CN102183387B - Benzene-free environment-friendly tissue or slice clearing liquid - Google Patents
Benzene-free environment-friendly tissue or slice clearing liquid Download PDFInfo
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- CN102183387B CN102183387B CN 201110023746 CN201110023746A CN102183387B CN 102183387 B CN102183387 B CN 102183387B CN 201110023746 CN201110023746 CN 201110023746 CN 201110023746 A CN201110023746 A CN 201110023746A CN 102183387 B CN102183387 B CN 102183387B
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Abstract
The invention provides a benzene-free tissue or slice clearing liquid which comprises, by volume, the following components: 550-880 parts of food-grade vegetable oil extractants, 50-440 parts of turpentine, 0-50 parts of acetone, and 10-20 parts of limonene. The benzene-free tissue or slice clearing liquid adopts a composite component formula; all the components are chemical reagents with nontoxicity and innocuousness; and all the components perform the clearing effect together on cells and tissues. By cell morphology experiment, immunohistochemistry experiment, molecular pathology experiment and nucleic acid extraction experiment, it is proved that the clearing liquid can completely substitute xylene and is applicable to related pathology experiment.
Description
Technical field
The present invention relates to external diagnosis reagent, be specifically related to a kind of benzene-free type environmental protection organization or cut into slices transparent liquid and application thereof, belong to the pathological experiment technical field.
Background technology
The history in existing more than 150 year of histopathologic birth in all hospitals, as long as operation is arranged, is made histopathologic diagnosis by hospital pathology department to the operation tissue samples.
Routine paraffin wax embedding HE stained preparation is current basic fundamental in order to handling tissue samples, and it is the basis that pathological diagnosis method is learned, and also is the evaluation reference standard of many new methods, new technology quality.Traditional method is that tissue samples is placed in the cylinder for the treatment of fluid, pours successively or call in different disposal liquid into, and sample soaks in accordance with regulations successively different time and finishes processing section, dyeing, mounting etc. in each reagent, and whole process needs more than 30 step.Whole process is used formaldehyde, dimethylbenzene, mercury oxide, the toxic chemical reagent of ammoniacal liquor in a large number, and the pathology technique personnel place oneself in the midst of toxic environment for a long time, and the reagent emission of serious pollution of environment.Because the tolerance of agents useful for same is poor, complicated operation, reagent consumption is large, pathology film-making high-quality sheet low (whole nation statistics is 40～70%), impact diagnosis.The large absolutely hospital of counting joins reagent voluntarily simultaneously, lacks unified standard.Therefore the standardized production of pathology industry organization sample disposal liquid articles for use, without poison, the socialization supply and demand, be the inexorable trend of industry development.
Use benzene,toluene,xylene to be used for the transparent and paraffin embedding of human body and animal vegetable tissue sample, carry out the existing history more than 160 years of Human pathology and animals and plants pathological research.Benzene, toluene and dimethylbenzene are that the poisonous and harmful chemicals of generally acknowledging has potential carcinogenicity.Especially dimethylbenzene is a kind of traditional phenyl ring aromatic hydrocarbon solvent that very is widely used in pathology film-making and the dyeing, and the World Health Organization (WHO) has declared that it is to the toxicity of central nervous system.Test and clinical research show that all sucking xylene steam causes central nervous system (CNS) depressive symptom, and such as headache, dizzy, nausea and vomiting, dimethylbenzene is positive through micronucleus test, and human body is also had carcinogenesis.Dimethylbenzene is injury doctor's arch-criminal, allows the arch-criminal of the thrilling flavor of whole pathology department.At present the substitute of existing in addition existing part dimethylbenzene is used for the transparent and paraffin-embedded experimental study of human body and animal vegetable tissue sample, for example application number be 200810136961.X Patent Application Publication a kind of routine paraffin wax flaking HE dyeing non-xylol whole-course treatment liquid for biological organization sample.Yet this murder by poisoning reagent formaldehyde that has still adopted serious environment pollution based on the fixing treating fluid of tradition tissue.For example also have terebinthina and the alternative simple dimethylbenzene of dimethylbenzene mixed liquor are carried out the research of transparency of organization, existing discussion stearic acid paraffin mixed liquor and dimethylbenzene are as the difference of transparency of organization agent, the report that perhaps uses isopropyl alcohol to estimate in the transparency of organization process in addition.Yet these prior aries still can't be abandoned the use of dimethylbenzene fully, perhaps can't reach preferably transparency of organization effect.
