Capsule staining of bacteria

A dyeing method and technology for bacterial capsules, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as experimental failure, high requirements for operating skills, and incompetence for novices, to reduce the difficulty of operation and achieve high efficiency. Dyeing method, the effect of maintaining integrity

Inactive Publication Date: 2008-09-03
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, the operation skills of the experimenter are very high, which may easily lead to the failure of the experiment.
The background of the black ink method is very dark. To make a single cell transparently displayed, it needs to be scraped to a micron-level thickness. The effective area of ​​film production is very small, and it is difficult for novices to do it
The second is that only a few bacteria can be seen in specimens made by traditional negative staining methods such as the black ink method, and it is necessar...

Method used

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  • Capsule staining of bacteria
  • Capsule staining of bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] 1) Put a small drop of sterile water on the slide;

[0060] 2) Tablet preparation: use an inoculation loop to take a small amount of bacteria that has been cultured for 24 hours, and spread slowly from the center to the outside in a spiral shape in a water-wet state to keep the capsule intact;

[0061] 3) Natural air drying;

[0062] 4) Add a drop of pure methanol dropwise, fix for 1 minute, tilt and pour off the methanol;

[0063] 5) Add a few drops of phenol fuchsin and stain for 1 minute;

[0064] 6) Cover glass;

[0065]7) Carefully absorb the excess liquid around the coverslip with absorbent paper, tilt the slide and absorb part of the staining solution under the coverslip to increase the adhesion of the coverslip and the light transmittance of the sample during microscopic examination;

[0066] 8) Oil immersion microscope inspection.

[0067] The observation results are attached figure 2 As shown in (a), the background is red and the capsule is colorless.

Embodiment 2

[0069] 1) Put a small drop of sterile water on the slide;

[0070] 2) Tablet preparation: use an inoculation loop to take a small amount of bacteria that has been cultured for 48 hours, and spread slowly from the center to the outside in a spiral shape in a wet state to keep the capsule intact;

[0071] 3) Natural air drying;

[0072] 4) Fix with pure methanol for 2 minutes, tilt and pour off the methanol;

[0073] 5) Add a few drops of phenol fuchsin and stain for 3 minutes;

[0074] 6) Cover glass;

[0075] 7) Carefully absorb the excess liquid around the coverslip with absorbent paper, tilt the slide and absorb part of the staining solution under the coverslip to increase the adhesion of the coverslip and the light transmittance of the sample during microscopic examination;

[0076] 8) Oil immersion microscope inspection. The observation results are attached figure 2 As shown in (b), the background is red and the capsule is colorless.

Embodiment 3

[0078] 1) Put a small drop of sterile water on the slide;

[0079] 2) Tablet preparation: use an inoculation loop to take a small amount of bacteria that has been cultured for 36 hours, and spread slowly from the center to the outside in a spiral shape in a wet state to keep the capsule intact;

[0080] 3) Natural air drying;

[0081] 4) Add 2 drops of pure methanol dropwise, fix for 1 minute, tilt and pour off the methanol;

[0082] 5) Add a few drops of phenol fuchsin for 2 minutes

[0083] 6) Cover glass;

[0084] 7) Carefully absorb the excess liquid around the coverslip with absorbent paper, tilt the slide and absorb part of the staining solution under the coverslip to increase the adhesion of the coverslip and the light transmittance of the sample during microscopic examination;

[0085] 8) Oil immersion microscope inspection. The observation results are attached figure 2 As shown in (c), the background is red and the capsule is colorless.

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Abstract

The invention relates to a bacterial capsule staining method, comprising the steps: 1) dropping a droplet of sterile water on a glass slide; 2) bacteria culturing for 24 to 48 hours, selecting the bacteria with a transfering loop, and slowly coating the glass slide in water moisture state spirally from the center to the outside to keep the capsule complete; 3) natural drying or blow drying with electric wind; 4) dropping 2-3 droplets of pure methanol, fixing for 1-2 minutes, and inclining to remove the methanol; 5) dropping carbolic acid fuchsin to stain for 1-3 minutes, and covering a cover glass; and 6) absorbing the liquid surrounding the cover glass with a piece of absorbent paper, inclining the slide glass to absorb partial staining liquid under the cover glass. By improvement on the slide making method, the staining method and the operation steps, the completeness of bacterial capsule is effectively kept, the growing state of all the bacterial capsule in the specimen can be clearly displayed, the operation difficulty of capsule staining is obviously reduced, and the reliability of bacterial capsule recognition is improved.

Description

technical field [0001] The invention relates to a bacterial capsule staining method, which adopts a unique staining method to stain the bacterial capsule, and aims at preparing samples for bacterial capsule morphology observation in microbial experiments and medical testing. Background technique [0002] The capsule is a layer of mucus or gelatinous substance that surrounds the bacterial cell, part of which is synthesized in the cytoplasm, and the main component is polysaccharides. The molecular composition and configuration of these polysaccharides are diverse, making its structure extremely complex and becoming a serum credit type basis. For example, Streptococcus pneumoniae can be divided into at least 85 serotypes according to the antigenicity of its capsular polysaccharide. A few strains are polypeptides, such as Bacillus anthracis and Yersinia pestis. The main functions of the capsule are: [0003] ① Anti-phagocytosis: Capsule can effectively resist the phagocytosis...

Claims

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Application Information

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IPC IPC(8): C12Q1/04
Inventor 周志军许陈云周小英
Owner ZHEJIANG UNIV
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