CN105067412A - Vaginal secretion staining fluid, and preparation method and staining method thereof - Google Patents
Vaginal secretion staining fluid, and preparation method and staining method thereof Download PDFInfo
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- CN105067412A CN105067412A CN201510484688.XA CN201510484688A CN105067412A CN 105067412 A CN105067412 A CN 105067412A CN 201510484688 A CN201510484688 A CN 201510484688A CN 105067412 A CN105067412 A CN 105067412A
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Abstract
The invention discloses a vaginal secretion staining fluid, and a preparation method and a staining method thereof. According to the preparation method, a blue basic dyestuff and a red basic dyestuff are mixed so as to obtain the vaginal secretion staining fluid, so that the vaginal secretion staining fluid possesses staining effects of universal staining fluid, staining time can be shortened greatly, staining time is shortened from general 5-30min to 20s, staining efficiency is increased greatly, and screening speed of clinical samples is accelerated. And in addition, imagines obtained via staining with the vaginal secretion staining fluid are clear, and are convenient for identification; surface layer squamous epithelial cells, middle layer epithelial cells, bottom layer epithelial cells, clue cells, leukocytes, erythrocytes, monilia albicans, and trichomonad can be identified clearly; and the vaginal secretion staining fluid is suitable for large-scale sample clinical screening.
Description
Technical field
The present invention relates to cell dyeing technical field, particularly relate to a kind of vaginal fluid dyeing liquor and preparation method thereof and colouring method.
Background technology
Endonuclear chromatin is DNA (deoxyribonucleic acid) (DNA) mainly, in DNA double helical structure, the phosphate on two chains is toward the outer side, electronegative, easy and positively charged basic-dyeable fibre with ionic link or Hydrogenbond, thus makes chromatin adhere to corresponding color.Nucleus is chromatinic storage place, and chromatin is fine and close, then nucleus color depth; Chromatin loosens, then nucleus is painted shallow.In secretion tenuigenin, albumen iso-electric point is 3 ~ 6, and as pH>PI, carboxylic group free in albumen increases, and can be combined, thus make tenuigenin adhere to corresponding color with positively charged basic-dyeable fibre.Tenuigenin institute is electronegative many, then tenuigenin dyeing is relatively dark; Tenuigenin institute is electronegative few, then tenuigenin dyeing is relatively shallow.And due to the cell in secretion and asynchronous propagation, the cell color degree be therefore in the different cell cycle there are differences.The cell being in growth period and division stage due to DNA and protein synthesis vigorous and present nucleus and color depth, the painted relatively shallow phenomenon of tenuigenin.And the cell being in apoptosis period due to permeability of cell membrane strengthen, in protein acidic amino acid site expose, chromatin dna segment degradation and that cell feulgen's stain is distinguished is not obvious.
At present, domestic application is vaginal fluid dyeing liquor mainly Rui Shi-Ji's nurse Sa dyeing liquor and gram staining liquid comparatively widely, but these two kinds of dyeing liquors all have the shortcoming such as complex operation step, dyeing time length, are unfavorable for that larger scale clinical is applied.
Therefore, prior art has yet to be improved and developed.
Summary of the invention
In view of above-mentioned the deficiencies in the prior art, the object of the present invention is to provide a kind of vaginal fluid dyeing liquor and preparation method thereof and colouring method, be intended to solve that existing dyeing liquor exists complex operation step, dyeing time is long, and be unfavorable for the problem that larger scale clinical is applied.
Technical scheme of the present invention is as follows:
A preparation method for vaginal fluid dyeing liquor, wherein, comprises step:
A, by weight, take blue color basic dye 1 ~ 3 part, rosamines 6 ~ 8 parts, buffer salt 1 ~ 3 part respectively, alcoholic solvent 8 ~ 12 parts, deionized water 75 ~ 85 parts;
B, by the above-mentioned blue color basic dye, rosamines, the buffer salt that take, alcoholic solvent and deionized water mix at 22 ~ 28 DEG C, and hold over night;
C, in the mixed liquor after hold over night, add alkaline matter 0.02 ~ 0.5 part, membrane protective agent 0.01 ~ 0.5 part, dyeing promoter 0.01 ~ 0.5 part, 0.002 ~ 0.2 part, surfactant, osmotic pressure regulator 0.01 ~ 0.5 part; last hold over night again, obtains dyeing liquor.