Therefore, the alternative dimethylbenzene of the substitute of research and development dimethylbenzene has great importance for pathological experiment.
Summary of the invention
The object of the present invention is to provide a kind of benzene-free type tissue or the transparent liquid of cutting into slices, can satisfy routine pathology diagnosis, immunohistochemical diagnosis technology, diagnosis of molecular biology technical requirement, the tissue effect that processes is better than conventional art, to substitute the pathological experiment that dimethylbenzene is applied to be correlated with fully.The concentration of each reagent of the present invention is mass percentage concentration.
In order to reach purpose of the present invention, adopt following technical scheme:
The invention provides a kind of benzene-free type tissue or the transparent liquid of cutting into slices, it comprises the component of following volume parts:
Food-grade vegetable oil extractant 550-880 part, terebinthina 50-440 part, acetone 0-50 part, hesperidene 10-20 part.
In the preferred embodiment of the present invention, described benzene-free type tissue or the transparent liquid of cutting into slices comprise the component of following volume parts: 660 parts of food-grade vegetable oil extractants, 300 parts in terebinthina, 25 parts in acetone, 15 parts of hesperidenes.
The instant grade solvent oil of wherein said food-grade vegetable oil extractant for the vegetable fat extractant belongs to hydrocarbon mixture, the model such as can adopt No. 80, No. 100, is preferably food-grade vegetable oil extractant No. 100.
Wherein said terebinthina is preferably the tasteless terebinthina of refining.
Wherein said benzene-free type tissue or the transparent liquid of cutting into slices can also comprise surfactant, for example quaternary ammonium salt surface active agent or fluorine class surfactant, and the volume parts of adding is 40-60 part, its concentration is 0.5-2%, preferred 1%.
Described quaternary ammonium salt surface active agent is to be selected from octyl group-decyl-alkyl dimethyl ammonium chloride, Quaternium 24, DDAC, didecyl dimethyl ammonium bromide, preferred octyl group-decyl-alkyl dimethyl ammonium chloride.
The preparation of benzene-free type tissue provided by the invention or the transparent liquid of cutting into slices is that each component is got final product in described ratio mixing at normal temperatures.
Benzene-free type tissue provided by the invention or cut into slices transparent liquid when using can not revised traditional grade dehydration of alcohol and dimethylbenzene transparent process, and directly substitutes waxdip after the dimethylbenzene transparency of organization.In paraffin method, except hyaline tissue's sample material, also can be used as the solvent of dissolving paraffin and sealing natural gum in addition.Be specially adapted to dewaterer.The normal dyeing method be can not only be used for, special staining and tissue chemical technology also are applicable to.
Benzene-free type tissue provided by the invention or the transparent liquid of cutting into slices adopt the composite parts prescription, and its whole components are nontoxic chemical reagent, the common transparent effect of carrying out cell and tissue of each component.Experiment showed, fully through cytomorphology experiment, immunohistochemical assay, molecular pathology experiment and nucleic acid extraction and can substitute the pathological experiment that dimethylbenzene is applied to be correlated with.
Description of drawings
Fig. 1 is the human breast carcinoma tissue figure (200 times of LEICA DM4000B cinephotomicrographies) that adopts benzene-free type tissue of the present invention or the transparent liquid of cutting into slices is processed.
Fig. 2 is the people's renal tissue after stain chromatic graph (200 times of LEICA DM4000B cinephotomicrographies) that adopts benzene-free type tissue of the present invention or the transparent liquid of cutting into slices is processed.
