CN110003951B - Paraffin for pathological semi-thin section and preparation method thereof - Google Patents
Paraffin for pathological semi-thin section and preparation method thereof Download PDFInfo
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- CN110003951B CN110003951B CN201910375676.1A CN201910375676A CN110003951B CN 110003951 B CN110003951 B CN 110003951B CN 201910375676 A CN201910375676 A CN 201910375676A CN 110003951 B CN110003951 B CN 110003951B
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- C10—PETROLEUM, GAS OR COKE INDUSTRIES; TECHNICAL GASES CONTAINING CARBON MONOXIDE; FUELS; LUBRICANTS; PEAT
- C10G—CRACKING HYDROCARBON OILS; PRODUCTION OF LIQUID HYDROCARBON MIXTURES, e.g. BY DESTRUCTIVE HYDROGENATION, OLIGOMERISATION, POLYMERISATION; RECOVERY OF HYDROCARBON OILS FROM OIL-SHALE, OIL-SAND, OR GASES; REFINING MIXTURES MAINLY CONSISTING OF HYDROCARBONS; REFORMING OF NAPHTHA; MINERAL WAXES
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- C10G73/38—Chemical modification of petroleum
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Abstract
The invention belongs to the technical field of pathology and discloses novel paraffin for pathological semi-thin sections and a preparation method thereof, wherein the paraffin is heated and boiled (100 ℃), tetramethylethylenediamine with the volume proportion of 1% and ammonium persulfate with the volume proportion of 1% are added into liquid paraffin, and the liquid paraffin is uniformly mixed in a vortex oscillator or a stirrer, so that the novel paraffin is prepared. The improved novel paraffin wax has better ductility and proper viscosity. The novel paraffin wax can be used for manufacturing pathological semi-thin sections, can remarkably reduce the technical difficulty of manufacturing the semi-thin sections and is beneficial to improving the pathological technical level of most primary hospitals.
Description
Technical Field
The invention belongs to the technical field of pathology, and particularly relates to novel paraffin for pathological semi-thin sections and a preparation method thereof.
Background
Currently, the closest prior art includes ultra-thin sections and resin semi-thin sections for electron microscopy applications. The former preparation apparatus requires an ultra-thin microtome, uses a glass knife or a diamond knife, and an embedding medium for embedding a specimen must have a certain hardness, and is usually embedded by resin polymerization, in order to cut the specimen into thin sections. The latter is a technique of slicing a resin-embedded sample into slices having a thickness of 0.5 to 2 μm by an ultra-thin microtome. Since the slice thickness obtained by this technique is generally between that of an ultrathin slice and that of a paraffin slice, it is called a semi-thin slice and is usually used for tissue localization before ultrathin slicing. The two methods are expensive, complex in operation, time-consuming, labor-consuming and dependent on expensive special equipment (including an electron microscope, a half-microtome and the like), so that the method is mainly used for scientific research, cannot meet the requirements of clinical pathological practices of primary hospitals, and has not been popularized and applied in clinical pathological diagnosis work.
Paraffin sectioning is the most widely used method in conventional histological sectioning techniques. The paraffin section is not only used for observing the morphological structure of normal cell tissues, but also is a main method for researching, observing and judging the morphological change of the cell tissues in the subjects of pathology, legal medicine and the like. In pathological diagnosis, particularly in diagnosis of malignant tumors such as lymphoma, it is required that the thinner the slice thickness, the better the slice thickness is, in order to distinguish nuclear fission images in nuclei. In clinical practice, only 4-micron-thick sections can be cut, and due to the limitation of viscosity and ductility of paraffin, the aim of cutting thinner sections (such as 1-2 microns) cannot be achieved in clinical practice.
In summary, the problems of the prior art are as follows:
in clinical practice, only a slice with a thickness of 4 microns can be cut, and due to the limitation of viscosity and ductility of paraffin, the aim of cutting a thinner slice (1-2 microns), namely preparing a paraffin semi-thin slice, cannot be achieved in clinical practice.
The paraffin semi-thin section is prepared on the premise of not depending on the operation technical level of clinical pathology technicians and comprehensively considering instruments and equipment of most primary hospitals.
The difficulty of solving the technical problems is as follows:
improving the viscosity and ductility of paraffin is the key to making semi-thin sections.
The improved method is simple and easy to implement, low in cost and good in effect.
