CN110501336A - The capsular swelling colouring method of streptococcus pneumonia - Google Patents
The capsular swelling colouring method of streptococcus pneumonia Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 43
- 238000004040 coloring Methods 0.000 title claims abstract description 29
- 201000005010 Streptococcus pneumonia Diseases 0.000 title claims description 52
- 241000193998 Streptococcus pneumoniae Species 0.000 title claims description 52
- 230000008961 swelling Effects 0.000 title claims description 41
- 239000000975 dye Substances 0.000 claims abstract description 52
- 238000010186 staining Methods 0.000 claims abstract description 24
- 239000012528 membrane Substances 0.000 claims description 34
- 239000000976 ink Substances 0.000 claims description 32
- 239000011521 glass Substances 0.000 claims description 25
- 239000007788 liquid Substances 0.000 claims description 20
- 238000000386 microscopy Methods 0.000 claims description 20
- 239000013078 crystal Substances 0.000 claims description 17
- 244000061458 Solanum melongena Species 0.000 claims description 16
- 241000894006 Bacteria Species 0.000 claims description 14
- VBIXEXWLHSRNKB-UHFFFAOYSA-N ammonium oxalate Chemical compound [NH4+].[NH4+].[O-]C(=O)C([O-])=O VBIXEXWLHSRNKB-UHFFFAOYSA-N 0.000 claims description 13
- 239000008354 sodium chloride injection Substances 0.000 claims description 13
- 239000013642 negative control Substances 0.000 claims description 12
- 239000000243 solution Substances 0.000 claims description 11
- RBTBFTRPCNLSDE-UHFFFAOYSA-N 3,7-bis(dimethylamino)phenothiazin-5-ium Chemical compound C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 RBTBFTRPCNLSDE-UHFFFAOYSA-N 0.000 claims description 9
- 229960000907 methylthioninium chloride Drugs 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 8
- 239000001045 blue dye Substances 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims 1
- 238000003756 stirring Methods 0.000 claims 1
- 239000002775 capsule Substances 0.000 abstract description 8
- 238000004043 dyeing Methods 0.000 abstract description 5
- 230000001580 bacterial effect Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 abstract description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 210000002966 serum Anatomy 0.000 description 8
- 235000019441 ethanol Nutrition 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 206010053615 Thermal burn Diseases 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 2
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 239000012047 saturated solution Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000606768 Haemophilus influenzae Species 0.000 description 1
- 244000178870 Lavandula angustifolia Species 0.000 description 1
- 235000010663 Lavandula angustifolia Nutrition 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 244000297179 Syringa vulgaris Species 0.000 description 1
- 235000004338 Syringa vulgaris Nutrition 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 229940047650 haemophilus influenzae Drugs 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000001102 lavandula vera Substances 0.000 description 1
- 235000018219 lavender Nutrition 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
Classifications
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- 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
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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- Chemical & Material Sciences (AREA)
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- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
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- Engineering & Computer Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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Abstract
The present invention provides a kind of simple, effective pneumococcal capsule colouring methods.Improve numerous disadvantages of existing capsule stain.It is characterized in that: improvement after colouring method have the characteristics that it is easy to operate, saving dyestuff, be easy to observe, dyeing effect more better than traditional capsule staining can be obtained, improve the quality of Bacterial stain, easy operation sequence, it is applicable not only to Experiment on Microbiology teaching, it is also convenient for batch making teaching specimen, is worth in teaching, scientific research, popularization clinically.
Description
Technical field
The invention belongs to field of biology, are related to a kind of capsular swelling colouring method of streptococcus pneumonia.
Background technique
Capsular swelling test be specifically bound using the capsular antigen of specific antisera and corresponding bacterium to be formed it is compound
Object, to detect streptococcus pneumonia, haemophilus influenzae etc..
Current capsule stain and capsular swelling colouring method include:
1, Hiss anthony'S capsule stain Anthony method
Dye liquor: a, violet staining liquid: crystal violet ethyl alcohol saturated solution (2g crystal violet is dissolved in 95% ethyl alcohol of 20ml) 5ml,
Add water 95ml；B, 20% copper sulphate dye liquor
Method: a, nacterial smear, air spontaneously dry, and ethyl alcohol is fixed；B, Hiss dyeing liquor is added dropwise on slide, on flame
It until heating makes to have steam to emerge or dyes, does not wash；C, it is rinsed with 20% copper-bath, is not washed；D, naturally dry
Or blotting paper blots, in oily microscopic observation pod membrane
As a result: pod membrane is lavender or blue, and thallus is purple.
2, Hiss anthony'S capsule stain Anthony method is improved
Dye liquor: a, violet staining liquid: crystal violet ethyl alcohol saturated solution (2g crystal violet is dissolved in 95% ethyl alcohol of 20ml) 5ml,
Add water 95ml；B, 20% copper sulphate dye liquor.
