CN110779785A - Novel formaldehyde-free tissue fixing solution - Google Patents
Novel formaldehyde-free tissue fixing solution Download PDFInfo
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- CN110779785A CN110779785A CN201911117669.8A CN201911117669A CN110779785A CN 110779785 A CN110779785 A CN 110779785A CN 201911117669 A CN201911117669 A CN 201911117669A CN 110779785 A CN110779785 A CN 110779785A
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- 239000000834 fixative Substances 0.000 claims abstract description 52
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 claims abstract description 37
- 239000000203 mixture Substances 0.000 claims abstract description 30
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- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 claims abstract description 24
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- 239000005715 Fructose Substances 0.000 claims abstract description 17
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- 238000002360 preparation method Methods 0.000 claims abstract description 10
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- 238000002156 mixing Methods 0.000 claims description 11
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- 241001465754 Metazoa Species 0.000 claims description 8
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 7
- 235000011181 potassium carbonates Nutrition 0.000 claims description 7
- FSVCELGFZIQNCK-UHFFFAOYSA-N N,N-bis(2-hydroxyethyl)glycine Chemical compound OCCN(CCO)CC(O)=O FSVCELGFZIQNCK-UHFFFAOYSA-N 0.000 claims description 5
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 5
- 238000009833 condensation Methods 0.000 claims description 5
- 230000005494 condensation Effects 0.000 claims description 5
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- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 claims description 5
- 229960004889 salicylic acid Drugs 0.000 claims description 5
- 239000001818 polyoxyethylene sorbitan monostearate Substances 0.000 claims description 4
- 235000010989 polyoxyethylene sorbitan monostearate Nutrition 0.000 claims description 4
- 235000015497 potassium bicarbonate Nutrition 0.000 claims description 4
- 239000011736 potassium bicarbonate Substances 0.000 claims description 4
- 229910000028 potassium bicarbonate Inorganic materials 0.000 claims description 4
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 claims description 4
- IDOQDZANRZQBTP-UHFFFAOYSA-N 2-[2-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol Chemical compound CC(C)(C)CC(C)(C)C1=CC=CC=C1OCCO IDOQDZANRZQBTP-UHFFFAOYSA-N 0.000 claims description 3
- 229920004929 Triton X-114 Polymers 0.000 claims description 3
- 239000008236 heating water Substances 0.000 claims description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims description 2
- 230000002055 immunohistochemical effect Effects 0.000 claims 1
- 239000003002 pH adjusting agent Substances 0.000 claims 1
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 abstract description 54
- 239000000126 substance Substances 0.000 abstract description 10
- 230000009286 beneficial effect Effects 0.000 abstract description 4
- 229910001385 heavy metal Inorganic materials 0.000 abstract description 4
- 231100000053 low toxicity Toxicity 0.000 abstract description 3
- 239000012188 paraffin wax Substances 0.000 description 9
- 239000003755 preservative agent Substances 0.000 description 7
- 230000002335 preservative effect Effects 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
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- 238000001514 detection method Methods 0.000 description 3
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- NOOLISFMXDJSKH-UTLUCORTSA-N (+)-Neomenthol Chemical compound CC(C)[C@@H]1CC[C@@H](C)C[C@@H]1O NOOLISFMXDJSKH-UTLUCORTSA-N 0.000 description 1
- GSFSVEDCYBDIGW-UHFFFAOYSA-N 2-(1,3-benzothiazol-2-yl)-6-chlorophenol Chemical compound OC1=C(Cl)C=CC=C1C1=NC2=CC=CC=C2S1 GSFSVEDCYBDIGW-UHFFFAOYSA-N 0.000 description 1
- 208000035404 Autolysis Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010057248 Cell death Diseases 0.000 description 1
- NOOLISFMXDJSKH-UHFFFAOYSA-N DL-menthol Natural products CC(C)C1CCC(C)CC1O NOOLISFMXDJSKH-UHFFFAOYSA-N 0.000 description 1
- 230000007035 DNA breakage Effects 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
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- 239000003795 chemical substances by application Substances 0.000 description 1
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- 238000004132 cross linking Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229960001484 edetic acid Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000003906 humectant Substances 0.000 description 1
- 238000013115 immunohistochemical detection Methods 0.000 description 1
- MGIYRDNGCNKGJU-UHFFFAOYSA-N isothiazolinone Chemical compound O=C1C=CSN1 MGIYRDNGCNKGJU-UHFFFAOYSA-N 0.000 description 1
- 229940041616 menthol Drugs 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000028043 self proteolysis Effects 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 231100000378 teratogenic Toxicity 0.000 description 1
- 230000003390 teratogenic effect Effects 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000004018 waxing Methods 0.000 description 1
Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/305—Fixative compositions
- G01N2001/307—Fixative compositions non-toxic, no Hg, no formaldehyde
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a novel formaldehyde-free tissue fixative and a preparation method thereof, and aims to provide a formaldehyde-free colorless and low-toxicity novel formaldehyde-free tissue fixative and a preparation method thereof, wherein the fixative comprises the following components in percentage by mass: 850-950 parts of tertiary butanol, 4-12 parts of fructose, 55-95 parts of weak acid, 10-60 parts of an epoxy ethylene condensate mixture, 2-10 parts of a detergent, 1-10 parts of a pH regulator, 1-5 parts of a metal ion chelating agent and 250 parts of water. The fixing solution does not contain harmful substances such as formaldehyde and the like, is beneficial to quick fixation of tissue samples, is added with a metal ion chelating agent to remove heavy metals possibly existing, and is more beneficial to observation results and ensures the reliability of the observation results.
