CN110455602A - Specific stain liquid and its preparation method and application based on cast-off cells dyeing - Google Patents
Specific stain liquid and its preparation method and application based on cast-off cells dyeing Download PDFInfo
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- CN110455602A CN110455602A CN201910649666.2A CN201910649666A CN110455602A CN 110455602 A CN110455602 A CN 110455602A CN 201910649666 A CN201910649666 A CN 201910649666A CN 110455602 A CN110455602 A CN 110455602A
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- dyeing
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- 238000004043 dyeing Methods 0.000 title claims abstract description 70
- 239000007788 liquid Substances 0.000 title claims abstract description 66
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 37
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims abstract description 35
- 230000000149 penetrating effect Effects 0.000 claims abstract description 29
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 claims abstract description 28
- 229960000907 methylthioninium chloride Drugs 0.000 claims abstract description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 235000019152 folic acid Nutrition 0.000 claims abstract description 19
- 239000011724 folic acid Substances 0.000 claims abstract description 19
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims abstract description 16
- 229960000304 folic acid Drugs 0.000 claims abstract description 16
- 239000003638 chemical reducing agent Substances 0.000 claims abstract description 14
- 239000003002 pH adjusting agent Substances 0.000 claims abstract description 12
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 45
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 20
- 239000000243 solution Substances 0.000 claims description 20
- 235000017166 Bambusa arundinacea Nutrition 0.000 claims description 15
- 235000017491 Bambusa tulda Nutrition 0.000 claims description 15
- 241001330002 Bambuseae Species 0.000 claims description 15
- 235000015334 Phyllostachys viridis Nutrition 0.000 claims description 15
- 239000011425 bamboo Substances 0.000 claims description 15
- 239000000052 vinegar Substances 0.000 claims description 15
- 235000021419 vinegar Nutrition 0.000 claims description 15
- 229920000742 Cotton Polymers 0.000 claims description 11
- 235000011187 glycerol Nutrition 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 10
- CDQSJQSWAWPGKG-UHFFFAOYSA-N butane-1,1-diol Chemical compound CCCC(O)O CDQSJQSWAWPGKG-UHFFFAOYSA-N 0.000 claims description 9
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 6
- 125000003929 folic acid group Chemical group 0.000 claims description 5
- 210000000981 epithelium Anatomy 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 abstract description 45
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 abstract description 44
- 235000010323 ascorbic acid Nutrition 0.000 abstract description 21
- 239000011668 ascorbic acid Substances 0.000 abstract description 21
- 229960005070 ascorbic acid Drugs 0.000 abstract description 21
- 210000000170 cell membrane Anatomy 0.000 abstract description 4
- 230000007062 hydrolysis Effects 0.000 abstract description 3
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 3
- 230000008591 skin barrier function Effects 0.000 abstract description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 21
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 10
- 238000000034 method Methods 0.000 description 9
- 206010028980 Neoplasm Diseases 0.000 description 8
- 201000011510 cancer Diseases 0.000 description 7
- 210000001519 tissue Anatomy 0.000 description 7
- 238000004040 coloring Methods 0.000 description 6
- 239000004310 lactic acid Substances 0.000 description 5
- 235000014655 lactic acid Nutrition 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 239000002904 solvent Substances 0.000 description 4
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 229940064302 folacin Drugs 0.000 description 3
