CN114813289A - Vaginal secretion fluorescent staining solution and staining method thereof - Google Patents
Vaginal secretion fluorescent staining solution and staining method thereof Download PDFInfo
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- CN114813289A CN114813289A CN202210438202.9A CN202210438202A CN114813289A CN 114813289 A CN114813289 A CN 114813289A CN 202210438202 A CN202210438202 A CN 202210438202A CN 114813289 A CN114813289 A CN 114813289A
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- 210000003756 cervix mucus Anatomy 0.000 title claims abstract description 54
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 53
- 239000000243 solution Substances 0.000 title claims abstract description 53
- 238000007447 staining method Methods 0.000 title abstract description 14
- 239000000523 sample Substances 0.000 claims abstract description 66
- 238000010186 staining Methods 0.000 claims abstract description 24
- 239000012470 diluted sample Substances 0.000 claims abstract description 21
- 239000006059 cover glass Substances 0.000 claims abstract description 17
- 239000011521 glass Substances 0.000 claims abstract description 17
- 239000011265 semifinished product Substances 0.000 claims abstract description 10
- 239000002250 absorbent Substances 0.000 claims abstract description 8
- 230000002745 absorbent Effects 0.000 claims abstract description 8
- 239000002504 physiological saline solution Substances 0.000 claims abstract description 7
- 230000028327 secretion Effects 0.000 claims abstract description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical group OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 30
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical group [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 30
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 20
- 108020004707 nucleic acids Proteins 0.000 claims description 20
- 102000039446 nucleic acids Human genes 0.000 claims description 20
- 150000007523 nucleic acids Chemical class 0.000 claims description 20
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 18
- 108090001090 Lectins Proteins 0.000 claims description 15
- 102000004856 Lectins Human genes 0.000 claims description 15
- 239000002523 lectin Substances 0.000 claims description 15
- 239000000980 acid dye Substances 0.000 claims description 14
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 claims description 14
- 239000007853 buffer solution Substances 0.000 claims description 12
- 239000003906 humectant Substances 0.000 claims description 12
- 229910019142 PO4 Inorganic materials 0.000 claims description 10
- 235000010323 ascorbic acid Nutrition 0.000 claims description 10
- 229960005070 ascorbic acid Drugs 0.000 claims description 10
- 239000011668 ascorbic acid Substances 0.000 claims description 10
- 239000000022 bacteriostatic agent Substances 0.000 claims description 10
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 10
- 239000010452 phosphate Substances 0.000 claims description 10
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims description 10
- 235000010288 sodium nitrite Nutrition 0.000 claims description 10
- ACOJCCLIDPZYJC-UHFFFAOYSA-M thiazole orange Chemical compound CC1=CC=C(S([O-])(=O)=O)C=C1.C1=CC=C2C(C=C3N(C4=CC=CC=C4S3)C)=CC=[N+](C)C2=C1 ACOJCCLIDPZYJC-UHFFFAOYSA-M 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 238000004043 dyeing Methods 0.000 claims description 9
- 239000000975 dye Substances 0.000 claims description 7
- 235000010518 Canavalia gladiata Nutrition 0.000 claims description 6
- 101000582927 Photorhabdus laumondii subsp. laumondii (strain DSM 15139 / CIP 105565 / TT01) Lectin A Proteins 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 238000010791 quenching Methods 0.000 claims description 6
- 206010046901 vaginal discharge Diseases 0.000 claims description 6
- 235000010520 Canavalia ensiformis Nutrition 0.000 claims description 4
- 244000045232 Canavalia ensiformis Species 0.000 claims description 3
- 240000003049 Canavalia gladiata Species 0.000 claims description 3
- 108010062580 Concanavalin A Proteins 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 101710186708 Agglutinin Proteins 0.000 claims description 2
- 101710146024 Horcolin Proteins 0.000 claims description 2
- 101710189395 Lectin Proteins 0.000 claims description 2
- 101710179758 Mannose-specific lectin Proteins 0.000 claims description 2
- 101710150763 Mannose-specific lectin 1 Proteins 0.000 claims description 2
- 101710150745 Mannose-specific lectin 2 Proteins 0.000 claims description 2
- 239000000910 agglutinin Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 244000005700 microbiome Species 0.000 abstract description 5
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 239000000047 product Substances 0.000 abstract description 3
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 102000003886 Glycoproteins Human genes 0.000 description 4
- 108090000288 Glycoproteins Proteins 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000012757 fluorescence staining Methods 0.000 description 2