Description
The technical field is as follows:
the invention belongs to the technical field of pathological experiments, and particularly relates to a colorless, low-toxicity, novel, formaldehyde-free and environment-friendly tissue fixative.
Background art:
in the field of biology, immobilization is intended to suppress destruction and autolysis of a living body or a part thereof, to preserve the shape, internal structure, substance composition, and the like of the living body as much as possible, or to dissolve a part of a substance and selectively preserve a part of the internal structure and the substance for the purpose of adaptation to studies, so as to observe the state of the living body. In addition, the operation steps such as embedding, slicing, staining and the like can be easily performed by fixing for the convenience of observation by an optical microscope and an electron microscope. Since the main components in the cell are protein and water, the fixation is mainly performed by the coagulation denaturation of protein by a fixative solution or heating or the freezing of water, and the fixation is performed by rapid cooling. Since fixation can artificially change cells, it is often necessary to take into account the change in cells when observing a material in a fixed state.
The tissue fixation in the pathological section process is important, the tissue is fixed by formalin (also called formaldehyde) fixing liquid in the prior art, but the formaldehyde fixing tissue has the defects of strong volatility, easy DNA breakage, easy protein crosslinking and the like, the formaldehyde is confirmed to be carcinogenic and teratogenic in China, and pathological operators are extremely bad for body health after being exposed to the formaldehyde environment for a long time.
Based on the above-mentioned shortcomings of formaldehyde fixation solution, finding a substitute for formaldehyde has become an urgent problem to be solved, and a part of formaldehyde substitutes are now available for tissue fixation. Patent publication No. CN102175496A discloses a formaldehyde-free tissue specimen fixing solution, and patent application publication No. CN104904706A discloses a formaldehyde-free animal specimen fixing solution and a preparation method thereof, but all contain acetone, and the acetone belongs to dangerous chemicals, is very corrosive, has pungent smell and is harmful to human bodies. Patent publication No. CN107367407A discloses a pathological specimen fixing and dehydrating treatment method, which comprises the following steps: fixing pathological specimen tissue, washing pathological specimen tissue, dehydrating pathological specimen tissue, infiltrating pathological specimen tissue, and waxing pathological specimen tissue, wherein the fixing solution comprises formaldehyde, water, sodium dihydrogen phosphate, disodium hydrogen phosphate, and formaldehyde. Patent application with publication number CN108760443A discloses a method for fixing pathological specimens, which adopts the raw materials of menthol, isothiazolinone and sodium dimethyldithiocarbamate, and the material is expensive and not commonly used, and is not easy to popularize in a large area. The patent application with the publication number of CN109805001A discloses a formaldehyde-free tissue fixation preservative solution and a preparation method thereof, the preservative solution comprises a protein fixing agent, a preservative, an osmotic pressure regulator, a humectant and an antioxidant, wherein the osmotic pressure regulator contains cane sugar which is easy to breed bacteria, mould and other microorganisms and is difficult to play a good preservative role; the other formaldehyde-free tissue fixing preservative solution emphasizes the preservative function rather than the fixing function, namely the invention is classified as the preservative solution.