- 208000014674 injury Diseases 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- GDSOZVZXVXTJMI-SNAWJCMRSA-N (e)-1-methylbut-1-ene-1,2,4-tricarboxylic acid Chemical compound OC(=O)C(/C)=C(C(O)=O)\CCC(O)=O GDSOZVZXVXTJMI-SNAWJCMRSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- FBWADIKARMIWNM-UHFFFAOYSA-N N-3,5-dichloro-4-hydroxyphenyl-1,4-benzoquinone imine Chemical compound C1=C(Cl)C(O)=C(Cl)C=C1N=C1C=CC(=O)C=C1 FBWADIKARMIWNM-UHFFFAOYSA-N 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 235000010338 boric acid Nutrition 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 230000005101 cell tropism Effects 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 239000006184 cosolvent Substances 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002083 enediols Chemical group 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- QTWZICCBKBYHDM-UHFFFAOYSA-N leucomethylene blue Chemical compound C1=C(N(C)C)C=C2SC3=CC(N(C)C)=CC=C3NC2=C1 QTWZICCBKBYHDM-UHFFFAOYSA-N 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- KSBAEPSJVUENNK-UHFFFAOYSA-L tin(ii) 2-ethylhexanoate Chemical compound [Sn+2].CCCCC(CC)C([O-])=O.CCCCC(CC)C([O-])=O KSBAEPSJVUENNK-UHFFFAOYSA-L 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
Classifications
-
- 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
- G01N2001/302—Stain compositions
Landscapes
- 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)
Abstract
The invention discloses a kind of specific stain liquid and its preparation method and application based on cast-off cells dyeing.By weight percentage, which includes: folic acid 2-5%, reducing agent 1-12%, pH adjusting agent 2-8%, methylene blue 2-5%, penetrating agent 3-25%, water 5-85%.The present invention is added to penetrating agent in specific stain liquid, and penetrating agent can not only slow down the hydrolysis of the reducing agents such as ascorbic acid, it is ensured that methylene blue is in reduction-state, improves the stability of specific stain liquid；Methylene blue can also be promoted to be dyed quickly through skin barrier and cell membrane, improve dyeing efficiency and dyeing accuracy rate, avoid the appearance of false positive results.
Description
Technical field
The invention belongs to living cells coloring agent technical fields, and in particular to a kind of specific stain based on cast-off cells dyeing
Liquid and its preparation method and application.
Background technique
Specific stain is a kind of to show and occur in the normal configuration or pathologic process that determine in tissue or cell
Exotic matter, lesion and pathogen etc., and corresponding dyeing liquor is selected to show the colouring method of these ingredients.
Clinical research shows that the quantity of folacin receptor can be expressed in cancer cell surfaces height, and in normal cell then compared with
Less or do not express；Simultaneously because cancer cell is in strong oxidative stress environment, therefore there is also a large amount of living in cancer cell
Property oxygen, and then without this phenomenon in normal cell.There is a kind of new Staining for Diagnosis side on the basis of the clinical research
The compound that method, i.e., folacin receptor mediated specific stain method, methylene blue and folic acid are formed has efficient target to cancer cell
Tropism, and the folacin receptor that the compound is easier to cancer cell surfaces in acid condition combines, so as to enter cancer cell；And
Under the oxidative stress environment of tumour cell, the leucomethylene blue of reduction-state can be converted and blue or blue-black oxidation state,
It is achieved in the diagnosis to cancer cell.
Based on the principle of the special color method, occur many relevant specific stain liquid on the market at present.Such as disclosure
It number is combined to disclose a kind of coloring agent for tumor tissue cell in the Chinese invention patent application of 103483872 A of CN
Object, by weight percentage, the coloring composition include: color developing agent 0.01-5%, folic acid≤6.5%, procrypsis agent 0.1-
10%, pH adjusting agent≤10%, water 3-95%；Wherein, color developing agent is in methylene blue, dichloropheno-lindophenol and dichlorophenol indophenol sodium
At least one, procrypsis agent be at least one of ascorbic acid, zinc powder, stannous octoate, pH adjusting agent be formic acid, acetic acid, cream
Acid, citric acid, malic acid, oxalic acid, succinic acid, tartaric acid or boric acid.
In the prior art frequently with ascorbic acid as procrypsis agent, however the stability of ascorbic acid is poor, this will lead
The diagnostic result of dyeing liquor is caused error occur；In order to be avoided as much as the error, current this kind of specific stain liquid is being used
When, all it is that dyeing liquor is first first dipped using carrier (such as cotton swab), dyeing liquor is applied directly to tissue, then observes carrier
The color change of upper dyeing liquor judges according to the color change whether the histocyte region tested generates lesion.
This application method is disadvantageous in that 1. the time of dyeing liquor ingress of air is longer, and the methylene blue of reproducibility is easy to
Oxidation by air and there is false positive diagnostic result；2. dyeing liquor directly contacts human body, it is easy to generate toxicity to human body, and causes
Secondary injury.
Summary of the invention
Present invention purpose be to provide a kind of specific stain liquid and preparation method thereof based on cast-off cells dyeing and
Using.
For achieving the above object, the technical solution of the application is as follows:
A kind of specific stain liquid based on cast-off cells dyeing, by weight percentage, comprising: folic acid 2-5%, reducing agent
1-12%, pH adjusting agent 2-8%, methylene blue 2-5%, penetrating agent 3-25%, water 5-85%.
The present invention is added to penetrating agent in specific stain liquid, and penetrating agent can not only slow down the reducing agents such as ascorbic acid
Hydrolysis, it is ensured that methylene blue is in reduction-state, improves the stability of specific stain liquid；Methylene blue can also be promoted quickly through skin
Skin barrier and cell membrane are dyed, and are improved dyeing efficiency and dyeing accuracy rate, are avoided the appearance of false positive results.
In the present invention, the reducing agent can be ascorbic acid or zinc powder, the pH adjusting agent can be acetic acid or
Lactic acid.
It is above-mentioned based on cast-off cells dyeing specific stain liquid in, the penetrating agent be propylene glycol, glycerine and
At least one of butanediol.Ring composition in ascorbic acid structure is enediol base, and one is σ key in carbon-carbon double bond, another
A is pi bond；Wherein, pi bond easily causes conjugation, and α carbon electronics forms π (2,2) in p track, at the same with two oxygen being connected
The pi-conjugated effect π (4,6) of p- occurs for atom, and electronegativity enhancing is easy to decline H atom attraction, hydrolyze；Meanwhile anti-
In the aqueous solution of bad hematic acid, there are secondary key-hydrogen bond between hydrone, and hydrogen atom and ortho position carbonyl in ascorbic acid molecule structure
Base oxygen atom can then generate the secondary key slightly different with water, be easy to make lactone cyclizing hydrolysis.Propylene glycol, glycerine and butanediol etc.
The lesser solvent of polarity can destroy the secondary key, reach and slow down the acid-hydrolyzed purpose of Vitamin C.
In the above-mentioned specific stain liquid based on cast-off cells dyeing, by weight percentage, comprising: folic acid 3-4%,
Reducing agent 6-9%, pH adjusting agent 2-8%, methylene blue 3-4%, penetrating agent 14-20%, water 5-75%.
It is described in the specific stain liquid of the invention based on cast-off cells dyeing in addition to using above-mentioned penetrating agent
Penetrating agent can also include:
1.: at least one of propylene glycol, glycerine and butanediol；
2.: bamboo vinegar.
Bamboo vinegar not only has anti-microbial property, but also because it contains more organic acid substance, letones and alcohols
Substance, therefore can make to be dissolved in bamboo vinegar as good bleeding agent as good solvent, solubilizer or cosolvent
Folic acid and methylene blue can effectively permeate, spread, pass through cell membrane realize rapid dyeing.Bamboo vinegar and propylene glycol etc. have
When solvent is combined, the dyeing efficiency and accuracy rate of specific stain liquid be can be further improved.
In the above-mentioned specific stain liquid based on cast-off cells dyeing, by weight percentage, the penetrating agent packet
It includes:
1. at least one of propylene glycol, glycerine and butanediol: 80-95%；
2. bamboo vinegar: 5-20%.
In the above-mentioned specific stain liquid based on cast-off cells dyeing by weight percentage, the penetrating agent includes
Propylene glycol 87% and bamboo vinegar 13%.