- 239000000138 intercalating agent Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 241000220451 Canavalia Species 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000224526 Trichomonas Species 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
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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
- G01N1/2806—Means for preparing replicas of specimens, e.g. for microscopal 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
- G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
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Abstract
The invention relates to the technical field of medical reagents, in particular to a vaginal secretion fluorescent staining solution and a staining method thereof, which comprises the steps of preparing the vaginal secretion fluorescent staining solution; adding 0.5ml of physiological saline into the secretion sample, and uniformly mixing to obtain a diluted sample; dripping the diluted sample on a glass slide and baking the diluted sample to obtain a baked sample; adding a vaginal secretion fluorescent staining solution into the oven-dried sample for staining to obtain a semi-finished stained sample; covering the semi-finished product on the stained sample by using the cover glass; the fluorescent staining solution for vaginal secretions between the glass slide and the cover glass is sucked out by using the absorbent paper, and the staining method can clearly present the forms of cells and microorganisms, is more beneficial to the examination of doctors, obtains a finished stained sample product, and solves the problem that the existing wet staining method is easy to cause missed examination.
Description
Technical Field
The invention relates to the technical field of medical reagents, in particular to a vaginal secretion fluorescent staining solution and a staining method thereof.
Background
The vaginal secretion adopts a staining method to improve the detection rate of the causative factors.
At present, the fluorescence staining of vaginal secretion is wet staining, and by adopting a sample and a fluorescence staining solution and adding physiological saline or a diluent, visible components in the sample are suspended in the liquid, so that the focal length needs to be adjusted frequently, and the omission of pathogenic microorganisms is easily caused due to personal operation.
Disclosure of Invention
The invention aims to provide a vaginal secretion fluorescent staining solution and a staining method thereof, and aims to solve the problem that the existing wet staining method is easy to cause missed detection.
In order to achieve the above object, in a first aspect, the present invention provides a method for staining vaginal secretion with a fluorescent staining solution, comprising the following steps:
preparing a fluorescent staining solution for vaginal secretion;
adding 0.5ml of physiological saline into the secretion sample, and uniformly mixing to obtain a diluted sample;
dripping the diluted sample on a glass slide and baking the diluted sample to obtain a baked sample;
adding the vaginal secretion fluorescent staining solution to the baked sample for staining to obtain a stained sample semi-finished product;
covering a cover glass on the dyeing sample to obtain a semi-finished product;
and (3) sucking out the vaginal secretion fluorescent staining solution between the glass slide and the cover glass by using absorbent paper to obtain a finished stained sample.
The specific mode for preparing the vaginal secretion fluorescent staining solution is as follows:
uniformly mixing lectin, fluorescein, nucleic acid dye, anti-quenching agent, buffer solution, bacteriostatic agent, humectant, water and alkaline regulator at 25 +/-3 ℃ to obtain the vaginal secretion fluorescent staining solution.
Wherein, the lectin is sword bean lectin A, fluorescein is tetraethyl rhodamine, the nucleic acid dyestuff is thiazole orange, anti-quencher is ascorbic acid, buffer solution is the phosphate, the bacteriostat is for being sodium nitrite, the humectant is ethylene glycol, alkaline regulator is sodium hydroxide.
Wherein the baking temperature of the diluted sample is 42-70 ℃.
The method comprises the following steps of (1) adding the vaginal secretion fluorescent staining solution to the baked sample for staining, wherein the specific mode of obtaining a semi-finished stained sample is as follows:
and adding the vaginal secretion fluorescent staining solution to the baked sample for staining for 10-15 seconds to obtain a semi-finished stained sample.
Wherein, after the step of sucking out the fluorescent staining solution of vaginal secretion between the glass slide and the cover glass by using absorbent paper to obtain a finished stained sample, the method further comprises the following steps:
placing the glass slide, the finished dyed sample and the cover glass under a microscope, and selecting blue light with wavelength of 488nm as exciting light to excite the finished dyed sample to obtain a fluorescent sample;
observing the fluorescent sample through the microscope.
