CN107300495A - A kind of colouring method of pyriform worm blood film compound stain solution and blood film - Google Patents
A kind of colouring method of pyriform worm blood film compound stain solution and blood film Download PDFInfo
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- CN107300495A CN107300495A CN201710450703.8A CN201710450703A CN107300495A CN 107300495 A CN107300495 A CN 107300495A CN 201710450703 A CN201710450703 A CN 201710450703A CN 107300495 A CN107300495 A CN 107300495A
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- 210000004369 blood Anatomy 0.000 title claims abstract description 53
- 239000008280 blood Substances 0.000 title claims abstract description 53
- 150000001875 compounds Chemical class 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000004040 coloring Methods 0.000 title claims abstract description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 39
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000012153 distilled water Substances 0.000 claims abstract description 7
- 235000011187 glycerol Nutrition 0.000 claims abstract description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 7
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 7
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000000049 pigment Substances 0.000 claims abstract description 7
- 238000002360 preparation method Methods 0.000 claims abstract description 6
- 239000000975 dye Substances 0.000 claims description 16
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 4
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 4
- 239000011574 phosphorus Substances 0.000 claims description 4
- 229910052698 phosphorus Inorganic materials 0.000 claims description 4
- 239000011734 sodium Substances 0.000 claims description 4
- 229910052708 sodium Inorganic materials 0.000 claims description 4
- 210000004027 cell Anatomy 0.000 claims description 3
- 238000009826 distribution Methods 0.000 claims description 3
- 210000003743 erythrocyte Anatomy 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 239000004570 mortar (masonry) Substances 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims 1
- 239000002131 composite material Substances 0.000 abstract description 2
- 230000000694 effects Effects 0.000 abstract description 2
- 239000000843 powder Substances 0.000 abstract 1
- 201000008680 babesiosis Diseases 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 241000223836 Babesia Species 0.000 description 2
- 241000283073 Equus caballus Species 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 241000223775 Babesia caballi Species 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 206010023126 Jaundice Diseases 0.000 description 1
- 208000030852 Parasitic disease Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 241000282849 Ruminantia Species 0.000 description 1
- 241000223777 Theileria Species 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 208000024891 symptom Diseases 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
- G01N1/2813—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)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to biological technical field, and in particular to a kind of pyriform worm blood film compound stain solution includes compound stain solution I and compound dye night II;Described compound stain solution I is made up of Switzerland powder 2g, Jim Sa pigment 1.7g, glycerine 50ml and methanol 1000ml, and compound stain solution II is made up of AMSP 2.3g, anhydrous potassium dihydrogenphosphate 0.3g and 1000ml distilled water.Colouring method comprises the steps:First, the preparation of blood film;2nd, blood film after-treatment;3rd, dyed in compound stain solution.The composite dye and colouring method of the present invention is preferable to pyriform worm Color in the case where being not required to by high-end instrument and equipment, and economical and practical, required time is shorter, and free from admixture interference observation polypide, effect is extremely protruded.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of dyeing of pyriform worm blood film compound stain solution and blood film
Method.
Background technology
Piroplasmosis(piroplasmosis), also known as " babesiasis " or " haemosoridiasis ".It is a class through propagating firmly
Hematophagia protozoosis, including Taylor worm(Theileria app)And Babesia(Babesia app)Caused blood is former
The general name of parasitosis.The domestic animals such as ox, horse, sheep and rodent and other wild animals are more common in, this disease is clinically delaied
Not in time, the death rate is high for the symptoms such as heat, anaemia, expiratory dyspnea, jaundice, such as diagnosis and treatment, and this disease is widely current all over the world, gives
Animal husbandry and national economy bring about great losses, according to FAO (Food and Agriculture Organization of the United Nation)(FAO)It is annual that statistics, tick and tick pass bloodprotozoonoses
About 7,000,000,000 dollars of economic loss can be caused to Animal husbandry production.The disease is widely distributed in China, and national 31 provinces and cities are almost
There is the report of this sick generation or pathogen separation, the production to China's animal husbandry causes huge economic loss.It is estimated that I
State there are about 35,000,000 every year(Only)Small ruminant infects piroplasmosis, and direct year economic loss is up to hundred million yuan of 3-4.Due to this
The cause of disease of disease is propagated by hard tick, so once incoming would become hard to eliminate.Mentioned at present in relevant document and use blood smear pin
Polypide is dyed, its colouring method is excessively complicated, and the used time is longer, and Color is undesirable, and staining impurity is more, shadow
Ring observation polypide.
The content of the invention
An object of the present invention:A kind of pyriform worm blood film compound stain solution formula is developed, using this with can be fine
Pyriform worm is dyed, is easy to observe and imparts knowledge to students;Two be exactly to pass through colouring method energy reasonable in design using above-mentioned dyestuff
Preferably the pyriform worm in blood is dyed.
