Dyeing method for bacterial capsules in Haemophilus influenzae type b fermentation broth

A technology of Haemophilus influenzae and staining method, which is applied in the field of bacterial capsule staining of Haemophilus influenzae type b fermentation broth, which can solve the problems of high water content of bacterial capsule, high requirements for operation skills, and influence on observation results, etc. , to achieve the effect of good dyeing effect, low operating skill requirements and easy observation

Inactive Publication Date: 2012-10-17
CHENGDU OLYMVAX BIOPHARM
View PDF3 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, the operation process is relatively complicated, which requires high operating skills of the experimenter, which may lead to the failure of the experiment; the stability of the dye is also high, and if a large particle dye is used, the observation result will often be affected; in addition, these After the method is stained, often only a few bacteria can be seen, and it needs to be searched in a large area under the microscope, which is very inconvenient for actual operation
Moreover, due to the high water content of the bacterial capsule, heating will cause the bacteria to lose water and shrink, and will separate from the dye around the cells to produce a transparent bright area, causing some non-capsulated bacteria to be mistaken for capsuled bacteria.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Dyeing method for bacterial capsules in Haemophilus influenzae type b fermentation broth
  • Dyeing method for bacterial capsules in Haemophilus influenzae type b fermentation broth
  • Dyeing method for bacterial capsules in Haemophilus influenzae type b fermentation broth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A method for staining the bacterial capsule of the fermentation broth of type b Haemophilus influenzae includes the following steps:

[0034] A. Pretreatment: Take 1ml of the fermentation broth of Haemophilus influenzae type b and cool it in an ice bath for 0.5 min;

[0035] B. Preparation: Use a 0-20μL pipette to take 4μL of pretreated Haemophilus influenzae type b fermentation broth and drop it on the upper left corner of the slide, and drop 4μL of drawing ink on the lower left corner of the slide to make influenza b Haemophilus fermentation broth is mixed with drawing ink evenly;

[0036] C. Smear: Take a clean cover glass and make it form a 45° angle with the glass slide, then scrape the mixture from the left to the right;

[0037] D. Drying: Place the slides in the air to dry naturally;

[0038] E. Staining: After drying, drop 1 drop of 0.5% sahuang solution on the slide. After fixing for 1.5 min, wash off the smear with sahuang solution with water;

[0039] F. Seconda...

Embodiment 2

[0042] A method for staining the bacterial capsule of the fermentation broth of type b Haemophilus influenzae includes the following steps:

[0043] A. Pretreatment: Take 1ml of the fermentation broth of Haemophilus influenzae type b and cool it in an ice bath for 1 min;

[0044] B. Preparation: Use a 0-20μL pipette to take 5μL of the pretreated Haemophilus influenzae type b fermentation broth and drop it on the upper left corner of the slide, and drop 5μL of drawing ink onto the lower left corner of the slide to make influenza b Haemophilus fermentation broth is mixed with drawing ink evenly;

[0045] C. Smear: Take a clean cover glass and make it form a 45° angle with the glass slide, then scrape the mixture from the left to the right;

[0046] D. Drying: Place the slides in the air to dry naturally;

[0047] E. Staining: After drying, drop 2 drops of 0.5% sahuang solution on the slide, fix for 2 minutes and wash off the smear with water;

[0048] F. Secondary drying: absorb the...

Embodiment 3

[0051] A method for staining the bacterial capsule of the fermentation broth of type b Haemophilus influenzae includes the following steps:

[0052] A. Pretreatment: Take 1ml of the fermentation broth of Haemophilus influenzae type b and cool it in an ice bath for 1.5 minutes;

[0053] B. Preparation: Use a 0-20μL pipette to take 6μL of pretreated Haemophilus influenzae type b fermentation broth and drop it on the upper left corner of the slide, and drop 6μL of drawing ink on the lower left corner of the slide to make influenza b Haemophilus fermentation broth is mixed with drawing ink evenly;

[0054] C. Smear: Take a clean cover glass, and make it an angle of 45° with the glass slide, and then scrape the mixture from the left to the right;

[0055] D. Drying: Place the slides in the air to dry naturally;

[0056] E. Staining: After drying, drop 3 drops of 0.5% sahuang solution on the slide, fix for 2.5 minutes and then wash off the smear with water;

[0057] F. Secondary drying...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a dyeing method for bacterial capsules in Haemophilus influenzae type b fermentation broth. The method comprises the steps of pretreatment, smear preparation, smearing, drying, dyeing, secondary drying and microscopic examination The method employs the principle of negative dyeing, which guarantees that bacterial capsules are integral and thalluses occupy a great area and are easy to identify; a good dyeing effect is obtained, and color contrast between the capsules and a background is obvious, so the capsules can be easily observed; operation process is simple and does not have high requirements on operating skill of a tester, so the success rate of testing is high.

Description

Technical field [0001] The invention relates to a bacterial staining method, in particular to a bacterial capsule staining method of type b Haemophilus influenzae fermentation broth. Background technique [0002] Haemophilus influenzae (scientific name Haemophilus influenzae), referred to as Haemophilus influenzae, formerly known as Pfeifferella (or Baifeierella) or influenza bacillus, belongs to the genus Haemophilus of Vibrioceae, and is a kind of non-motile Gram-negative bacilli. [0003] Haemophilus influenzae is classified into two types: capsular strain and non-capsular strain. Among them, the capsular Haemophilus influenzae type b, namely Haemophilus influenzae type b (referred to as Hib) is one of the main causes of toxicity. In order to prevent invasive infections caused by Haemophilus influenzae, a type b Haemophilus influenzae conjugate vaccine made by covalently binding purified Hib capsular polysaccharide and tetanus toxoid protein is often used to prevent it. [0004...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N1/30
Inventor 朱琳韩炼赵丹罗芹赵涛
Owner CHENGDU OLYMVAX BIOPHARM
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap