CN105067412B - A kind of vaginal fluid dyeing liquor and preparation method thereof and colouring method - Google Patents
A kind of vaginal fluid dyeing liquor and preparation method thereof and colouring method Download PDFInfo
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- CN105067412B CN105067412B CN201510484688.XA CN201510484688A CN105067412B CN 105067412 B CN105067412 B CN 105067412B CN 201510484688 A CN201510484688 A CN 201510484688A CN 105067412 B CN105067412 B CN 105067412B
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Abstract
The present invention discloses a kind of vaginal fluid dyeing liquor and preparation method thereof and colouring method, the present invention using blue color basic dye and rosamines due to being mixed to prepare dyeing liquor, so that dyeing liquor obtained has the coloring of general dyeing liquor, and dyeing time can be greatly shortened, the dyeing time usually completed in 5 ~ 30 minutes is made to shorten to 20s, staining efficiency is greatly improved, accelerates the screening speed of clinical sample.In addition, using the image clearly after dyeing liquor of the present invention dyeing, it is easily recognized, can clearly identifies surface layer squamous cell, middle level epithelial cell, bottom epithelial cell, clues cell, leucocyte, the ingredients such as red blood cell, Candida albicans, trichomonad, suitable for the Clinical screening of great amount of samples.
Description
Technical field
The present invention relates to cell dyeing technical field more particularly to a kind of vaginal fluid dyeing liquor and preparation method thereof with
Colouring method.
Background technology
Endonuclear chromatin is mainly DNA（DNA）, in DNA double helical structure, on two chains
Phosphate is negatively charged towards outside, easily with positively charged basic dye with ionic bond or Hydrogenbond, so as to make dyeing
Matter adheres to corresponding color.Nucleus is chromatinic storage place, and chromatin is fine and close, then nucleus color depth；Chromatin is dredged
Pine, then nucleus coloring is shallow.Albumen iso-electric point is 3~6 in secretion cytoplasm, works as pH>During PI, free carboxyl in albumen
Group increases, and can be combined with positively charged basic dye, so as to which cytoplasm be made to adhere to corresponding color.Cytoplasm institute band is negative
Charge is more, then cytoplasm dyeing is relatively deep；Cytoplasm institute is negatively charged few, then cytoplasm dyeing is relatively shallower.And due to
Cell and asynchronous proliferation in secretion, therefore the cell color degree within the different cell cycles has differences.It is in
For the cell of growth period and division stage since DNA and protein synthesis are vigorous and nucleus is presented and color depth, cytoplasm coloring is opposite
The phenomenon that shallower.And the cell in apoptosis period is since permeability of cell membrane is strengthened, acidic amino acid site is sudden and violent in protein
Dew, chromatin dna segment degradation and make cell feulgen's stain distinguish unobvious.
At present, the relatively broad vaginal fluid dyeing liquor of domestic application is mainly that Rui Shi-Ji's nurse Sa dyeing liquor and leather are blue
Albert'stain Albert liquid, but both dyeing liquors all have the shortcomings that complex for operation step, dyeing time is long, and being unfavorable for larger scale clinical should
With.
Therefore, the prior art has yet to be improved and developed.
Invention content
In view of above-mentioned deficiencies of the prior art, the purpose of the present invention is to provide a kind of vaginal fluid dyeing liquor and its systems
Preparation Method and colouring method, it is intended to which solving existing dyeing liquor, there are complex for operation step, dyeing time are long and be unfavorable for extensive
The problem of clinical practice.
Technical scheme is as follows：
A kind of preparation method of vaginal fluid dyeing liquor, wherein, including step：
A, by weight, 1 ~ 3 part of blue color basic dye, 6 ~ 8 parts of rosamines, 1 ~ 3 part of buffer salt are weighed respectively,
8 ~ 12 parts of alcoholic solvent, 75 ~ 85 parts of deionized water；
B, by the above-mentioned blue color basic dye weighed, rosamines, buffer salt, alcoholic solvent and deionized water 22 ~
It is uniformly mixed, and stands overnight at 28 DEG C；
C, 0.02 ~ 0.5 part of alkaline matter, membrane protective agent 0.01 ~ 0.5 are added in the mixed liquor after standing overnight
Part, 0.01 ~ 0.5 part of dyeing promoter, 0.002 ~ 0.2 part of surfactant, 0.01 ~ 0.5 part of osmotic pressure regulator, it is finally quiet again
It puts overnight, obtains dyeing liquor.
The preparation method of the vaginal fluid dyeing liquor, wherein, the blue color basic dye is methylenum careuleum, reddish black
It is one or more in II, methyl green, crystal violet, toluidine blue.
The preparation method of the vaginal fluid dyeing liquor, wherein, the rosamines for dimethyl diaminophenazine chloride, safranine T,
It is one or more in eosin Y, methyl red.