In a second aspect, the invention provides a preparation of a vaginal secretion fluorescent staining solution, which comprises lectin, fluorescein, a nucleic acid dye, an anti-quencher, a buffer solution, a bacteriostatic agent, a humectant, water and an alkalinity regulator;
the lectin is Canavalia gladiata lectin A, the fluorescein is tetraethylrhodamine, the nucleic acid dye is thiazole orange, the anti-quenching agent is ascorbic acid, the buffer solution is phosphate, the bacteriostatic agent is sodium nitrite, the humectant is ethylene glycol, and the alkaline regulator is sodium hydroxide;
the food additive comprises, by weight, 1-5 parts of jack bean agglutinin A, 1-5 parts of tetraethyl rhodamine, 0.3-3 parts of thiazole orange, 1-5 parts of ascorbic acid, 0.1-3 parts of phosphate, 0.1-3 parts of sodium nitrite, 0.5-2.8 parts of ethylene glycol, 0.1-0.85 part of water and 0.1-0.85 part of sodium hydroxide.
The invention relates to a method for staining vaginal secretion by using fluorescent staining solution, which comprises the steps of preparing the vaginal secretion fluorescent staining solution; adding 0.5ml of physiological saline into the secretion sample, and uniformly mixing to obtain a diluted sample; dripping the diluted sample on a glass slide and baking the diluted sample to obtain a baked sample; adding the vaginal secretion fluorescent staining solution to the baked sample for staining to obtain a stained sample semi-finished product; covering a cover glass on the dyeing sample to obtain a semi-finished product; the fluorescent staining solution for vaginal secretions between the glass slide and the cover glass is sucked out by using absorbent paper, and the staining method can clearly present the forms of cells and microorganisms, is more beneficial to the examination of doctors, obtains a finished stained sample product, and solves the problem that the existing wet staining method easily causes missed examination.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a flow chart of a method for staining vaginal secretion with a fluorescent staining solution provided by the invention.
FIG. 2 is a graph of the staining effect of stain 1 and stain 2.
FIG. 3 is a graph showing the staining effect of the fluorescent staining solution for vaginal secretion of the present invention.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative and intended to explain the present invention and should not be construed as limiting the present invention.
Referring to fig. 1 to 3, in a first aspect, the present invention provides a method for staining vaginal secretion with fluorescent staining solution, comprising the following steps:
s1, preparing a vaginal secretion fluorescent staining solution;
specifically, the lectin, the fluorescein, the nucleic acid dye, the anti-quencher, the buffer solution, the bacteriostatic agent, the humectant, the water and the alkaline regulator are uniformly mixed at 25 +/-3 ℃ to obtain the vaginal secretion fluorescent staining solution. The lectin is sword bean lectin A, fluorescein is tetraethyl rhodamine, the nucleic acid dyestuff is thiazole orange, anti quencher is ascorbic acid, buffer solution is the phosphate, the bacteriostat is for being sodium nitrite, the humectant is ethylene glycol, alkaline regulator is sodium hydroxide. The composition comprises, by weight, 1-5 parts of concanavalin A, 1-5 parts of tetraethylrhodamine, 0.3-3 parts of thiazole orange, 1-5 parts of ascorbic acid, 0.1-3 parts of phosphate, 0.1-3 parts of sodium nitrite, 0.5-2.8 parts of ethylene glycol, 0.1-0.85 part of water and 0.1-0.85 part of sodium hydroxide.
S2, adding 0.5ml of physiological saline into the secretion sample, and mixing uniformly to obtain a diluted sample;
s3, dripping the diluted sample on a glass slide and baking the diluted sample to obtain a baked sample;
specifically, the baking temperature of the diluted sample is 42-70 ℃.
S4, adding the vaginal secretion fluorescent staining solution to the baked sample for staining to obtain a semi-finished stained sample;
specifically, the vaginal secretion fluorescent staining solution is added to the oven-dried sample for staining for 10-15 seconds to obtain a semi-finished stained sample.
Dyeing principle: the lectin is a non-immunogenic protein, has a molecular weight of 11000 to 335000, has polyvalent binding ability, and can be combined with various labels. Can be used as a specific probe for histochemistry, and the binding site of the probe is shown under an optical microscope and an electron microscope. Therefore, the method is widely used for researching the property and distribution of the glycoprotein and the change of the glycoprotein in the normal cell renewal process. Can be used as a specific probe of tissue chemistry to be widely used for observing a sample under an optical microscope. Specific recognition of glycoproteins by the lectin and specific staining of nucleic acids by nucleic acid dyes. The nucleic acid dye belongs to a nucleic acid molecule intercalator, is easy to permeate a cell membrane and a micromolecule embedded with intracellular DNA, has a plane conjugated macrocyclic structure, is a typical DNA molecule intercalator, forms a stable complex with the DNA embedding, influences the DNA replication and destroys the normal hereditary physiological phenomenon. And simultaneously, the principle of fluorescence labeling of cells and microorganisms is also provided.
S5, covering a cover glass on the dyeing sample to obtain a semi-finished product;
s6, sucking out the vaginal secretion fluorescent staining solution between the glass slide and the cover glass by using absorbent paper to obtain a finished stained sample.
S7, placing the glass slide, the finished dyed sample and the cover glass under a microscope, and selecting blue light with wavelength of 488nm as exciting light to excite the finished dyed sample to obtain a fluorescent sample;
s8 observing the fluorescent sample through the microscope.
Has the advantages that:
simple preparation of fluorescent staining solution;
the problem that the wet dyeing is easy to miss detection due to different layers is solved;
the optimal nucleic acid dye is selected without the need to switch wavelengths.
The detection rate of pathogenic bacteria is better;
the invention relates to a method for staining vaginal secretion by using fluorescent staining solution, which comprises the steps of preparing the vaginal secretion fluorescent staining solution; adding 0.5ml of physiological saline into the secretion sample, and uniformly mixing to obtain a diluted sample; dripping the diluted sample on a glass slide and baking the diluted sample to obtain a baked sample; adding the vaginal secretion fluorescent staining solution to the baked sample for staining to obtain a stained sample semi-finished product; covering a cover glass on the dyeing sample to obtain a semi-finished product; the fluorescent staining solution for vaginal secretions between the glass slide and the cover glass is sucked out by using absorbent paper, and the staining method can clearly present the forms of cells and microorganisms, is more beneficial to the examination of doctors, obtains a finished stained sample product, and solves the problem that the existing wet staining method easily causes missed examination.
The detection results (120 cases) of several important components in the invention under various preparation methods with different parts are shown in table 1:
TABLE 1
The results of staining with 2 dyes purchased and the staining method of the present invention on trichomonas, fungi, clue cells, and pathogenic infectious microbes are shown in table 2:
TABLE 2
As shown in figures 2 to 3, the effect of the dye 2 is better than that of the dye 1, but the effect of the dye 2 is not as good as that of the fluorescent staining solution for vaginal secretion in the patent of the invention.
In a second aspect, the invention provides a vaginal secretion fluorescent staining solution, which comprises lectin, fluorescein, nucleic acid dye, an anti-quencher, a buffer solution, a bacteriostatic agent, a humectant, water and an alkalinity regulator;
the lectin is Canavalia gladiata lectin A, the fluorescein is tetraethylrhodamine, the nucleic acid dye is thiazole orange, the anti-quenching agent is ascorbic acid, the buffer solution is phosphate, the bacteriostatic agent is sodium nitrite, the humectant is ethylene glycol, and the alkaline regulator is sodium hydroxide;
the composition comprises, by weight, 1-5 parts of concanavalin A, 1-5 parts of tetraethylrhodamine, 0.3-3 parts of thiazole orange, 1-5 parts of ascorbic acid, 0.1-3 parts of phosphate, 0.1-3 parts of sodium nitrite, 0.5-2.8 parts of ethylene glycol, 0.1-0.85 part of water and 0.1-0.85 part of sodium hydroxide.
Specifically, the microbial fluorescent staining solution provided by the invention utilizes specific recognition of the lectin on glycoprotein and specific staining of nucleic acid by nucleic acid dye, labels the fluorescein through the lectin, and mixes the fluorescein with the nucleic acid dye, the bacteriostatic agent and the quencher according to a scientific and reasonable ratio. So as to realize the staining of cells and microorganisms.
While the present invention has been described with reference to preferred embodiments and methods for staining vaginal discharge, it will be understood by those skilled in the art that the present invention is not limited thereto, and that the present invention is capable of being practiced in the full scope of the invention.