Technical scheme one:A kind of pyriform worm blood film compound stain solution, the compound stain solution includes compound stain solution I
With compound dye night II;Described compound stain solution I is by Rui Shi dyestuffs 2g, Jim Sa pigment 1.7g, glycerine 50ml and methanol
1000ml is constituted, and specific preparation method is:Rui Shi dyestuffs 2g and Jim Sa pigment 1.7g is added in mortar, plus 50ml glycerine is ground
Addition 1000ml methanol is worn into after homogeneous paste, grinds and collects upper strata dye liquor and be stored in brown bottle, you can;The compound dye
Liquid II is made up of AMSP 2.3g, anhydrous potassium dihydrogenphosphate 0.3g and 1000ml distilled water, specific to prepare
Method is:Dissolve, prepare in the distilled water that AMSP 2.3g, anhydrous potassium dihydrogenphosphate 0.3g are added to 1000ml
Go out pH values for 6.5-6.8 phosphate buffers.
Technical scheme two, a kind of colouring method of pyriform worm blood film, the colouring method comprise the steps:
First, the drop of blood that 20 μ l carry pyriform worm is drawn with pipettor, drips to slide A one end, separately take the smooth load glass of a block edge
Piece B is placed on drop of blood center, blood is opened along slide B edges are even as push jack, after that 45 ° of slide B is put into slide A is empty
White local push jack backward, makes to leave one layer of thin smear film on slide A pieces;Room temperature is placed after push jack success, dries blood film, is applied
The thickness of blood film is should be noted during piece, red cell distribution is uniform, it is ensured that red blood cell should not be accumulated, while between can not having in blood film
Gap, it is standby;
2nd, the blood film prepared in step one is placed into room temperature 5 minutes, be put into after blood is air-dried completely in methanol to red thin
Born of the same parents are fixed, and are again placed in room temperature 3 minutes, treat that methanol volatilization is clean, standby;
3rd, the blood film prepared in step 2 is put into 1-1.5min in compound stain solution I;Then take out blood film be put into it is compound
1-2min in dye liquor II;Finally gently rinsed with clear water again, room temperature, which dries to dye, is made pyriform worm blood film.
The material being related in above-mentioned technical proposal is commercially available.
Beneficial effect:The composite dye and colouring method of the present invention is right in the case where being not required to by high-end instrument and equipment
Preferably, economical and practical, required time is shorter for pyriform worm Color, and free from admixture interference observation polypide, effect is extremely protruded.
Brief description of the drawings
Accompanying drawing 1 is this blood film of Niu Shuanya BABEIs;Accompanying drawing 2 is Niu Babeisi blood films;Accompanying drawing 3 is horse Babesia caballi blood
Smear.
Embodiment
Embodiment 1, a kind of pyriform worm blood film compound stain solution, the compound stain solution include compound stain solution I and compound dye night
Ⅱ;Described compound stain solution I is made up of Rui Shi dyestuffs 2g, Jim Sa pigment 1.7g, glycerine 50ml and methanol 1000ml, specifically
Preparation method is:Rui Shi dyestuffs 2g and Jim Sa pigment 1.7g is added in mortar, plus 50ml glycerine is ground to form after homogeneous paste
1000ml methanol is added, grinds and collects upper strata dye liquor and be stored in brown bottle, you can;The compound stain solution II is by anhydrous phosphorus
Acid dihydride sodium 2.3g, anhydrous potassium dihydrogenphosphate 0.3g and 1000ml distilled water are constituted, and specific preparation method is:By anhydrous phosphorus
Acid dihydride sodium 2.3g, anhydrous potassium dihydrogenphosphate 0.3g are added to be dissolved in 1000ml distilled water, prepares pH values for 6.5-6.8
Phosphate buffer.
Embodiment 2, a kind of colouring method of pyriform worm blood film, the colouring method comprise the steps:
First, the drop of blood that 20 μ l carry pyriform worm is drawn with pipettor, drips to slide A one end, separately take the smooth load glass of a block edge
Piece B is placed on drop of blood center, blood is opened along slide B edges are even as push jack, after that 45 ° of slide B is put into slide A is empty
White local push jack backward, makes to leave one layer of thin smear film on slide A pieces;Room temperature is placed after push jack success, dries blood film, is applied
The thickness of blood film is should be noted during piece, red cell distribution is uniform, it is ensured that red blood cell should not be accumulated, while between can not having in blood film
Gap, it is standby;
2nd, the blood film prepared in step one is placed into room temperature 5 minutes, be put into after blood is air-dried completely in methanol to red thin
Born of the same parents are fixed, and are again placed in room temperature 3 minutes, treat that methanol volatilization is clean, standby;
3rd, the blood film prepared in step 2 is put into 1-1.5min in compound stain solution I;Then take out blood film be put into it is compound
1-2min in dye liquor II;Finally gently rinsed with clear water again, room temperature, which dries to dye, is made pyriform worm blood film.
Embodiment 3, can be bright with the blood film prepared by above-mentioned technical method as shown in accompanying drawing 1, accompanying drawing 2 and accompanying drawing 3
Aobvious observes pyriform worm under the microscope.