The preparation method of described vaginal fluid dyeing liquor, wherein, described blue color basic dye is one or more in methylenum careuleum, azure II, methyl green, methyl violet, toluidine blue.
The preparation method of described vaginal fluid dyeing liquor, wherein, described rosamines is one or more in dimethyl diaminophenazine chloride, safranine T, eosin W or W S, methyl red.
The preparation method of described vaginal fluid dyeing liquor, wherein, described alkaline matter is one or more in natrium carbonicum calcinatum, sodium bicarbonate, potassium acetate, sodium acetate, calcium acetate, borax.
The preparation method of described vaginal fluid dyeing liquor, wherein, described membrane protective agent is one or more in trehalose, sucrose, sweet mellow wine, glycocoll.
The preparation method of described vaginal fluid dyeing liquor, wherein, described dyeing promoter is one or more in cytase, mucolytic agent bromhexine, ambroxol, acetylcysteine.
The preparation method of described vaginal fluid dyeing liquor, wherein, described surfactant is one or more in APES, high-carbon fatty alcohol polyoxyethylene ether, polyoxyethylene carboxylate, fatty acid methyl ester ethoxylate, the ethylene oxide adduct of polypropylene glycol, sorbitan ester, sucrose ester.
The preparation method of described vaginal fluid dyeing liquor, wherein, described osmotic pressure regulator is one or more in glycerine, PVP, glucose, fructose, polyglucose.
A kind of vaginal fluid dyeing liquor, is characterized in that, adopts the preparation method of as above arbitrary described vaginal fluid dyeing liquor to be prepared from.
A kind of colouring method of vaginal fluid dyeing liquor, wherein, humidity strip decoration method is adopted to dye to vaginal fluid, described humidity strip decoration method is: be added drop-wise on microslide or in sample tube by the vaginal fluid through elution, afterwards dyeing liquor described above is mixed with vaginal fluid, described dyeing liquor is dyeed to vaginal fluid.
Beneficial effect: the present invention is owing to adopting blue color basic dye and the obtained dyeing liquor of rosamines mixing, the dyeing liquor obtained is made to have the Color of general dyeing liquor, and greatly can shorten dyeing time, the dyeing time usually completed in 5 ~ 30 minutes is made to shorten to 20s, greatly improve staining efficiency, accelerate the examination speed of clinical sample.
Accompanying drawing explanation
Fig. 1 is the Color figure of classic method.
Fig. 2 ~ Fig. 9 is the Color figure of the inventive method.
Embodiment
The invention provides a kind of vaginal fluid dyeing liquor and preparation method thereof and colouring method, for making object of the present invention, technical scheme and effect clearly, clearly, the present invention is described in more detail below.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
The invention provides a kind of preparation method of vaginal fluid dyeing liquor, it comprises step:
A, by weight, take blue color basic dye 1 ~ 3 part, rosamines 6 ~ 8 parts, buffer salt 1 ~ 3 part respectively, alcoholic solvent 8 ~ 12 parts, deionized water 75 ~ 85 parts;
B, by the above-mentioned blue color basic dye, rosamines, the buffer salt that take, alcoholic solvent and deionized water mix at 22 ~ 28 DEG C, and hold over night;
C, in the mixed liquor after hold over night, add alkaline matter 0.02 ~ 0.5 part, membrane protective agent 0.01 ~ 0.5 part, dyeing promoter 0.01 ~ 0.5 part, 0.002 ~ 0.2 part, surfactant, osmotic pressure regulator 0.01 ~ 0.5 part; last hold over night again, obtains dyeing liquor.