In the above-mentioned specific stain liquid based on cast-off cells dyeing, by weight percentage, comprising: folic acid 3-4%, also
Former agent 3-5%, pH adjusting agent 2-5%, methylene blue 3-4%, penetrating agent 7-12%, water 5-80%.When containing bamboo in penetrating agent
When vinegar liquid, the dosage of penetrating agent and reducing agent can be reduced suitably.
It is of the present invention based on cast-off cells dyeing specific stain liquid preparation method the following steps are included:
(1) preset weight percent is pressed, will be added in penetrating agent, stir evenly after folic acid part aqueous solution；
(2) preset weight percent is pressed, the reducing agent of 50-70% is added in the solution of step (1) acquisition, stirring
Uniformly；
(3) preset weight percent is pressed, methylene blue is added in the solution of step (2) acquisition, 15- is stirred
45min；
(4) preset weight percent is pressed, pH adjusting agent and remaining reducing agent are added to the solution of step (3) acquisition
In, after mixing evenly, with water constant volume, obtain the specific stain liquid based on cast-off cells dyeing.
Specific stain liquid based on cast-off cells dyeing answering in the detection of epithelial tissue cancerous tumor cell of the present invention
With including:
(a) it is enclosed with cotton swab in suspected location surface wipes 3-5, to pick cast-off cells；
(b) the specific stain liquid based on cast-off cells dyeing is dipped with the cotton swab for picking cast-off cells, observes cotton
The color change of label.
When using specific stain liquid dyeing of the invention, first tissue can be taken to dye again, having broken in conventional method needs
Dyeing liquor is applied directly to the natural mode of tissue, the time that dyeing liquor is contacted with air is shorter, and testing result is quasi-
True rate is higher；And to patient close friend, patient compliance more preferably, avoids secondary injury.
Compared with prior art, the beneficial effects of the present invention are embodied in:
(1) present invention is added to penetrating agent in specific stain liquid, and penetrating agent can not only slow down the reduction such as ascorbic acid
The hydrolysis of agent, it is ensured that methylene blue is in reduction-state, improves the stability of specific stain liquid；Methylene blue can also be promoted quickly to lead to
It crosses skin barrier and cell membrane is dyed, improve dyeing efficiency and dyeing accuracy rate, avoid the appearance of false positive results.
(2) when using specific stain liquid dyeing of the invention, first tissue can be taken to dye again, broken in conventional method
Need for dyeing liquor to be applied directly to the natural mode of tissue, the time that dyeing liquor is contacted with air is shorter, testing result
Accuracy rate is higher；And to patient close friend, patient compliance more preferably, avoids secondary injury.
Specific embodiment
Specific embodiment is set forth below, and further details of the technical solution of the present invention.
Embodiment 1
A kind of specific stain liquid based on cast-off cells dyeing of the present embodiment, by weight percentage, comprising: folic acid 3%,
Ascorbic acid 6%, acetic acid 4%, methylene blue 3%, propylene glycol 14%, water 70%.
This based on cast-off cells dyeing specific stain liquid preparation method the following steps are included:
(1) preset weight percent is pressed, will be added in propylene glycol after folic acid part aqueous solution, is stirred to completely molten
Solution；
(2) preset weight percent is pressed, 50% ascorbic acid is added in the solution of step (1) acquisition, stirring
Uniformly；
(3) preset weight percent is pressed, methylene blue is added in the solution of step (2) acquisition, stirring 30min is extremely
It is completely dissolved；
(4) preset weight percent is pressed, acetic acid and remaining ascorbic acid are added to the solution of step (3) acquisition
In, after mixing evenly, with water constant volume, obtain the specific stain liquid based on cast-off cells dyeing of the present embodiment.
Embodiment 2
A kind of specific stain liquid based on cast-off cells dyeing of the present embodiment, by weight percentage, comprising: folic acid 4%,
Ascorbic acid 9%, lactic acid 6%, methylene blue 4%, glycerine 20%, water 57%.