The preparation method of the vaginal fluid dyeing liquor, wherein, the alkaline matter is natrium carbonicum calcinatum, bicarbonate
It is one or more in sodium, potassium acetate, sodium acetate, calcium acetate, borax.
The preparation method of the vaginal fluid dyeing liquor, wherein, the membrane protective agent for trehalose, sucrose,
It is one or more in mannitol, glycine.
The preparation method of the vaginal fluid dyeing liquor, wherein, the dyeing promoter is cytase, mucus
It is one or more in distintegrant bromhexine, ambroxol, acetylcysteine.
The preparation method of the vaginal fluid dyeing liquor, wherein, the surfactant is alkylphenol-polyethenoxy
Ether, high-carbon fatty alcohol polyoxyethylene ether, polyoxyethylene carboxylate, fatty acid methyl ester ethoxylate, polypropylene glycol epoxy
It is one or more in ethane additive product, sorbitan ester, sucrose ester.
The preparation method of the vaginal fluid dyeing liquor, wherein, the osmotic pressure regulator is glycerine, PVP, grape
It is one or more in sugar, fructose, polyglucose.
A kind of vaginal fluid dyeing liquor, which is characterized in that using as above any vaginal fluid dyeing liquor
Preparation method is prepared.
A kind of colouring method of vaginal fluid dyeing liquor, wherein, vaginal fluid is contaminated using humidity strip decoration method
Color, the humidity strip decoration method are：Vaginal fluid through elution is added drop-wise on glass slide or in sample tube, later will
Dyeing liquor as described above is uniformly mixed with vaginal fluid, and the dyeing liquor is made to dye vaginal fluid.
Advantageous effect：The present invention using blue color basic dye and rosamines due to being mixed to prepare dyeing liquor so that
Dyeing liquor obtained has the coloring of general dyeing liquor, and can greatly shorten dyeing time, makes usually at 5 ~ 30 minutes
The dyeing time of interior completion shorten to 20s, greatly improves staining efficiency, accelerates the screening speed of clinical sample.
Description of the drawings
Fig. 1 is the coloring figure of conventional method.
Fig. 2 ~ Fig. 9 is the coloring figure of the method for the present invention.
Specific embodiment
The present invention provides a kind of vaginal fluid dyeing liquor and preparation method thereof and colouring method, to make the mesh of the present invention
, technical solution and effect it is clearer, clear and definite, the present invention is described in more detail below.It should be appreciated that described herein
Specific embodiment be only used to explain the present invention, be not intended to limit the present invention.
The present invention provides a kind of preparation method of vaginal fluid dyeing liquor, including step：
A, by weight, 1 ~ 3 part of blue color basic dye, 6 ~ 8 parts of rosamines, 1 ~ 3 part of buffer salt are weighed respectively,
8 ~ 12 parts of alcoholic solvent, 75 ~ 85 parts of deionized water；
B, by the above-mentioned blue color basic dye weighed, rosamines, buffer salt, alcoholic solvent and deionized water 22 ~
It is uniformly mixed, and stands overnight at 28 DEG C；
C, 0.02 ~ 0.5 part of alkaline matter, membrane protective agent 0.01 ~ 0.5 are added in the mixed liquor after standing overnight
Part, 0.01 ~ 0.5 part of dyeing promoter, 0.002 ~ 0.2 part of surfactant, 0.01 ~ 0.5 part of osmotic pressure regulator, it is finally quiet again
It puts overnight, obtains dyeing liquor.
Further, by weight, 1 part of blue color basic dye, 8 parts of rosamines, buffer salt 1 are weighed respectively
Part, 10 parts of alcoholic solvent, 80 parts of deionized water, to prepare vaginal fluid dyeing liquor.
Further, the blue color basic dye can be in methylenum careuleum, azure II, methyl green, crystal violet, toluidine blue
It is one or more.
Further, the rosamines can be dimethyl diaminophenazine chloride, safranine T, eosin Y, one kind in methyl red or more
Kind.
The main of the present invention thes improvement is that, described comprising blue color basic dye and rosamines in dyeing liquor
Blue color basic dye and rosamines are basic dye, the basic dye can abundant infiltrating cells, in secretion
The nucleus and cytoplasm of various types of cells can be dyed, and various types of cells in secretion can be distinguished.
Further, the buffer salt can be MES, borate, phosphate, Tris-HCl, Bistris-HCl,
It is one or more in HEPES, PIPES.And a concentration of 2 ~ 500mmol/L of the buffer salt, buffer salt pH are 5.0~7.8.
Further, the alcoholic solvent can be one kind in absolute ethyl alcohol, methanol, ethylene glycol.
Further, the alkaline matter can be natrium carbonicum calcinatum, sodium bicarbonate, potassium acetate, sodium acetate, calcium acetate,
It is one or more in borax.