The significance of solving the technical problems is as follows:
the hospital pathology technical room is an important platform for developing medical and health services in daily life in hospitals, has a very close relation with the disease diagnosis of patients, and meanwhile, the high-quality pathology technical work can directly have a positive influence on the daily economic benefit, the internal structure construction and the long-term development in the future of the hospitals. The resolution of the paraffin semi-thin section on the fine tissue structure, the cell content, the cell arrangement level and the like under the disease state is better than that of the conventional paraffin section. The invention can make a semi-thin section with the thickness of 1 micron by using the novel paraffin embedded tissue, can simply and easily improve the quality of pathological sections and meet the requirements of pathological work of most primary hospitals.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides novel paraffin for pathological semi-thin sections and a preparation method thereof.
The invention is realized in such a way, and the novel paraffin is used for pathological semi-thin sections. The novel paraffin preparation method for pathological semi-thin section comprises the following steps:
firstly, heating and boiling paraffin (100 ℃);
secondly, adding 1 volume percent of tetramethylethylenediamine and 1 volume percent of ammonium persulfate into the liquid paraffin;
thirdly, evenly mixing the liquid paraffin in a vortex oscillator or a stirrer;
and fourthly, preparing the novel paraffin.
The Tetramethylethylenediamine (TEMED) is named as: n, N, N ', N' -tetramethylethylenediamine with the molecular formula: (CH3)2NCH2CH2N (CH3) 2).
Further, in the present invention, the volume ratio of tetramethylethylenediamine to ammonium persulfate may be adjusted, but is not preferably higher than 5%.
The invention also aims to provide the novel paraffin for pathological semi-thin sections, which is prepared by the preparation method of the novel paraffin for pathological semi-thin sections.
In summary, the advantages and positive effects of the invention are:
the invention relates to an improved paraffin wax formula, and the improved novel paraffin wax has better ductility and proper viscosity. The novel paraffin wax can be used for manufacturing pathological semi-thin sections, can remarkably reduce the technical difficulty of manufacturing the semi-thin sections and is beneficial to improving the pathological technical level of most primary hospitals. The specific effects are summarized as follows:
compared with the conventional paraffin section, the invention has better resolution in the aspects of observing the tissue fine tissue structure, the cell content, the cell arrangement level and the like; compared with the traditional methods of embedding semi-thin slices by resin and the like, the method has the advantages that the operation method is simpler and easier, and the cost is low because the method does not depend on an ultrathin slicer and a special diamond blade; compared with the electron microscope ultrathin section, the invention does not need to rely on expensive equipment such as an electron microscope and the like for observation, thereby being more suitable for popularization in the pathology department of the primary hospitals.
The invention provides a brand-new preparation method of a semi-thin paraffin slice, and fills up the technical blank at home and abroad.
Drawings
Fig. 1 is a flow chart of a method for preparing novel paraffin for pathological ultrathin section provided by the embodiment of the invention.
FIG. 2 is a comparison graph of the staining effect of normal paraffin sections and novel paraffin sections on normal tissues, provided by the embodiment of the invention. FIG. 2A Normal tonsil tissue Normal Paraffin-Embedded section HE staining (4 micron) B Normal tonsil tissue novel Paraffin-Embedded section HE staining (4 micron) C Normal tonsil tissue novel Paraffin-Embedded section HE staining (1 micron) D Normal gallbladder wall tissue Normal Paraffin-Embedded section HE staining (4 micron) E Normal gallbladder wall tissue novel Paraffin-Embedded section HE staining (4 micron) F Normal gallbladder wall tissue novel Paraffin-Embedded section HE staining (1 micron) G Normal mammary gland connective tissue Normal Paraffin-Embedded section HE staining (4 micron) H Normal mammary gland connective tissue novel Paraffin-Embedded section HE staining (4 micron) I Normal mammary gland connective tissue novel Paraffin-Embedded section HE staining (1 micron)
FIG. 3 is a comparison graph of the staining effect of normal paraffin section and new paraffin section on tumor tissue. HE staining (4 micron) of lung cancer common paraffin embedded section, B novel paraffin embedded section, HE staining (4 micron) of lung cancer, C novel paraffin embedded section, HE staining (1 micron) of lung cancer, D liver cancer common paraffin embedded section, HE staining (4 micron) of liver cancer, E novel paraffin embedded section, HE staining (4 micron) of liver cancer, F novel paraffin embedded section, HE staining (1 micron) of G lymphoma common paraffin embedded section, HE staining (4 micron) of H lymphoma novel paraffin embedded section, HE staining (4 micron) of lymphoma I novel paraffin embedded section, 1 micron)
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In clinical practice, only 4-micron-thick sections can be cut, and due to the limitation of viscosity and ductility of paraffin, the aim of cutting thinner sections (such as 1-2 microns) cannot be achieved in clinical practice.