This based on cast-off cells dyeing specific stain liquid preparation method the following steps are included:
(1) preset weight percent is pressed, will be added in propylene glycol after folic acid part aqueous solution, is stirred to completely molten
Solution；
(2) preset weight percent is pressed, 60% ascorbic acid is added in the solution of step (1) acquisition, stirring
Uniformly；
(3) preset weight percent is pressed, methylene blue is added in the solution of step (2) acquisition, stirring 45min is extremely
It is completely dissolved；
(4) preset weight percent is pressed, lactic acid and remaining ascorbic acid are added to the solution of step (3) acquisition
In, after mixing evenly, with water constant volume, obtain the specific stain liquid based on cast-off cells dyeing of the present embodiment.
Embodiment 3
A kind of specific stain liquid based on cast-off cells dyeing of the present embodiment, by weight percentage, comprising: folic acid 4%,
Ascorbic acid 5%, acetic acid 2%, methylene blue 4%, penetrating agent 12%, water 73%；
By weight percentage, which is made of propylene glycol 87% and bamboo vinegar 13%.
This based on cast-off cells dyeing specific stain liquid preparation method the following steps are included:
(1) preset weight percent is pressed, will be added in propylene glycol after folic acid part aqueous solution, is stirred to completely molten
Solution；
(2) preset weight percent is pressed, 50% ascorbic acid is added in the solution of step (1) acquisition, stirring
Uniformly；
(3) preset weight percent is pressed, methylene blue is added in the solution of step (2) acquisition, stirring 20min is extremely
It is completely dissolved；
(4) preset weight percent is pressed, acetic acid and remaining ascorbic acid are added to the solution of step (3) acquisition
In, after mixing evenly, with water constant volume, obtain the specific stain liquid based on cast-off cells dyeing of the present embodiment.
Embodiment 4
A kind of specific stain liquid based on cast-off cells dyeing of the present embodiment, by weight percentage, comprising: folic acid 3%,
Ascorbic acid 3%, lactic acid 5%, methylene blue 3%, penetrating agent 7%, water 79%；
By weight percentage, which is made of glycerine 95% and bamboo vinegar 5%.
The preparation method of the specific stain liquid based on cast-off cells dyeing is same as Example 3.
Embodiment 5
A kind of specific stain liquid based on cast-off cells dyeing of the present embodiment, by weight percentage, comprising: folic acid 3%,
Ascorbic acid 4%, lactic acid 4%, methylene blue 4%, penetrating agent 10%, water 75%；
By weight percentage, which is made of butanediol 90% and bamboo vinegar 10%.
The preparation method of the specific stain liquid based on cast-off cells dyeing is same as Example 3.
Comparative example 1
A kind of specific stain liquid based on cast-off cells dyeing of the present embodiment, by weight percentage, comprising: folic acid 4%,
Ascorbic acid 5%, acetic acid 2%, methylene blue 4%, bamboo vinegar 12%, water 73%.
The preparation method of the specific stain liquid based on cast-off cells dyeing is same as Example 3.
Specific stain liquid made from Example 1-5, comparative example 1, while it is special to have purchased existing Epithelial cell
Dyeing liquor is tested the stability and usage mode of each specific stain liquid, and test result is shown in Table 1.
Wherein, the test method of stability are as follows: every kind of specific stain liquid takes 4 parts, dips dyeing liquor with cotton swab, rests on
In air, stopwatch records the Coloring Time of dyeing liquor, is averaged.
Table 1
Seen from table 1, the average Coloring Time of the specific stain liquid of embodiment 1-2 preparation is prepared in 55-56s, comparative example 1
Specific stain liquid average Coloring Time in 50s, and the average Coloring Time of the specific stain liquid of embodiment 3-5 preparation exists
64-68s, it is stronger with each specific stain liquid phase ratio, the stability of specific stain liquid of the invention in the prior art；Meanwhile third
The compatible use of glycol, glycerine and butanediol and bamboo vinegar can further increase the stability of dye liquor.