Claims (7)
1. A method for staining vaginal secretion by using a fluorescent staining solution is characterized by comprising the following steps:
preparing a fluorescent staining solution for vaginal secretion;
adding 0.5ml of physiological saline into the secretion sample, and uniformly mixing to obtain a diluted sample;
dripping the diluted sample on a glass slide and baking the diluted sample to obtain a baked sample;
adding the vaginal secretion fluorescent staining solution to the baked sample for staining to obtain a stained sample semi-finished product;
covering a cover glass on the dyeing sample to obtain a semi-finished product;
and (3) sucking out the vaginal secretion fluorescent staining solution between the glass slide and the cover glass by using absorbent paper to obtain a finished stained sample.
2. The method of claim 1, wherein the vaginal discharge is stained with a fluorescent staining solution,
the specific mode for preparing the vaginal secretion fluorescent staining solution is as follows:
uniformly mixing lectin, fluorescein, nucleic acid dye, anti-quenching agent, buffer solution, bacteriostatic agent, humectant, water and alkaline regulator at 25 +/-3 ℃ to obtain the vaginal secretion fluorescent staining solution.
3. The method of claim 2, wherein the vaginal discharge is stained with a fluorescent staining solution,
the lectin is sword bean lectin A, fluorescein is tetraethyl rhodamine, the nucleic acid dyestuff is thiazole orange, anti quencher is ascorbic acid, buffer solution is the phosphate, the bacteriostat is for being sodium nitrite, the humectant is ethylene glycol, alkaline regulator is sodium hydroxide.
4. The method of claim 1, wherein the vaginal discharge is stained with a fluorescent staining solution,
the baking temperature of the diluted sample is 42-70 ℃.
5. The method of claim 1, wherein the vaginal discharge is stained with a fluorescent staining solution,
the specific method for obtaining the semi-finished dyed sample by adding the vaginal secretion fluorescent dyeing liquid to the oven-dried sample for dyeing comprises the following steps:
and adding the vaginal secretion fluorescent staining solution to the baked sample for staining for 10-15 seconds to obtain a semi-finished stained sample.
6. The method of claim 1, wherein the vaginal discharge is stained with a fluorescent staining solution,
after the step of sucking out the fluorescent staining solution of vaginal secretion between the glass slide and the cover glass by using absorbent paper to obtain a finished stained sample, the method further comprises the following steps:
placing the glass slide, the finished dyed sample and the cover glass under a microscope, and selecting blue light with wavelength of 488nm as exciting light to excite the finished dyed sample to obtain a fluorescent sample;
observing the fluorescent sample through the microscope.
7. A fluorescent staining solution for vaginal secretion, which is characterized in that,
comprises agglutinin, fluorescein, nucleic acid dye, anti-quenching agent, buffer solution, bacteriostatic agent, humectant, water and alkalinity regulator;
the lectin is Canavalia gladiata lectin A, the fluorescein is tetraethylrhodamine, the nucleic acid dye is thiazole orange, the anti-quenching agent is ascorbic acid, the buffer solution is phosphate, the bacteriostatic agent is sodium nitrite, the humectant is ethylene glycol, and the alkaline regulator is sodium hydroxide;
the composition comprises, by weight, 1-5 parts of concanavalin A, 1-5 parts of tetraethylrhodamine, 0.3-3 parts of thiazole orange, 1-5 parts of ascorbic acid, 0.1-3 parts of phosphate, 0.1-3 parts of sodium nitrite, 0.5-2.8 parts of ethylene glycol, 0.1-0.85 part of water and 0.1-0.85 part of sodium hydroxide.
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| CN202210438202.9A CN114813289A (en) | 2022-04-20 | 2022-04-20 | Vaginal secretion fluorescent staining solution and staining method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115683790A (en) * | 2022-11-04 | 2023-02-03 | 深圳市第二人民医院(深圳市转化医学研究院) | Fluorescent staining agent, preparation method thereof and staining method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115683790A (en) * | 2022-11-04 | 2023-02-03 | 深圳市第二人民医院(深圳市转化医学研究院) | Fluorescent staining agent, preparation method thereof and staining method |
| CN115683790B (en) * | 2022-11-04 | 2024-01-19 | 深圳市第二人民医院(深圳市转化医学研究院) | Fluorescent dye, preparation method thereof and dyeing method |
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