CN108663253A - One kind quickly carrying out Gram-stained method - Google Patents
One kind quickly carrying out Gram-stained method Download PDFInfo
- Publication number
- CN108663253A CN108663253A CN201810921434.3A CN201810921434A CN108663253A CN 108663253 A CN108663253 A CN 108663253A CN 201810921434 A CN201810921434 A CN 201810921434A CN 108663253 A CN108663253 A CN 108663253A
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- Prior art keywords
- gram
- liquid
- distilled water
- alcohol
- iodine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 16
- 239000007788 liquid Substances 0.000 claims abstract description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000012153 distilled water Substances 0.000 claims abstract description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 21
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 claims abstract description 21
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229910052740 iodine Inorganic materials 0.000 claims abstract description 13
- 239000011630 iodine Substances 0.000 claims abstract description 13
- 235000019441 ethanol Nutrition 0.000 claims abstract description 12
- JUQPZRLQQYSMEQ-UHFFFAOYSA-N CI Basic red 9 Chemical compound [Cl-].C1=CC(N)=CC=C1C(C=1C=CC(N)=CC=1)=C1C=CC(=[NH2+])C=C1 JUQPZRLQQYSMEQ-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229940052223 basic fuchsin Drugs 0.000 claims abstract description 7
- 239000013078 crystal Substances 0.000 claims abstract description 6
- VBIXEXWLHSRNKB-UHFFFAOYSA-N ammonium oxalate Chemical compound [NH4+].[NH4+].[O-]C(=O)C([O-])=O VBIXEXWLHSRNKB-UHFFFAOYSA-N 0.000 claims abstract description 4
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 abstract description 11
- 238000004043 dyeing Methods 0.000 abstract description 7
- 238000010186 staining Methods 0.000 abstract description 4
- 241000192125 Firmicutes Species 0.000 abstract description 2
- 238000005406 washing Methods 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 1
- 241000233855 Orchidaceae Species 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004061 bleaching Methods 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000004042 decolorization Methods 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses one kind of Gram's staining technical field quickly to carry out Gram-stained method;Gram A liquid:A liquid:Crystal violet 1g, 95% alcohol 20ml, B liquid:Ammonium oxalate 0.8g distilled water 80ml;Gram B liquid:Crystalline flake of iodine 1g, potassium iodide 2g, distilled water 300ml, gram C liquid:Absolute ethyl alcohol 95ml, distilled water 5m, gram D liquid:Husky of common dye 0.25g, basic fuchsin 0.1g, 95% alcohol 10ml, distilled water 90ml;This method dyeing time is short, and clinic can be diagnosed quickly, differentiates gram-positive bacteria and negative bacterium.
Description
Technical field
The present invention relates to Gram's stain technical fields, specially a kind of quickly to carry out Gram-stained method.
Background technology
Gram's stain refers to widely used a kind of important differential staining in bacteriology, belongs to staining counter.This
Kind decoration method was invented in 1884 by Danish doctor leather orchid, and Gram staining generally comprises first dye, mordant dyeing, decolourizes, redyes
Deng four steps.The bacterium of no dyeing, since it is very small with ambient enviroment index of refraction difference, therefore extremely difficult area under the microscope
Not.After dyed, positive bacteria is in purple, and negative bacterium takes on a red color, and can clearly be observed that form, arrangement and the certain knots of bacterium
Structure feature, to taxonomic identification.Dyeing theory:After dye at the beginning of crystal violet and iodine solution mordant dyeing, the shape in bacteria cell wall
At the compound of crystal violet and iodine not soluble in water, then with 95% ethanol decolorization, but when traditional Gram's stain dyeing
Between it is long, the completion of specimen staining needs the about 5min times, and decoloration is more demanding to time control, and bleaching time is insufficient, cloudy
Property bacterium accidentally dyed positive bacteria, decoloration is excessive, and positive bacteria is accidentally dyed negative bacterium, for this purpose, it is proposed that a kind of quickly removed from office
The method of blue Albert'stain Albert.
Invention content
The purpose of the present invention is to provide one kind quickly carrying out Gram-stained method, to solve in above-mentioned background technology
The problem of proposition.