The invention content is as follows:
in order to solve the defects of the fixing method, the invention discloses a novel formaldehyde-free tissue fixing solution, and aims to provide a formaldehyde-free colorless and low-toxicity novel formaldehyde-free tissue fixing solution and a preparation method thereof, wherein the novel formaldehyde-free tissue fixing solution is prepared by mixing tert-butyl alcohol, fructose, weak acid, an epoxy ethylene condensation compound mixture, a detergent, a pH regulator, a metal ion chelating agent and water, and comprises the following components in percentage by mass: 850-950 parts of tertiary butanol, 4-12 parts of fructose, 55-95 parts of weak acid, 10-60 parts of an epoxy ethylene condensate mixture, 2-10 parts of a detergent, 1-10 parts of a pH regulator, 1-5 parts of a metal ion chelating agent and 250 parts of water. The fixing solution does not contain harmful substances such as formaldehyde and the like, is beneficial to quick fixation of tissue samples, is added with a metal ion chelating agent to remove heavy metals possibly existing, and is more beneficial to observation results and ensures the reliability of the observation results.
The invention relates to a novel formaldehyde-free tissue fixative, which adopts the technical scheme that:
the novel formaldehyde-free tissue fixative is characterized by being a novel formaldehyde-free tissue fixative formed by mixing tert-butyl alcohol, fructose, weak acid, epoxy ethylene condensate mixture, a detergent, a pH regulator, a metal ion chelating agent and water, and comprising the following components in percentage by mass: 850-950 parts of tertiary butanol, 4-12 parts of fructose, 55-95 parts of weak acid, 10-60 parts of an epoxy ethylene condensate mixture, 2-10 parts of a detergent, 1-10 parts of a pH regulator, 1-5 parts of a metal ion chelating agent and 250 parts of water, wherein the fixing solution does not contain harmful substances such as formaldehyde and the like, is favorable for quickly fixing a tissue sample, and is more favorable for observing results and ensuring the reliability by adding the metal ion chelating agent to remove possible heavy metals.
The novel formaldehyde-free tissue fixative is characterized in that tertiary butanol is not lower than analytical purity grade, and fructose is not lower than biochemical reagent grade.
A novel formaldehyde-free tissue fixative is characterized in that the weak acid is one or a mixture of butyric acid and salicylic acid.
The preferable scheme is that salicylic acid is adopted as the weak acid.
The novel formaldehyde-free tissue fixative is characterized in that the epoxy ethylene condensate mixture is a mixture formed by mixing an epoxy ethylene condensate 1350 and an epoxy ethylene condensate 1500 according to the proportion of 1: 1-3, wherein the epoxy ethylene condensate 1350 and the epoxy ethylene condensate 1500 are milky solid at room temperature.
A novel formaldehyde-free tissue fixative is characterized in that the detergent is one or a mixture of two of Triton X-114 and polyoxyethylene sorbitan monostearate.
In the preferred embodiment, the detergent is Triton X-114 with a chemical purity level not lower than that.
The novel formaldehyde-free tissue fixative is characterized in that the pH regulator is one or a mixture of potassium carbonate and potassium bicarbonate.
The preferable scheme is that the potassium carbonate and the potassium bicarbonate both adopt the grade above AR.
The novel formaldehyde-free tissue fixative is characterized in that the metal ion chelating agent is one or a mixture of more of nitrilotriacetic acid, dihydroxyethylglycine and ethylene diamine tetraacetic acid.
Among them, it is preferable that the metal ion chelating agent is a mixture of nitrilotriacetic acid and dihydroxyethylglycine.
A novel formaldehyde-free tissue fixative is characterized in that water is not lower than experimental water level.
A novel formaldehyde-free tissue fixative is characterized in that the preparation method comprises the following operation steps: heating water to 75-95 deg.C, adding epoxy ethylene condensate mixture, dissolving, sequentially adding tert-butyl alcohol, fructose, weak acid, detergent, pH regulator and metal ion chelating agent, and mixing to obtain the novel formaldehyde-free tissue fixative.
A novel formaldehyde-free tissue fixative is characterized in that the fixative is used for fixing animal and plant samples.
A novel formaldehyde-free tissue fixative is characterized in that the fixative is used for routine pathological detection, immunohistochemical detection and molecular biological detection.
A novel formaldehyde-free tissue fixative is characterized in that the fixative can be used for clinical detection reagents and scientific research reagents.
The invention relates to a preparation method of novel formaldehyde-free tissue fixative, which comprises the following steps: heating water to 75-95 deg.C, adding epoxy ethylene condensate mixture, dissolving, sequentially adding tert-butyl alcohol, fructose, weak acid, detergent, pH regulator and metal ion chelating agent, mixing, and storing under sealed condition to obtain the novel formaldehyde-free tissue fixative.
When the novel formaldehyde-free tissue fixing solution is used for fixing, the fixing time and the fixing temperature can be properly adjusted by a person skilled in the art according to the fixing condition and requirements, and the fixing solution can be used for fixing samples such as animals, plants and the like.
