tissue fixation, dehydration, enlightenment and waxing, characterized in that, in order to reduce the manufacturing time of histological preparations of various tissues, fixation, dehydration, enlightenment, waxing is carried out in a household microwave (MV) oven, while fixing is carried out for 20 s at a power of 90% (from 450 W), after draining the fixing liquid and pouring a piece of 96% alcohol, it is placed in the MV-furnace for 10 min at a power of 20 30% with an additional water load of with a volume of 1000 ml on four sides of the container with a piece of tissue, dehydration is carried out three times in the same mode of the MV oven, then a piece of tissue is poured with a mixture of 96% alcohol with chloroform in a ratio of 1 1, placed in the MV oven for 5 min at power 30% and a total water load of 1250 ml, located around the circumference of the container with a piece with an optimal radius of 3 cm in five containers, after pouring with pure chloroform, the container with a piece of fabric is again placed in the MV oven for 10 min at a power of 70% and waxing is carried out by pouring pieces of fabric with a mixture of chloe the form and paraffin according to the traditional method, put it in the MV-furnace for 5 min at a power of 50% with a previous water load, then fill it with liquid paraffin according to the traditional method and again put in the MV-furnace for 15 min at a power of 40% and the water load of the previous stage.