To achieve the above object, the present invention provides the following technical solutions:One kind quickly carrying out Gram-stained method:
Gram A liquid:
A liquid:Crystal violet 1g,
95% alcohol 20ml,
B liquid:Ammonium oxalate 0.8g,
Distilled water 80ml;
Gram B liquid:
Crystalline flake of iodine 1g,
Potassium iodide 2g,
Distilled water 300ml;
Gram C liquid:
Absolute ethyl alcohol 95ml,
Distilled water 5m;
Gram D liquid:
Husky of common dye 0.25g,
Basic fuchsin 0.1g,
95% alcohol 10ml,
Distilled water 90ml.
Compared with prior art, the beneficial effects of the invention are as follows:This method dyeing time is short, and clinic can be examined quickly
It is disconnected, differentiate gram-positive bacteria and negative bacterium.
Specific implementation mode
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described,
Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, every other embodiment obtained by those of ordinary skill in the art without making creative efforts, all
Belong to the scope of protection of the invention.
Embodiment:
Gram A liquid:
A:A liquid:Crystal violet 1g, 95% alcohol 20ml;B liquid:Ammonium oxalate 0.8g, distilled water 80ml;
B:Bis- liquid of A, B is mixed, standing uses afterwards for 24 hours;
Gram B liquid:
C:Potassium iodide is first dissolved in a small amount of water, then the crystalline flake of iodine is dissolved in by crystalline flake of iodine 1g, potassium iodide 2g, distilled water 300ml
In liquor kalii iodide, after iodine is entirely molten, moisture is filled up;
Gram C liquid:
D:Absolute ethyl alcohol 95ml is taken, 5ml distilled water, mixing is added;
Gram D liquid:
E:Husky of common dye 0.25g, basic fuchsin 0.1g, 95% alcohol 10ml, distilled water 90ml, husky of common dye and basic fuchsin are dissolved in
In ethyl alcohol, then diluted with distilled water.
Wherein, the gram B liquid needs that first potassium iodide 2g is dissolved in a small amount of distilled water, then crystalline flake of iodine 1g is dissolved in iodine
Change in potassium solution, after iodine is entirely molten, suitable distilled water can just be added;The gram C liquid need to be by absolute ethyl alcohol 95ml and 5ml
Distilled water, which pours into, is sufficiently stirred mixing in container;The gram D liquid needs first to be dissolved in husky of common dye 0.25g and basic fuchsin 0.1g
95% alcohol, then diluted with distilled water is poured slowly into;It is described that sample is spread evenly across on clean slide, it spontaneously dries, flame
It is fixed;And this quickly carries out Gram-stained method and includes the following steps:
S1:Sample is spread evenly across on clean slide, is spontaneously dried, flame is fixed;
S2:Gram A liquid contaminates 10s, washing;
S3:Gram B liquid is added to cover coating surface about 10s, washing sucks moisture with blotting paper;
S4:Gram C liquid decoloration 20s, washing is added to suck moisture;
S5:Add gram D liquid to cover coating surface, dye 10s, washes, dry, microscopy.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding without departing from the principles and spirit of the present invention can carry out these embodiments a variety of variations, modification, replace
And modification, the scope of the present invention is defined by the appended.
Claims (5)
1. one kind quickly carrying out Gram-stained method, it is characterised in that:
Gram A liquid:
A liquid:Crystal violet 1g,
95% alcohol 20ml,
B liquid:Ammonium oxalate 0.8g,
Distilled water 80ml;
Gram B liquid:
Crystalline flake of iodine 1g,
Potassium iodide 2g,
Distilled water 300ml;
Gram C liquid:
Absolute ethyl alcohol 95ml,
Distilled water 5m;
Gram D liquid:
Husky of common dye 0.25g,
Basic fuchsin 0.1g,
95% alcohol 10ml,
Distilled water 90ml.
2. one kind according to claim 1 quickly carries out Gram-stained method, it is characterised in that:The gram B
Liquid needs that first potassium iodide 2g is dissolved in a small amount of distilled water, then crystalline flake of iodine 1g is dissolved in liquor kalii iodide, after iodine is entirely molten,
Suitable distilled water can be added.
3. one kind according to claim 1 quickly carries out Gram-stained method, it is characterised in that:The gram C
Absolute ethyl alcohol 95ml and 5ml distilled water need to be poured into and be sufficiently stirred mixing in container by liquid.
4. one kind according to claim 1 quickly carries out Gram-stained method, it is characterised in that:The gram D
Liquid needs that husky of common dye 0.25g and basic fuchsin 0.1g are first dissolved in 95% alcohol, then is diluted with distilled water is poured slowly into.