Further, by weight, take blue color basic dye 1 part, rosamines 8 parts, buffer salt 1 part respectively, alcoholic solvent 10 parts, deionized water 80 parts, to prepare vaginal fluid dyeing liquor.
Further, described blue color basic dye can be one or more in methylenum careuleum, azure II, methyl green, methyl violet, toluidine blue.
Further, described rosamines can be one or more in dimethyl diaminophenazine chloride, safranine T, eosin W or W S, methyl red.
Main improvements of the present invention are, blue color basic dye and rosamines is comprised in dyeing liquor, described blue color basic dye and rosamines are basic-dyeable fibre, described basic-dyeable fibre can abundant infiltrating cells, all can dye to the nucleus of the various types of cells in secretion and tenuigenin, and can distinguish various types of cells in secretion.
Further, described buffer salt can be one or more in MES, borate, phosphate, Tris-HCl, Bistris-HCl, HEPES, PIPES.And the concentration of described buffer salt is 2 ~ 500mmol/L, buffer salt pH is 5.0 ~ 7.8.
Further, described alcoholic solvent can be the one in absolute ethyl alcohol, methyl alcohol, ethylene glycol.
Further, described alkaline matter can be one or more in natrium carbonicum calcinatum, sodium bicarbonate, potassium acetate, sodium acetate, calcium acetate, borax.
Further, described membrane protective agent can be one or more in trehalose, sucrose, sweet mellow wine, glycocoll.
Further, described dyeing promoter can be one or more in cytase, mucolytic agent bromhexine, ambroxol, acetylcysteine.
Further, described surfactant can be one or more in APES, high-carbon fatty alcohol polyoxyethylene ether, polyoxyethylene carboxylate, fatty acid methyl ester ethoxylate, the ethylene oxide adduct of polypropylene glycol, sorbitan ester, sucrose ester.
Further, described osmotic pressure regulator can be one or more in glycerine, PVP, glucose, fructose, polyglucose.
Based on above-mentioned preparation method, the present invention also provides a kind of vaginal fluid dyeing liquor, and it adopts the preparation method of as above arbitrary described vaginal fluid dyeing liquor to be prepared from.The present invention is owing to containing blue color basic dye and rosamines in dyeing liquor, this basic-dyeable fibre easily makes chromatin adhere to corresponding color with ionic link or Hydrogenbond to electronegative chromatin, thus make dyeing liquor have the Color of general dyeing liquor, and greatly can shorten dyeing time, the dyeing time usually completed in 5 ~ 30 minutes is made to shorten to 20s, greatly improve staining efficiency, accelerate the examination speed of clinical sample.
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 adopts humidity strip decoration method to be dyeed to vaginal fluid by above-mentioned dyeing liquor, described humidity strip decoration method is: be added drop-wise on microslide or in sample tube by the vaginal fluid through elution, afterwards dyeing liquor described above is mixed with vaginal fluid, described dyeing liquor is dyeed to vaginal fluid.
The dyeing liquor adopting humidity strip decoration method the present invention to be prepared dyes to vaginal fluid, greatly can shorten dyeing time, the abundant infiltrating cells of basic-dyeable fibre, dyes to the nucleus of the various types of cells in secretion and tenuigenin, and distinguishes various types of cells in secretion.In addition, dyeing time of the present invention can be 10 ~ 240s, and specifically automatically can adjust according to operator's custom and sample to be tested number, Color does not change in time and changes.In addition, colouring method of the present invention is the quick humidity strip decoration method of a step, can avoid dyeing step by step the cross pollution that may cause, simplify operation steps, make dyeing course easier; Clear picture after the present invention's dyeing, be easy to identification, can the composition such as clear identification top layer squamous cell, middle level epithelial cell, bottom epithelial cell, clues cell, leucocyte, red blood cell, Candida albicans, trichomonad, be applicable to the Clinical screening of great amount of samples.