To solve the above problems, the following detailed description will be made of the application of the present invention with reference to the accompanying drawings.
As shown in fig. 1, the preparation method of the novel paraffin for pathological semi-thin section provided by the embodiment of the invention comprises the following steps:
s101: the paraffin was heated to boiling (100 ℃).
S102: adding 1 volume percent of tetramethylethylenediamine and 1 volume percent of ammonium persulfate into the liquid paraffin.
S103: the liquid paraffin was mixed well in a vortex shaker or stirrer.
S104: and preparing the novel paraffin.
The Tetramethylethylenediamine (TEMED) has the Chinese name: n, N, N ', N' -tetramethylethylenediamine with the molecular formula: (CH3)2NCH2CH2N (CH3) 2).
In the present invention, the volume ratio of tetramethylethylenediamine to ammonium persulfate may be adjusted, but is not preferably higher than 5%.
The effectiveness of the method provided by the invention is verified by combining the attached drawings.
And melting the prepared novel paraffin in a paraffin embedding machine, and carrying out paraffin embedding and slicing according to conventional operation. At this time, the viscosity and ductility of the paraffin section are improved, and the continuous section is not easy to break and wrinkle, so that a semi-thin section with the thickness of 1 micron can be obtained.
Fig. 2 and fig. 3 are graphs comparing the staining effect of normal tissues and tumors by normal paraffin sections and novel paraffin sections provided by the embodiment of the invention. The novel paraffin can be used for preparing a semi-thin section with the thickness of 1 micron according to the clinical pathological diagnosis requirement, and can also be used for cutting a common section with the thickness of 4 microns.
As shown in fig. 2, it can be seen from comparison that the slice thickness obtained by normal paraffin cutting is thick, HE staining (hematoxylin-eosin staining) background is deep, and nucleus staining is deep, so that structures such as nucleus division and the like cannot be distinguished. The slice cut by the improved novel paraffin is thinner, the staining background is shallow, the cell nucleus is clearly stained, and the nuclear fission image can be clearly distinguished. The novel paraffin can reduce the technical difficulty of manufacturing the semi-thin section, and is favorable for improving the pathological technical level of most primary hospitals.
FIG. 2A staining of normal tonsil tissue normal paraffin-embedded section HE (4 microns), staining of normal tonsil tissue novel paraffin-embedded section HE (1 micron), staining of normal gallbladder wall tissue normal paraffin-embedded section HE (4 microns), staining of normal gallbladder wall tissue novel paraffin-embedded section HE (1 micron), staining of normal mammary gland connective tissue normal paraffin-embedded section HE (4 microns), staining of normal mammary gland connective tissue novel paraffin-embedded section HE (4 microns), staining of normal mammary gland connective tissue normal paraffin-embedded section HE (1 micron).
FIG. 3A staining of a lung cancer common paraffin-embedded section HE (4 microns), staining of a novel paraffin-embedded section HE of a lung cancer B (4 microns), staining of a novel paraffin-embedded section HE of a lung cancer C (1 micron), staining of a liver cancer D (4 microns), staining of a liver cancer E (4 microns), staining of a liver cancer F (1 micron), staining of a liver cancer G (4 microns), staining of a lymphoma G (4 microns), staining of a liver cancer H (4 microns), and staining of a lymphoma I (1 micron).
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (2)
1. A preparation method of paraffin for pathological semi-thin sections is characterized by comprising the following steps:
firstly, heating and boiling paraffin;
secondly, adding 1 volume percent of tetramethylethylenediamine and 1 volume percent of ammonium persulfate into the liquid paraffin;
thirdly, evenly mixing the liquid paraffin in a vortex oscillator or a stirrer;
and fourthly, preparing the paraffin.
2. A paraffin for pathological semi-thin section prepared by the method for preparing paraffin for pathological semi-thin section as claimed in claim 1.
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| CN201910375676.1A CN110003951B (en) | 2019-05-07 | 2019-05-07 | Paraffin for pathological semi-thin section and preparation method thereof |
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| CN201910375676.1A CN110003951B (en) | 2019-05-07 | 2019-05-07 | Paraffin for pathological semi-thin section and preparation method thereof |
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