5. one kind according to claim 1 quickly carries out Gram-stained method, it is characterised in that:It is described that sample is equal
It is even to be coated on clean slide, it spontaneously dries, flame is fixed.
Priority Applications (1)
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CN201810921434.3A CN108663253A (en) | 2018-08-14 | 2018-08-14 | One kind quickly carrying out Gram-stained method |
Applications Claiming Priority (1)
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CN201810921434.3A CN108663253A (en) | 2018-08-14 | 2018-08-14 | One kind quickly carrying out Gram-stained method |
Publications (1)
Publication Number | Publication Date |
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CN108663253A true CN108663253A (en) | 2018-10-16 |
Family
ID=63789711
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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CN201810921434.3A Pending CN108663253A (en) | 2018-08-14 | 2018-08-14 | One kind quickly carrying out Gram-stained method |
Country Status (1)
Country | Link |
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CN (1) | CN108663253A (en) |
Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5827680A (en) * | 1994-10-11 | 1998-10-27 | Difco Laboratories | Three reagent gram staining method and kit |
CN1392266A (en) * | 2001-06-19 | 2003-01-22 | 张华� | Quick gram staining liquid and method |
CN101126689A (en) * | 2007-08-07 | 2008-02-20 | 叶鹰 | Gram stain two-step method |
CN102080121A (en) * | 2009-11-26 | 2011-06-01 | 赫莫格莱姆工业通讯产品医学产品有限公司 | Staining process and apparatus used in bacteriology |
CN102220236A (en) * | 2010-04-15 | 2011-10-19 | 叶元康 | Automatic staining instrument with Gram staining two-step method |
CN103674661A (en) * | 2012-09-12 | 2014-03-26 | 无锡飞伊生物科技有限公司 | Washing-free Gram staining method |
CN103674660A (en) * | 2012-09-12 | 2014-03-26 | 无锡飞伊生物科技有限公司 | Gram staining two-step method with one-time washing |
CN107064286A (en) * | 2017-01-16 | 2017-08-18 | 常州市疾病预防控制中心 | The method of Gram-positive and negative bacterium Mass Spectrometer Method kit and Rapid identification |
CN108181155A (en) * | 2012-03-19 | 2018-06-19 | 文塔纳医疗系统公司 | Improve the Gram-staining process of the decoloration to crystal violet-Surgidine of gramnegative bacterium |
CN108384274A (en) * | 2018-02-28 | 2018-08-10 | 邱永兴 | A kind of novel low fat dyestuff |
-
2018
- 2018-08-14 CN CN201810921434.3A patent/CN108663253A/en active Pending
Patent Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5827680A (en) * | 1994-10-11 | 1998-10-27 | Difco Laboratories | Three reagent gram staining method and kit |
CN1392266A (en) * | 2001-06-19 | 2003-01-22 | 张华� | Quick gram staining liquid and method |
CN101126689A (en) * | 2007-08-07 | 2008-02-20 | 叶鹰 | Gram stain two-step method |
CN102080121A (en) * | 2009-11-26 | 2011-06-01 | 赫莫格莱姆工业通讯产品医学产品有限公司 | Staining process and apparatus used in bacteriology |
CN102220236A (en) * | 2010-04-15 | 2011-10-19 | 叶元康 | Automatic staining instrument with Gram staining two-step method |
CN108181155A (en) * | 2012-03-19 | 2018-06-19 | 文塔纳医疗系统公司 | Improve the Gram-staining process of the decoloration to crystal violet-Surgidine of gramnegative bacterium |
CN103674661A (en) * | 2012-09-12 | 2014-03-26 | 无锡飞伊生物科技有限公司 | Washing-free Gram staining method |
CN103674660A (en) * | 2012-09-12 | 2014-03-26 | 无锡飞伊生物科技有限公司 | Gram staining two-step method with one-time washing |
CN107064286A (en) * | 2017-01-16 | 2017-08-18 | 常州市疾病预防控制中心 | The method of Gram-positive and negative bacterium Mass Spectrometer Method kit and Rapid identification |
CN108384274A (en) * | 2018-02-28 | 2018-08-10 | 邱永兴 | A kind of novel low fat dyestuff |
Non-Patent Citations (2)
Title |
---|
庞爱梅: "《检验技术与临床诊断》", 31 May 2017, 吉林科学技术出版社 * |
温洪宇: "《环境微生物学实验教程》", 31 August 2017, 中国矿业大学出版社 * |
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Application publication date: 20181016 |
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