Below by way of specific embodiment, the present invention is described in further detail:
Embodiment 1:
Electronic balance takes dimethyl diaminophenazine chloride 1g, methylenum careuleum 0.125g, glycerine 0.1mL, SDS0.01g, 10mL absolute ethyl alcohol, 90mL deionized water, dissolving dye, and lucifuge is placed 24 ~ 48 hours, filters for subsequent use.
Embodiment 2:
Electronic balance takes dimethyl diaminophenazine chloride 1g, methylenum careuleum 0.1g, azure II 0.025g, glycerine 0.1mL, SDS0.01g, 10mL absolute ethyl alcohol, 90mLMES damping fluid, and lucifuge dissolving dye 24 ~ 48 hours, filters for subsequent use.
Embodiment 3:
Electronic balance takes dimethyl diaminophenazine chloride 1g, methylenum careuleum 0.125g, glycerine 0.1mL, PVP0.1g, SDS0.01g, 10mL absolute ethyl alcohol, 90mlBistris damping fluid, and lucifuge dissolving dye 24 ~ 48 hours, filters for subsequent use.
Fig. 1 is the Color figure of conventional coloring method; Fig. 2 ~ Fig. 9 is the Color figure of colouring method of the present invention.Interpretation of result finds: compared with conventional coloring method, and the image after the present invention's dyeing is more clear, is easy to identification, can the composition such as clear identification epithelial cell, clues cell, leucocyte, red blood cell, Candida albicans, trichomonad.Therefore, Color of the present invention is better than traditional dyeing effect.
In sum, vaginal fluid dyeing liquor provided by the invention and colouring method thereof, the present invention is owing to adopting blue color basic dye and the obtained dyeing liquor of rosamines mixing, the dyeing liquor obtained is made to have the Color of general dyeing liquor, and greatly can shorten dyeing time, make the dyeing time usually completed in 5 ~ 30 minutes shorten to 20s, greatly improve staining efficiency, accelerate the examination speed of clinical sample.In addition, the clear picture after adopting dyeing liquor of the present invention to dye, is easy to identification, can clear identification top layer squamous cell, middle level epithelial cell, bottom epithelial cell, clues cell, leucocyte, red blood cell, Candida albicans, the compositions such as trichomonad, are applicable to the Clinical screening of great amount of samples.
Should be understood that, application of the present invention is not limited to above-mentioned citing, for those of ordinary skills, can be improved according to the above description or convert, and all these improve and convert the protection domain that all should belong to claims of the present invention.
Claims (10)
1. a preparation method for vaginal fluid dyeing liquor, is characterized in that, comprises step:
A, by weight, take blue color basic dye 1 ~ 3 part, rosamines 6 ~ 8 parts, buffer salt 1 ~ 3 part respectively, alcoholic solvent 8 ~ 12 parts, deionized water 75 ~ 85 parts;
B, by the above-mentioned blue color basic dye, rosamines, the buffer salt that take, alcoholic solvent and deionized water mix at 22 ~ 28 DEG C, and hold over night;
C, in the mixed liquor after hold over night, add alkaline matter 0.02 ~ 0.5 part, membrane protective agent 0.01 ~ 0.5 part, dyeing promoter 0.01 ~ 0.5 part, 0.002 ~ 0.2 part, surfactant, osmotic pressure regulator 0.01 ~ 0.5 part; last hold over night again, obtains dyeing liquor.
2. the preparation method of vaginal fluid dyeing liquor according to claim 1, is characterized in that, described blue color basic dye is one or more in methylenum careuleum, azure II, methyl green, methyl violet, toluidine blue.
3. the preparation method of vaginal fluid dyeing liquor according to claim 1, is characterized in that, described rosamines is one or more in dimethyl diaminophenazine chloride, safranine T, eosin W or W S, methyl red.
4. the preparation method of vaginal fluid dyeing liquor according to claim 1, is characterized in that, described alkaline matter is one or more in natrium carbonicum calcinatum, sodium bicarbonate, potassium acetate, sodium acetate, calcium acetate, borax.