Method: a, nacterial smear, air spontaneously dry, and methanol is fixed；B be added dropwise Hiss dyeing liquor in slide paint 2min,
Do not wash；C is rinsed with 20% copper-bath, is not washed；D naturally dry or blotting paper blot, in oily microscopic observation pod membrane.
As a result: pod membrane is in light red, and thallus is in deep blue purple color, and background is uniform lilac.
3, capsular swelling dyes
Dye liquor: 1% methylene blue solution
Method: a, taking 2 glass slides, and 5 μ l of culture of streptococcus pneumonia liquid is added dropwise respectively；B, corresponding pneumonia blood is added dropwise respectively
Cleer and peaceful 0.85% sodium chloride injection is mixed with bacterium solution；C, 5 μ l, 1% methylene blue staining liquid is added dropwise to mixed liquor respectively and mixes
It is even, coverslip is covered, 5-10 minutes postposition microscope oil is stored at room temperature in wet box under the microscope.
As a result: visible distinct, colourless endless belt, the i.e. pod membrane of swelling around thallus are judged to the positive；Nothing around thallus
It can be seen that the colourless endless belt of distinct, as capsular swelling are negative.
But above method is unable to get accurate detection in the actual operation process as a result, the probability of success is lower, often
It will appear thallus to be aggregated with serum, agglomerate；Thallus is floated in liquid and can not be observed；The too shallow nothing of dye liquor color around bacterium
Method sees phenomena such as pod membrane, and the accuracy rate and stability of detection thus greatly reduces.
Summary of the invention
The purpose of the present invention is to provide a kind of capsular swelling colouring methods of streptococcus pneumonia, can be obtained by this method
The testing result high to accuracy, stability is good.
Capsular swelling colouring method of the present invention, comprising the following steps:
1) 2 glass slides are taken, culture of streptococcus pneumonia liquid is added dropwise respectively；
2) corresponding streptococcus pneumonia specific antisera is added dropwise respectively and sodium chloride injection is mixed with bacterium solution, makes respectively
It is sufficiently stirred evenly with micropipettor pipette tips, and drop is tiled, be placed at room temperature for reaction, naturally dry is fixed through flame；
3) it dyes, ammonium oxalate crystal violet dye liquor or methylene blue dye liquor, washing, blotting paper suck dry moisture is added dropwise；
4) negative staining draws a drop prepared Chinese ink and is added drop-wise to glass slide side, tilts specimen slide, contacted with another piece of glass slide
Prepared Chinese ink and with specimen slide at 45 degree of angles, oliquely downward drawing gently, so that prepared Chinese ink covering coloring face, last room temperature is naturally dry
It is dry；
5) microscopy, oil mirror observation.
Capsular swelling colouring method of the present invention, further includes further deterministic process:
Streptococcus pneumonia thallus dyes aubergine or blue, has a colorless and transparent pod membrane area outside thallus, background is black, bacterium
Body, background, pod membrane comparison clearly, are judged to the positive；
Streptococcus pneumonia thallus dyes aubergine or blue, can not see transparent pod membrane area outside thallus, black background with
Thallus is connected, as capsular swelling negative control.
Wherein, the concentration of sodium chloride injection is 0.9%；
Wherein, the concentration of prepared Chinese ink is 10%.
Preferably, colouring method of the present invention, which comprises the following steps:
1) 2 glass slides are taken, 5 μ l of culture of streptococcus pneumonia liquid is added dropwise respectively；
2) 5 μ l of streptococcus pneumonia specific antisera and sodium chloride corresponding to different streptococcus pneumonia types are added dropwise respectively
5 μ l of injection is mixed with bacterium solution, is sufficiently stirred evenly using micropipettor pipette tips, and drop is tiled respectively, is placed at room temperature for reaction,
Naturally dry is fixed through flame；
3) it dyes, ammonium oxalate crystal violet dye liquor is added dropwise or methylenum careuleum dye liquor dyes 1 minute, washing, blotting paper suck dry moisture；
4) negative staining draws a drop prepared Chinese ink and is added drop-wise to glass slide side, tilts specimen slide, contacted with another piece of glass slide
Prepared Chinese ink and with specimen slide at 45 degree of angles, oliquely downward drawing gently, so that prepared Chinese ink covering coloring face, last room temperature is naturally dry
It is dry；
5) microscopy, oil mirror observation；
Capsular swelling colouring method of the present invention, further includes further deterministic process:
Streptococcus pneumonia thallus dyes aubergine or blue, has a colorless and transparent pod membrane area outside thallus, background is black, bacterium
Body, background, pod membrane comparison clearly, are judged to the positive；
Streptococcus pneumonia thallus dyes aubergine or blue, can not see transparent pod membrane area outside thallus, black background with
Thallus is connected, as capsular swelling negative control.
Method of the invention is improved in the following areas:
1, serum is reacted with bacterium solution, i.e., specific antisera is formed compound in conjunction with S. pneumoniae capsular antigentic specificity
Object, and make pod membrane that swelling occur, convenient for observation pod membrane.And the sodium chloride injection of negative control will not occur with streptococcus pneumonia
Capsule swelling reaction is difficult to observe that S. pneumoniae capsular whether there is.
Naturally dry and fixation, are conducive to next dyeing and microscopy step in this way, and existing capsular swelling test,
Thallus can not be fixed in liquid, when microscopy the more difficult thallus for capturing capsular swelling.
2, it the use of the purpose of ammonium oxalate crystal violet dye liquor or methylene blue dye liquor is dyed to thallus, is seen convenient for microscopy
It examines.
3, the use of the purpose of prepared Chinese ink negative staining dyed to background, can be observed in microscopy under black background with purple
Transparent pod membrane between color or the thallus of blue achievees the purpose that observe pod membrane.
After the present invention by improveing above, a kind of capsular swelling colouring method of completely new streptococcus pneumonia is provided.The party
Method have the characteristics that it is easy to operate, save dyestuff, be easy to observe, can obtain and preferably dye effect than traditional capsule staining
Fruit improves the quality of Bacterial stain, and easy operation sequence is applicable not only to Experiment on Microbiology teaching, is also convenient for batch and makes
Make teaching specimen, is worth in teaching, scientific research, popularization clinically.
Detailed description of the invention
Fig. 1 is prepared Chinese ink negative staining process
Fig. 2 is the photo of 15B type streptococcus pneumonia gram stain microscopy.
Fig. 3 is the microscopy photo of the 15B type S. pneumoniae capsular swelling negative staining positive.
Fig. 4 is the microscopy photo of 15B type S. pneumoniae capsular swelling negative staining negative control.
Fig. 5 is the photo of 17F type streptococcus pneumonia gram stain microscopy.
Fig. 6 is the microscopy photo of the 17F type S. pneumoniae capsular swelling negative staining positive.
Fig. 7 is the microscopy photo of 17F type S. pneumoniae capsular swelling negative staining negative control.
Fig. 8 is the microscopy photo of the 19F type S. pneumoniae capsular swelling negative staining positive.
Fig. 9 is the microscopy photo of 19F type S. pneumoniae capsular swelling negative staining negative control.
Figure 10 is existing capsular swelling dyeing microscopic examination figure.
Specific embodiment
By following specific embodiments, the present invention is further illustrated, but uses not as present invention limitation.
Embodiment 1, capsular swelling colouring method
1, material
Culture of streptococcus pneumonia liquid, specific antisera, ammonium oxalate crystal violet dye liquor, methylene blue dye liquor, sodium chloride note
Penetrate liquid, 10% prepared Chinese ink
2, method
1) 2 glass slides are taken, 5 μ l of culture of streptococcus pneumonia liquid is added dropwise respectively；
2) corresponding 5 μ l of streptococcus pneumonia specific antisera is added dropwise respectively and 5 μ l of sodium chloride injection is mixed with bacterium solution,
It is sufficiently stirred evenly using micropipettor pipette tips respectively, and drop is tiled, be placed at room temperature for reaction, naturally dry is fixed through flame
(non-scald on hand is advisable)；
3) it dyes, ammonium oxalate crystal violet dye liquor is added dropwise or methylenum careuleum dye liquor dyes 1 minute, washing, blotting paper suck dry moisture；
4) negative staining draws a drop prepared Chinese ink and is added drop-wise to glass slide side, tilts specimen slide, contacted with another piece of glass slide
Prepared Chinese ink and with specimen slide at 45 degree of angles, oliquely downward drawing gently, so that prepared Chinese ink covering coloring face, last room temperature is naturally dry
It is dry.
5) microscopy, oil mirror observation.
3, result
Streptococcus pneumonia thallus is dyed aubergine (or blue), has a colorless and transparent pod membrane area outside thallus, and background is black,
Thallus, background, pod membrane comparison clearly, are judged to the positive；
Streptococcus pneumonia thallus is dyed aubergine (or blue), can not see transparent pod membrane area, black background outside thallus
It is connected with thallus, as capsular swelling negative control.