Further, the membrane protective agent can be trehalose, sucrose, mannitol, one kind in glycine or more
Kind.
Further, the dyeing promoter can be cytase, mucolytic agent bromhexine, ambroxol, acetyl
It is one or more in cysteine.
Further, the surfactant can be alkyl phenol polyoxyethylene ether, high-carbon fatty alcohol polyoxyethylene ether, fat
Fat acid polyoxyethylene ester, fatty acid methyl ester ethoxylate, the ethylene oxide adduct of polypropylene glycol, sorbitan ester, sugarcane
It is one or more in sugar ester.
Further, the osmotic pressure regulator can be one in glycerine, PVP, glucose, fructose, polyglucose
Kind is a variety of.
Based on above-mentioned preparation method, the present invention also provides a kind of vaginal fluid dyeing liquors, use as above any described
The preparation method of vaginal fluid dyeing liquor be prepared.The present invention is due to containing blue color basic dye and red in dyeing liquor
Color basic dye, the basic dye easily with negatively charged chromatin with ionic bond or Hydrogenbond so as to adhere to chromatin
Corresponding color so that dyeing liquor has the coloring of general dyeing liquor, and can greatly shorten dyeing time, make to lead to
The dyeing time often completed in 5 ~ 30 minutes shorten to 20s, greatly improves staining efficiency, accelerates the screening of clinical sample
Speed.
Based on above-mentioned vaginal fluid dyeing liquor, the present invention also provides a kind of colouring method of vaginal fluid dyeing liquor,
The present invention dyes vaginal fluid above-mentioned dyeing liquor using humidity strip decoration method, and the humidity strip decoration method is：It will be through washing
The vaginal fluid of de- liquid elution is added drop-wise on glass slide or in sample tube, later by dyeing liquor as described above and vaginal fluid
It is uniformly mixed, the dyeing liquor is made to dye vaginal fluid.
Dyeing liquor prepared by the present invention dyes vaginal fluid using humidity strip decoration method, is substantially shorter dyeing
Time, the abundant infiltrating cells of basic dye dye the nucleus and cytoplasm of the various types of cells in secretion, and to dividing
Various types of cells distinguishes in secretion.In addition, dyeing time of the present invention can be 10 ~ 240s, can be specifically accustomed to according to operator
And sample to be tested number is automatically adjusted, coloring is not changed and is changed at any time.In addition, the dyeing side of the present invention
Method is the quick humidity strip decoration method of a step, and cross contamination caused by substep dyeing can be avoided possible simplifies operating procedure, makes dyeing
Process is easier；The present invention dyeing after image clearly, be easily recognized, can clearly identify surface layer squamous cell, in
The ingredients such as layer epithelial cell, bottom epithelial cell, clues cell, leucocyte, red blood cell, Candida albicans, trichomonad, suitable for big
Measure the Clinical screening of sample.
The present invention is described in further detail below by way of specific embodiment：
Embodiment 1：
Electronic balance weighs dimethyl diaminophenazine chloride 1g, methylenum careuleum 0.125g, glycerine 0.1mL, SDS 0.01g, 10mL absolute ethyl alcohol,
90mL deionized waters, dissolving dye, avoid light place 24~48 hours, filtering are spare.
Embodiment 2：
Electronic balance weighs dimethyl diaminophenazine chloride 1g, methylenum careuleum 0.1g, azure II 0.025g, glycerine 0.1mL, SDS 0.01g,
10mL absolute ethyl alcohols, 90mL MES buffer solutions, are protected from light dissolving dye 24~48 hours, and filtering is spare.
Embodiment 3：
Electronic balance weighs dimethyl diaminophenazine chloride 1g, methylenum careuleum 0.125g, glycerine 0.1mL, PVP 0.1g, SDS 0.01g, 10mL without
Water-ethanol, 90ml Bistris buffer solutions, is protected from light dissolving dye 24~48 hours, and filtering is spare.
Fig. 1 is the coloring figure of conventional coloring method；Fig. 2 ~ Fig. 9 is the coloring figure of colouring method of the present invention.Knot
Fruit analysis is found：Compared with conventional coloring method, the image after present invention dyeing is apparent, is easily recognized, can clearly identify
The ingredients such as epithelial cell, clues cell, leucocyte, red blood cell, Candida albicans, trichomonad.Therefore, coloring of the invention is excellent
In traditional dyeing effect.
In conclusion vaginal fluid dyeing liquor provided by the invention and its colouring method, the present invention is due to using blue
Basic dye and rosamines are mixed to prepare dyeing liquor so that the dyeing that dyeing liquor obtained has general dyeing liquor is imitated
Fruit, and dyeing time can be greatly shortened, the dyeing time usually completed in 5 ~ 30 minutes is made to shorten to 20s, is greatly improved
Staining efficiency accelerates the screening speed of clinical sample.In addition, using the image clearly after dyeing liquor of the present invention dyeing, easily
In identification, surface layer squamous cell can be clearly identified, middle level epithelial cell, bottom epithelial cell, clues cell is white thin
The ingredients such as born of the same parents, red blood cell, Candida albicans, trichomonad, suitable for the Clinical screening of great amount of samples.