For those skilled in the art, according to the technical scheme and concept of the present invention, the amount and ratio of the above components can be adjusted according to specific requirements, so as to prepare novel formaldehyde-free tissue fixative with different characteristics.
Compared with the prior art, the novel formaldehyde-free tissue fixative has the following advantages:
1. does not contain harmful substances such as formaldehyde and the like, and is not easy to cause harm to human bodies.
2. And adding a novel metal ion chelating agent to remove the possible heavy metal.
3. The fixing speed is high, and the fixation is complete.
Description of the drawings:
FIG. 1 is a schematic view of the treated tissue of the present invention "a novel formaldehyde-free tissue fixative" after paraffin embedding. Wherein, 1A is the result of visual observation after the paraffin embedding of the fixed tissue by using the novel formaldehyde-free tissue fixing solution, and 1B is the result of visual observation after the paraffin embedding of the fixed tissue by using the common formalin fixing solution.
FIG. 2 is a schematic view of a paraffin section HE staining microscope for treating tissues with the novel formaldehyde-free tissue fixative of the present invention. Wherein 2A is the observation result of the paraffin section HE staining microscope for fixing tissues by using the novel formaldehyde-free tissue fixing solution in the second embodiment, and 2B is the observation result of the paraffin section HE staining microscope for fixing tissues by using the common formalin fixing solution in the second embodiment.
The specific implementation mode is as follows:
the following detailed description further illustrates the invention as "a novel formaldehyde-free tissue fixative solution," but should not be construed as limiting the invention. Modifications or substitutions to methods, conditions, steps and applications of the invention may be made without departing from the spirit and substance of the invention.
Example one
The invention relates to a novel formaldehyde-free tissue fixing solution which is used for fixing animal tissues and comprises the following components in percentage by mass: 870 parts of tertiary butanol, 5 parts of fructose, 70 parts of salicylic acid, 20 parts of an epoxy ethylene condensate mixture, 2 parts of a mixture of potassium carbonate and potassium bicarbonate, 2 parts of nitrilotriacetic acid and 180 parts of water, and the mixture is mixed and fully mixed. The following examples use conventional formalin fixatives at the same treatment time, tissue, and temperature as controls.
The preparation method of the novel formaldehyde-free tissue fixative comprises the following steps: heating 180 parts of water to 75-95 ℃, adding 20 parts of epoxy ethylene condensation compound mixture, fully dissolving, sequentially mixing 870 parts of tert-butyl alcohol, 5 parts of fructose, 70 parts of salicylic acid, Triton X-1143 parts, 4 parts of potassium carbonate and 2 parts of nitrilotriacetic acid, fully mixing, and hermetically storing to obtain the novel formaldehyde-free tissue fixative.
Taking the novel formaldehyde-free tissue fixing solution prepared by the steps, and fixing the same tissue for 3 hours; taking a conventional formalin fixing solution, and fixing the same animal tissue for 3 hours; the results are shown in FIG. 1.
The experimental results show that, under the same conditions, the novel formaldehyde-free tissue fixative of the first embodiment is compared with the conventional formalin fixative, the fixation effect has no obvious difference, and the results are basically consistent.
Example two
The invention relates to a novel formaldehyde-free tissue fixing solution which is used for fixing animal tissues and comprises the following components in percentage by mass: 910 parts of tert-butyl alcohol, 7 parts of fructose, 65 parts of butyric acid, 12 parts of an epoxy ethylene condensation compound mixture, 2 parts of polyoxyethylene sorbitan monostearate, 3 parts of potassium carbonate, 3 parts of dihydroxyethyl glycine and 210 parts of water, and the components are mixed in sequence and fully and uniformly mixed. The following examples use conventional formalin fixatives at the same treatment time, tissue, and temperature as controls.
The preparation method of the novel formaldehyde-free tissue fixative comprises the following steps: heating 210 parts of water to 75-95 ℃, adding 12 parts of an epoxy ethylene condensation compound mixture, fully dissolving, sequentially mixing 910 parts of tert-butyl alcohol, 7 parts of fructose, 65 parts of butyric acid, 2 parts of polyoxyethylene sorbitan monostearate, 3 parts of potassium carbonate and 3 parts of dihydroxyethyl glycine, fully mixing, and hermetically storing to obtain the novel formaldehyde-free tissue fixative.
Taking the novel formaldehyde-free tissue fixing solution prepared by the steps, fixing the same tissue for 4 hours, and staining a paraffin section HE of the tissue; taking a conventional formalin fixing solution, fixing the same animal tissue for 4 hours, and staining a paraffin section HE of the tissue; the results are shown in FIG. 2.