Also, the application method of specific stain liquid of the invention are as follows: first enclosed with cotton swab in suspected location surface wipes 3-5,
To pick cast-off cells；Specific stain liquid then is dipped with the cotton swab for picking cast-off cells, observes the color change of cotton swab.This with
Each specific stain liquid needs first to dip in that dyeing liquor, that dyeing liquor is smeared to the application method sampled again with suspected location is complete in the prior art
Complete different, in application method of the invention, the time that dyeing liquor is contacted with air is shorter, and the accuracy rate of testing result is higher, and
Dyeing liquor will not directly contact human body.
The detection sensitivity and specificity of specific stain liquid listed in table 1 are detected, detection method are as follows:
Sensitivity=true positives number/(true positives number+false positive number) × 100%；
Specificity=true negative number/(true negative number+false positive number) × 100%；
Testing result is shown in Table 2.
Table 2
As can be seen from Table 2, specific stain liquid prepared by the present invention all has higher detection sensitivity and specificity, show have
There is higher Detection accuracy.
Claims (10)
1. a kind of specific stain liquid based on cast-off cells dyeing, which is characterized in that by weight percentage, comprising: folic acid 2-
5%, reducing agent 1-12%, pH adjusting agent 2-8%, methylene blue 2-5%, penetrating agent 3-25%, water 5-85%.
2. the specific stain liquid as described in claim 1 based on cast-off cells dyeing, which is characterized in that the penetrating agent is
At least one of propylene glycol, glycerine and butanediol.
3. the specific stain liquid as claimed in claim 2 based on cast-off cells dyeing, which is characterized in that by weight percentage
Meter, comprising: folic acid 3-4%, reducing agent 6-9%, pH adjusting agent 2-8%, methylene blue 3-4%, penetrating agent 14-20%, water 5-
75%.
4. the specific stain liquid as described in claim 1 based on cast-off cells dyeing, which is characterized in that the penetrating agent packet
It includes:
1.: at least one of propylene glycol, glycerine and butanediol；
2.: bamboo vinegar.
5. the specific stain liquid as claimed in claim 4 based on cast-off cells dyeing, which is characterized in that by weight percentage
Meter, the penetrating agent include:
1. at least one of propylene glycol, glycerine and butanediol: 80-95%；
2. bamboo vinegar: 5-20%.
6. the specific stain liquid as claimed in claim 5 based on cast-off cells dyeing, which is characterized in that by weight percentage
Meter, the penetrating agent includes propylene glycol 87% and bamboo vinegar 13%.
7. the specific stain liquid based on cast-off cells dyeing as described in any one of claim 4-6, which is characterized in that press
Weight percent meter, comprising: folic acid 3-4%, reducing agent 3-5%, pH adjusting agent 2-5%, methylene blue 3-4%, penetrating agent 7-
12%, water 5-80%.
8. the preparation method of the specific stain liquid based on cast-off cells dyeing as described in any one of claim 1-7,
It is characterized in that, comprising the following steps:
(1) preset weight percent is pressed, will be added in penetrating agent, stir evenly after folic acid part aqueous solution；
(2) preset weight percent is pressed, the reducing agent of 50-70% is added in the solution of step (1) acquisition, stirring is equal
It is even；
(3) preset weight percent is pressed, methylene blue is added in the solution of step (2) acquisition, 15-45min is stirred；
(4) preset weight percent is pressed, pH adjusting agent and remaining reducing agent are added in the solution of step (3) acquisition,
After mixing evenly, with water constant volume, the specific stain liquid based on cast-off cells dyeing is obtained.
9. the specific stain liquid based on cast-off cells dyeing as described in any one of claim 1-7 is in epithelial tissue canceration
Application in cell detection.
10. application as claimed in claim 9 characterized by comprising
(a) it is enclosed with cotton swab in suspected location surface wipes 3-5, to pick cast-off cells；
(b) dyeing as described in any one of claim 1-7 based on cast-off cells is dipped with the cotton swab for picking cast-off cells
Specific stain liquid, observe the color change of cotton swab.
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Application publication date: 20191115 |