It should be understood that the application of the present invention is not limited to the above, it for those of ordinary skills, can
To be improved or converted according to the above description, all these modifications and variations should all belong to the guarantor of appended claims of the present invention
Protect range.
Claims (8)
1. a kind of preparation method of vaginal fluid dyeing liquor, which is characterized in that including step：
A, by weight, 1 ~ 3 part of blue color basic dye, 6 ~ 8 parts of rosamines, 1 ~ 3 part of buffer salt are weighed respectively, and alcohol is molten
8 ~ 12 parts of agent, 75 ~ 85 parts of deionized water；
The blue color basic dye is methylenum careuleum, one or more in azure II, methyl green, crystal violet, toluidine blue；
The rosamines are dimethyl diaminophenazine chloride, one or more in safranine T, methyl red；
B, by the above-mentioned blue color basic dye weighed, rosamines, buffer salt, alcoholic solvent and deionized water are at 22 ~ 28 DEG C
It is lower to be uniformly mixed, and stand overnight；
C, 0.02 ~ 0.5 part of alkaline matter, 0.01 ~ 0.5 part of membrane protective agent, dye are added in the mixed liquor after standing overnight
0.01 ~ 0.5 part of color accelerating agent, 0.002 ~ 0.2 part of surfactant, 0.01 ~ 0.5 part of osmotic pressure regulator, finally stood again
Night obtains dyeing liquor；
Image clearly after being dyed using the dyeing liquor, is easily recognized, can clearly identify surface layer squamous cell, middle level
Epithelial cell, bottom epithelial cell, clues cell, leucocyte, red blood cell, Candida albicans, trichomonad, suitable for the clinic of sample
Screening.
2. the preparation method of vaginal fluid dyeing liquor according to claim 1, which is characterized in that the alkaline matter is
It is one or more in natrium carbonicum calcinatum, sodium bicarbonate, potassium acetate, sodium acetate, calcium acetate, borax.
3. the preparation method of vaginal fluid dyeing liquor according to claim 1, which is characterized in that the membrane protective
Agent is trehalose, one or more in sucrose, mannitol, glycine.
4. the preparation method of vaginal fluid dyeing liquor according to claim 1, which is characterized in that the dyeing promoter
It is one or more in cytase, mucolytic agent bromhexine, ambroxol, acetylcysteine.
5. the preparation method of vaginal fluid dyeing liquor according to claim 1, which is characterized in that the surfactant
For alkyl phenol polyoxyethylene ether, high-carbon fatty alcohol polyoxyethylene ether, polyoxyethylene carboxylate, fatty acid methyl ester ethoxylation
It is one or more in object, the ethylene oxide adduct of polypropylene glycol, sorbitan ester, sucrose ester.
6. the preparation method of vaginal fluid dyeing liquor according to claim 1, which is characterized in that the osmotic pressure is adjusted
Agent is glycerine, one or more in PVP, glucose, fructose, polyglucose.
7. a kind of vaginal fluid dyeing liquor, which is characterized in that using the vaginal fluid dye as described in claim 1 ~ 6 is any
The preparation method of color liquid is prepared.
8. a kind of colouring method of vaginal fluid dyeing liquor, which is characterized in that using humidity strip decoration method to vaginal fluid into
Row dyeing, the humidity strip decoration method are：Vaginal fluid through elution is added drop-wise on glass slide or in sample tube, it
Dyeing liquor as claimed in claim 7 with vaginal fluid is uniformly mixed afterwards, the dyeing liquor is made to contaminate vaginal fluid
Color.
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| CN112213172A (en) * | 2020-09-16 | 2021-01-12 | 迪瑞医疗科技股份有限公司 | Stable vaginal secretion visible component staining solution and preparation method thereof |
| CN112284864A (en) * | 2020-10-21 | 2021-01-29 | 桂林优利特医疗电子有限公司 | Preparation method of vaginal secretion staining solution and staining method |
| CN112525652A (en) * | 2020-11-13 | 2021-03-19 | 广州翰德泽信医药科技有限公司 | Stable and easily-preserved vaginal secretion microbial cell fluorescence detection dye solution |
| CN114441271B (en) * | 2021-12-27 | 2023-06-27 | 桂林优利特医疗电子有限公司 | Novel dyeing liquid preparation and dyeing method |
| CN114923757A (en) * | 2022-04-14 | 2022-08-19 | 北京泰格科信生物科技有限公司 | Staining fluid for visible components of vaginal secretion and preparation method thereof |
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