The experimental result shows that when the novel formaldehyde-free tissue fixing solution of the second embodiment is compared with the conventional formalin fixing solution under the same condition when the tissue paraffin section is subjected to HE staining, the HE staining effect has no obvious difference and is basically consistent.
It is obvious to those skilled in the art that the time and temperature for fixing the stationary liquid can be adjusted according to the specific needs according to the technical scheme and concept of the present invention, and obvious changes or modifications therefrom should fall within the protection scope of the present invention.
Claims (10)
1. The novel formaldehyde-free tissue fixative is characterized by being a novel formaldehyde-free tissue fixative formed by mixing tert-butyl alcohol, fructose, weak acid, epoxy ethylene condensate mixture, a detergent, a pH regulator, a metal ion chelating agent and water, and comprising the following components in percentage by mass: 850-950 parts of tertiary butanol, 4-12 parts of fructose, 55-95 parts of weak acid, 10-60 parts of an epoxy ethylene condensate mixture, 2-10 parts of a detergent, 1-10 parts of a pH regulator, 1-5 parts of a metal ion chelating agent and 250 parts of water.
2. The novel formaldehyde-free tissue fixative as claimed in claim 1, wherein t-butanol is not less than analytical grade t-butanol and fructose is not less than biochemical grade fructose.
3. The novel formaldehyde-free tissue fixative as claimed in claim 1, wherein the weak acid is butyric acid, salicylic acid or a mixture thereof.
4. The novel formaldehyde-free tissue fixative solution of claim 1, wherein the mixture of epoxy ethylene condensate is a mixture of epoxy ethylene condensate 1350 and epoxy ethylene condensate 1500 mixed in a ratio of 1: 1-3, wherein the epoxy ethylene condensate 1350 and the epoxy ethylene condensate 1500 should be milky white solids at room temperature.
5. The novel formaldehyde-free tissue fixative according to claim 1, wherein the detergent is one or a mixture of Triton X-114 and polyoxyethylene sorbitan monostearate.
6. The novel formaldehyde-free tissue fixative as claimed in claim 1, wherein the pH adjusting agent is one or a mixture of potassium carbonate and potassium bicarbonate, and the metal ion chelating agent is one or a mixture of nitrilotriacetic acid, dihydroxyethylglycine and ethylenediaminetetraacetic acid.
7. The novel formaldehyde-free tissue fixative as claimed in claim 1, wherein the water is not lower than experimental water grade.
8. The novel formaldehyde-free tissue fixative according to claim 1, wherein the preparation method comprises the following steps: heating water to 75-95 deg.C, adding epoxy ethylene condensation compound mixture, dissolving, sequentially adding tert-butyl alcohol, fructose, weak acid, detergent and pH regulator, and mixing to obtain novel formaldehyde-free tissue fixative.
9. The novel formaldehyde-free tissue fixative as claimed in claim 1, wherein the fixative is used for fixation of animal and plant samples.
10. The novel formaldehyde-free tissue fixative as claimed in claim 1, wherein the fixative is used for routine pathological examination, immunohistochemical examination, molecular biology examination, clinical examination reagents and scientific reagents.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911117669.8A CN110779785A (en) | 2019-11-15 | 2019-11-15 | Novel formaldehyde-free tissue fixing solution |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201911117669.8A CN110779785A (en) | 2019-11-15 | 2019-11-15 | Novel formaldehyde-free tissue fixing solution |
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| CN110779785A true CN110779785A (en) | 2020-02-11 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111397999A (en) * | 2020-05-09 | 2020-07-10 | 中国科学院植物研究所 | Simple method for enhancing contrast of micro-CT plant sample |
| CN111397997A (en) * | 2020-04-26 | 2020-07-10 | 中烨(山东)检验检测有限公司 | Tissue fixing liquid for fixing fresh tissue sample |
| CN113519505A (en) * | 2021-09-01 | 2021-10-22 | 宁波西敦医药包衣科技有限公司 | Environment-friendly water-based cross-linking agent biological tissue preservation material and preparation method thereof |
-
2019
- 2019-11-15 CN CN201911117669.8A patent/CN110779785A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111397997A (en) * | 2020-04-26 | 2020-07-10 | 中烨(山东)检验检测有限公司 | Tissue fixing liquid for fixing fresh tissue sample |
| CN111397999A (en) * | 2020-05-09 | 2020-07-10 | 中国科学院植物研究所 | Simple method for enhancing contrast of micro-CT plant sample |
| CN113519505A (en) * | 2021-09-01 | 2021-10-22 | 宁波西敦医药包衣科技有限公司 | Environment-friendly water-based cross-linking agent biological tissue preservation material and preparation method thereof |
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