Claims (2)
1. a kind of pyriform worm blood film compound stain solution, it is characterised in that:The compound stain solution includes compound stain solution I and compound dye night
Ⅱ;Described compound stain solution I is made up of Rui Shi dyestuffs 2g, Jim Sa pigment 1.7g, glycerine 50ml and methanol 1000ml, specifically
Preparation method is:Rui Shi dyestuffs 2g and Jim Sa pigment 1.7g is added in mortar, plus 50ml glycerine is ground to form after homogeneous paste
1000ml methanol is added, grinds and collects upper strata dye liquor and be stored in brown bottle, you can;The compound stain solution II is by anhydrous phosphorus
Acid dihydride sodium 2.3g, anhydrous potassium dihydrogenphosphate 0.3g and 1000ml distilled water are constituted, and specific preparation method is:By anhydrous phosphorus
Acid dihydride sodium 2.3g, anhydrous potassium dihydrogenphosphate 0.3g are added to be dissolved in 1000ml distilled water, prepares pH values for 6.5-6.8
Phosphate buffer, above-mentioned used dyestuff and reagent are conventional material.
2. a kind of colouring method of pyriform worm blood film, it is characterised in that:The colouring method comprises the steps:
First, the drop of blood that 20 μ l carry pyriform worm is drawn with pipettor, drips to slide A one end, separately take the smooth load glass of a block edge
Piece B is placed on drop of blood center, blood is opened along slide B edges are even as push jack, after that 45 ° of slide B is put into slide A is empty
White local push jack backward, makes to leave one layer of thin smear film on slide A pieces;Room temperature is placed after push jack success, dries blood film, is applied
The thickness of blood film is should be noted during piece, red cell distribution is uniform, it is ensured that red blood cell should not be accumulated, while between can not having in blood film
Gap, it is standby;
2nd, the blood film prepared in step one is placed into room temperature 5 minutes, be put into after blood is air-dried completely in methanol to red thin
Born of the same parents are fixed, and are again placed in room temperature 3 minutes, treat that methanol volatilization is clean, standby;
3rd, the blood film prepared in step 2 is put into 1-1.5min in compound stain solution I;Then take out blood film be put into it is compound
1-2min in dye liquor II;Finally gently rinsed with clear water again, room temperature, which dries to dye, is made pyriform worm blood film.
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CN201710450703.8A CN107300495A (en) | 2017-06-15 | 2017-06-15 | A kind of colouring method of pyriform worm blood film compound stain solution and blood film |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108414327A (en) * | 2017-10-31 | 2018-08-17 | 天津协和华美医学诊断技术有限公司 | A kind of Wright-Giemsa staining reagents and its application method |
CN108663252A (en) * | 2018-08-14 | 2018-10-16 | 苏州丰泰医疗用品贸易有限公司 | A kind of method of quick carry out Rui Shi Giemsa stainings |
CN109799123A (en) * | 2018-12-29 | 2019-05-24 | 广州和能生物科技有限公司 | A kind of method of quick carry out Rui Shi Giemsa staining |
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US3870146A (en) * | 1973-12-10 | 1975-03-11 | Sci Med Lab Inc | Wright{3 s stain packet |
CN1434127A (en) * | 2002-01-22 | 2003-08-06 | 裴芳君 | Staining kit and preparation method, staining method, and use thereof |
KR20030066520A (en) * | 2003-07-11 | 2003-08-09 | 서인범 | Cell Count Method For Low Cell Concentration |
WO2013101776A2 (en) * | 2011-12-28 | 2013-07-04 | Abbott Laboratories | Accelerated wright-giemsa and may-grünwald staining methods |
CN103323313A (en) * | 2012-03-23 | 2013-09-25 | 黄伏生 | Liquid-based cell analyzer staining method of cells exfoliated from serous cavity by using Wright staining method |
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2017
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KR20030066520A (en) * | 2003-07-11 | 2003-08-09 | 서인범 | Cell Count Method For Low Cell Concentration |
WO2013101776A2 (en) * | 2011-12-28 | 2013-07-04 | Abbott Laboratories | Accelerated wright-giemsa and may-grünwald staining methods |
CN103323313A (en) * | 2012-03-23 | 2013-09-25 | 黄伏生 | Liquid-based cell analyzer staining method of cells exfoliated from serous cavity by using Wright staining method |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108414327A (en) * | 2017-10-31 | 2018-08-17 | 天津协和华美医学诊断技术有限公司 | A kind of Wright-Giemsa staining reagents and its application method |
CN108663252A (en) * | 2018-08-14 | 2018-10-16 | 苏州丰泰医疗用品贸易有限公司 | A kind of method of quick carry out Rui Shi Giemsa stainings |
CN109799123A (en) * | 2018-12-29 | 2019-05-24 | 广州和能生物科技有限公司 | A kind of method of quick carry out Rui Shi Giemsa staining |
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Application publication date: 20171027 |