5. the preparation method of vaginal fluid dyeing liquor according to claim 1, is characterized in that, described membrane protective agent is one or more in trehalose, sucrose, sweet mellow wine, glycocoll.
6. the preparation method of vaginal fluid dyeing liquor according to claim 1, is characterized in that, described dyeing promoter is one or more in cytase, mucolytic agent bromhexine, ambroxol, acetylcysteine.
7. the preparation method of vaginal fluid dyeing liquor according to claim 1, it is characterized in that, described surfactant is one or more in APES, high-carbon fatty alcohol polyoxyethylene ether, polyoxyethylene carboxylate, fatty acid methyl ester ethoxylate, the ethylene oxide adduct of polypropylene glycol, sorbitan ester, sucrose ester.
8. the preparation method of vaginal fluid dyeing liquor according to claim 1, is characterized in that, described osmotic pressure regulator is one or more in glycerine, PVP, glucose, fructose, polyglucose.
9. a vaginal fluid dyeing liquor, is characterized in that, adopt as arbitrary in claim 1 ~ 8 as described in the preparation method of vaginal fluid dyeing liquor be prepared from.
10. the colouring method of a vaginal fluid dyeing liquor, it is characterized in that, humidity strip decoration method is adopted to dye to vaginal fluid, described humidity strip decoration method is: be added drop-wise on microslide or in sample tube by the vaginal fluid through elution, to mix with vaginal fluid by dyeing liquor as claimed in claim 9 afterwards, described dyeing liquor is dyeed to vaginal fluid.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107167356A (en) * | 2017-03-31 | 2017-09-15 | 迪瑞医疗科技股份有限公司 | In a kind of urine after cell dyeing quick colour-developing coloring agent and its application method |
| CN109946140A (en) * | 2019-04-13 | 2019-06-28 | 三门县人民医院 | Fungi dyeing liquor and fungi colouring method |
| CN110006724A (en) * | 2019-04-17 | 2019-07-12 | 郑州安图生物工程股份有限公司 | Trichomonad reagent is detected using Pasteur and Gram-staining process |
| CN112067409A (en) * | 2020-08-06 | 2020-12-11 | 佛山科学技术学院 | Improved H.E staining method for mouse bone marrow tissue section |
| 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 |
| CN112781961A (en) * | 2019-11-07 | 2021-05-11 | 北京望升伟业科技发展有限公司 | Rapid cell staining solution and application thereof |
| CN114441271A (en) * | 2021-12-27 | 2022-05-06 | 桂林优利特医疗电子有限公司 | Preparation of novel dyeing liquid 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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| CN107167356A (en) * | 2017-03-31 | 2017-09-15 | 迪瑞医疗科技股份有限公司 | In a kind of urine after cell dyeing quick colour-developing coloring agent and its application method |
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| CN109946140B (en) * | 2019-04-13 | 2021-05-11 | 三门县人民医院 | Fungus staining solution and fungus staining method |
| CN110006724A (en) * | 2019-04-17 | 2019-07-12 | 郑州安图生物工程股份有限公司 | Trichomonad reagent is detected using Pasteur and Gram-staining process |
| CN110006724B (en) * | 2019-04-17 | 2021-06-04 | 郑州安图生物工程股份有限公司 | Reagent for detecting trichomonas by adopting papanicolaou and gram staining method |
| CN112781961A (en) * | 2019-11-07 | 2021-05-11 | 北京望升伟业科技发展有限公司 | Rapid cell staining solution and application thereof |
| CN112067409A (en) * | 2020-08-06 | 2020-12-11 | 佛山科学技术学院 | Improved H.E staining method for mouse bone marrow tissue section |
| 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 |
| CN114441271A (en) * | 2021-12-27 | 2022-05-06 | 桂林优利特医疗电子有限公司 | Preparation of novel dyeing liquid and dyeing method |
| 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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