Embodiment
The invention will be further described below in conjunction with implementing, and it should be understood that these embodiment only are used for the purpose of illustration, never limit protection scope of the present invention.
No. 80, No. 100 food-grade vegetable oil extractants: No. 80, No. 100 solvent oils of instant grade, for the vegetable fat extractant belongs to hydrocarbon mixture, originate from Dalian.
Terebinthina: for the tasteless terebinthina of refining, originate from Guangxi, CAS number: 8006-64-2.
Hesperidene: originate from the U.S., CAS:5989-27-5.
Embodiment 1
No. 100 food-grade vegetable oil extractant---------880ml
Terebinthina----------------------------------50ml
Acetone------------------------------------50ml
Hesperidene------------------------------------20ml
Said components is miscible and get final product at normal temperatures.
Embodiment 2
No. 100 food-grade vegetable oil extractant--------550ml
Terebinthina-------------------------440ml
Hesperidene----------------------------10ml
Said components is miscible and get final product at normal temperatures.
Embodiment 3
No. 80 food-grade vegetable oil extractant----------660ml
Terebinthina--------------------------300ml
Acetone-----------------------------25ml
Hesperidene-----------------------------15ml
Said components is miscible and get final product at normal temperatures.
Embodiment 4
No. 80 food-grade vegetable oil extractant---------730ml
Terebinthina-------------------------200ml
Acetone----------------------------10ml
Hesperidene----------------------------20ml
Octyl group-decyl of 1%-alkyl dimethyl ammonium chloride----40ml
Said components is miscible and get final product at normal temperatures.
Test effect
Experimental results show that, adopt the benzene-free type tissue of embodiment 1-4 or the human breast carcinoma tissue of the transparent liquid processing of cutting into slices, see Fig. 1 (200 times of LEICA DM4000B cinephotomicrographies), C-erbB-2 gene expression protein immunization histochemical stain antigen location reticulates structure with cell membrane, and the nucleus counterstain is clear.
Adopt the benzene-free type tissue of embodiment 1-4 or people's renal biopsy of the transparent liquid processing of cutting into slices, see Fig. 2 (200 times of LEICA DM4000B cinephotomicrographies), glomerular basement membrane PASM dyeing shows good institutional framework and glomerular basement membrane form.
As seen, experimental results show that through cytomorphology experiment, immunohistochemical assay, molecular pathology experiment and nucleic acid extraction, the transparent liquid effect of benzene-free type environmental protection organization of the present invention is soft, be difficult for causing Materials for slide making drawdown deformation, sclerosis embrittlement, can improve the serviceability rate of section, and has comparatively good transparency of organization effect, techtology standard of comparison after the processing is consistent, can not affect that follow-up group spy dyes, the analysis requirement of immunohistochemical staining and molecular pathology, can substitute the pathological experiment that dimethylbenzene is applied to be correlated with fully.
The above only is preferred embodiment of the present invention, only is illustrative for the purpose of the present invention, and nonrestrictive.Those skilled in the art is understood, and can carry out many changes to it in the spirit and scope that claim of the present invention limits, revise, even equivalence, but all will fall within the scope of protection of the present invention.
Claims (8)
1. a benzene-free type tissue or the transparent liquid of cutting into slices, it comprises the component of following volume parts: food-grade vegetable oil extractant 550-880 part, terebinthina 50-440 part, acetone 0-50 part, hesperidene 10-20 part.
2. benzene-free type tissue according to claim 1 or the transparent liquid of cutting into slices, it comprises the component of following volume parts: 660 parts of food-grade vegetable oil extractants, 300 parts in terebinthina, 25 parts in acetone, 15 parts of hesperidenes.
3. benzene-free type tissue according to claim 1 and 2 or the transparent liquid of cutting into slices, wherein said food-grade vegetable oil extractant is No. 80 or No. 100 food-grade vegetable oil extractants.
