CN108663253A - One kind quickly carrying out Gram-stained method - Google Patents

One kind quickly carrying out Gram-stained method Download PDF

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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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CN
China
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
Application number
CN201810921434.3A
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Chinese (zh)
Inventor
殷跃锋
王云
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Suzhou Fengtai Medical Supplies Trading Co Ltd
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Suzhou Fengtai Medical Supplies Trading Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Suzhou Fengtai Medical Supplies Trading Co Ltd filed Critical Suzhou Fengtai Medical Supplies Trading Co Ltd
Priority to CN201810921434.3A priority Critical patent/CN108663253A/en
Publication of CN108663253A publication Critical patent/CN108663253A/en
Pending legal-status Critical Current

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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N2001/302Stain compositions

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  • 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

One kind quickly carrying out Gram-stained method
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.
CN201810921434.3A 2018-08-14 2018-08-14 One kind quickly carrying out Gram-stained method Pending CN108663253A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810921434.3A CN108663253A (en) 2018-08-14 2018-08-14 One kind quickly carrying out Gram-stained method

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810921434.3A CN108663253A (en) 2018-08-14 2018-08-14 One kind quickly carrying out Gram-stained method

Publications (1)

Publication Number Publication Date
CN108663253A true CN108663253A (en) 2018-10-16

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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
CN (1) CN108663253A (en)

Citations (10)

* Cited by examiner, † Cited by third party
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

Patent Citations (10)

* Cited by examiner, † Cited by third party
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)

* Cited by examiner, † Cited by third party
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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