4. benzene-free type tissue according to claim 1 and 2 or the transparent liquid of cutting into slices, wherein said terebinthina is the tasteless terebinthina of refining.
5. benzene-free type tissue according to claim 1 and 2 or the transparent liquid of cutting into slices wherein also comprise surfactant, are quaternary ammonium salt surface active agent or fluorine class surfactant, and the volume parts of adding is 40-60 part, and its concentration is 0.5-2%.
6. benzene-free type tissue according to claim 5 or the transparent liquid of cutting into slices wherein also comprise surfactant, are quaternary ammonium salt surface active agent or fluorine class surfactant, and the volume parts of adding is 40-60 part, and its concentration is 1%.
7. benzene-free type tissue according to claim 6 or the transparent liquid of cutting into slices, wherein said quaternary ammonium salt surface active agent is to be selected from octyl group-decyl-alkyl dimethyl ammonium chloride, Quaternium 24, DDAC, didecyl dimethyl ammonium bromide.
8. the application of each described benzene-free type tissue of claim 1-7 or the transparent liquid of cutting into slices is to make for Histopathology and animal or plant specimen, is used for routine pathology diagnosis, immunohistochemical diagnosis technology, diagnosis of molecular biology technology.
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| CN102183387B true CN102183387B (en) | 2013-04-10 |
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102507284A (en) * | 2011-11-11 | 2012-06-20 | 福州迈新生物技术开发有限公司 | Tissue paraffin removing liquid |
| CN102854043B (en) * | 2012-09-10 | 2014-08-06 | 韩占强 | Medical slice production solvent |
| CN103471888B (en) * | 2013-09-09 | 2016-02-03 | 四川省中医药科学院 | Pure-natural biological film-making clarifier and the method with this clarifier paraffin wax flaking |
| CN104655460B (en) * | 2013-11-25 | 2017-09-12 | 无锡市江原实业技贸总公司 | A kind of transparent dewaxing liquid of pathological tissue and preparation method thereof |
| CN105823665A (en) * | 2016-05-13 | 2016-08-03 | 北京九州柏林生物科技有限公司 | Biological tissue sample preparation sleeve solution |
| CN106053187A (en) * | 2016-05-20 | 2016-10-26 | 北京九州柏林生物科技有限公司 | Liquid set for dyeing biological tissue sample slices |
| CN109470534B (en) * | 2018-11-12 | 2021-05-25 | 山东骏腾医疗科技有限公司 | Pathological tissue transparent dewaxing liquid and preparation method thereof |
| CN111380735B (en) * | 2018-12-28 | 2022-08-23 | 哈尔滨格林标本技术开发有限公司 | Efficient and environment-friendly biological tissue treatment liquid and method for treating tissue slices by using same |
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| CN1068653A (en) * | 1991-08-26 | 1993-02-03 | 广西农学院 | Tranparency agent for biological sample slide |
| CN1068889A (en) * | 1992-07-11 | 1993-02-10 | 南京大学 | The method and apparatus of quick preparing human tissue pathological section |
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| JP2004069666A (en) * | 2002-08-08 | 2004-03-04 | Tsuerun:Kk | Technique for cutting out and aligning part of paraffin section |
| CN101871855A (en) * | 2009-04-27 | 2010-10-27 | 上海蓝盎电子科技发展有限公司 | Novel pathological section clarifier |
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2011
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Patent Citations (5)
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|---|---|---|---|---|
| CN1068653A (en) * | 1991-08-26 | 1993-02-03 | 广西农学院 | Tranparency agent for biological sample slide |
| CN1068889A (en) * | 1992-07-11 | 1993-02-10 | 南京大学 | The method and apparatus of quick preparing human tissue pathological section |
| CN1337462A (en) * | 2000-08-03 | 2002-02-27 | 中国人民解放军兰州军区兰州总医院 | Parafin structure slice dewaxing and clearing agent |
| JP2004069666A (en) * | 2002-08-08 | 2004-03-04 | Tsuerun:Kk | Technique for cutting out and aligning part of paraffin section |
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