Embodiment 2,15B type S. pneumoniae capsular swelling method
1, material
15B type culture of streptococcus pneumonia liquid, 15B type streptococcus pneumonia antiserum (factor serum 15h), ammonium oxalate
Crystal violet dye liquor, methylene blue dye liquor, sodium chloride injection, 10% prepared Chinese ink
2, method
1) 2 glass slides are taken, 5 μ l of 15B type culture of streptococcus pneumonia liquid is added dropwise respectively；
2) 15B type streptococcus pneumonia antiserum (factor serum 15h) 5 μ l and 5 μ l of sodium chloride injection are added dropwise respectively
It mixes, is sufficiently stirred evenly using micropipettor pipette tips respectively, and drop is tiled with bacterium solution, be placed at room temperature for reaction, naturally dry,
(non-scald on hand is advisable) is fixed through flame；
3) it dyes, ammonium oxalate crystal violet dye liquor is added dropwise or methylenum careuleum dye liquor dyes 1 minute, washing, blotting paper suck dry moisture；
4) negative staining draws a drop prepared Chinese ink and is added drop-wise to glass slide side, tilts specimen slide, contacted with another piece of glass slide
Prepared Chinese ink and with specimen slide at 45 degree of angles, oliquely downward drawing gently, so that prepared Chinese ink covering coloring face, last room temperature is naturally dry
It is dry.
5) microscopy, oil mirror observation.
3, result
15B type streptococcus pneumonia thallus is dyed aubergine (or blue), has a colorless and transparent pod membrane area outside thallus, background is
Black, thallus, background, pod membrane comparison clearly, are judged to the positive；
15B type streptococcus pneumonia thallus is dyed aubergine (or blue), can not see transparent pod membrane area, black outside thallus
Background is connected with thallus, as capsular swelling negative control.
Fig. 2-4 is the 15B type S. pneumoniae capsular swelling negative staining positive and negative photo.
Embodiment 3,17F type S. pneumoniae capsular swelling colouring method
1, material
17F type culture of streptococcus pneumonia liquid, 17F type streptococcus pneumonia antiserum (factor serum 17b), ammonium oxalate
Crystal violet dye liquor, methylene blue dye liquor, sodium chloride injection, 10% prepared Chinese ink
2, method
1) 2 glass slides are taken, 5 μ l of 17F type culture of streptococcus pneumonia liquid is added dropwise respectively；
2) 17F type streptococcus pneumonia antiserum (factor serum 15h) 5 μ l and 5 μ l of sodium chloride injection are added dropwise respectively
It mixes, is sufficiently stirred evenly using micropipettor pipette tips respectively, and drop is tiled with bacterium solution, be placed at room temperature for reaction, naturally dry,
(non-scald on hand is advisable) is fixed through flame；
3) it dyes, ammonium oxalate crystal violet dye liquor is added dropwise or methylenum careuleum dye liquor dyes 1 minute, washing, blotting paper suck dry moisture；
4) negative staining draws a drop prepared Chinese ink and is added drop-wise to glass slide side, tilts specimen slide, contacted with another piece of glass slide
Prepared Chinese ink and with specimen slide at 45 degree of angles, oliquely downward drawing gently, so that prepared Chinese ink covering coloring face, last room temperature is naturally dry
It is dry.
5) microscopy, oil mirror observation.
3, result
17F type streptococcus pneumonia thallus is dyed aubergine (or blue), has a colorless and transparent pod membrane area outside thallus, background is
Black, thallus, background, pod membrane comparison clearly, are judged to the positive；
17F type streptococcus pneumonia thallus is dyed aubergine (or blue), can not see transparent pod membrane area, black outside thallus
Background is connected with thallus, as capsular swelling negative control.
Fig. 5-7 is the 17F type S. pneumoniae capsular swelling negative staining positive and negative photo.
Embodiment 4,19F type S. pneumoniae capsular swelling colouring method
1, material
19F type culture of streptococcus pneumonia liquid, 19F type streptococcus pneumonia antiserum (factor serum 19b), ammonium oxalate
Crystal violet dye liquor, methylene blue dye liquor, sodium chloride injection, 10% prepared Chinese ink
2, method
1) 2 glass slides are taken, 5 μ l of 19F type culture of streptococcus pneumonia liquid is added dropwise respectively；
2) 19F type streptococcus pneumonia antiserum (factor serum 15h) 5 μ l and 5 μ l of sodium chloride injection are added dropwise respectively
It mixes, is sufficiently stirred evenly using micropipettor pipette tips respectively, and drop is tiled with bacterium solution, be placed at room temperature for reaction, naturally dry,
(non-scald on hand is advisable) is fixed through flame；
3) it dyes, ammonium oxalate crystal violet dye liquor is added dropwise or methylenum careuleum dye liquor dyes 1 minute, washing, blotting paper suck dry moisture；
4) negative staining draws a drop prepared Chinese ink and is added drop-wise to glass slide side, tilts specimen slide, contacted with another piece of glass slide
Prepared Chinese ink and with specimen slide at 45 degree of angles, oliquely downward drawing gently, so that prepared Chinese ink covering coloring face, last room temperature is naturally dry
It is dry.
5) microscopy, oil mirror observation.
3, result
19F type streptococcus pneumonia thallus is dyed aubergine (or blue), has a colorless and transparent pod membrane area outside thallus, background is
Black, thallus, background, pod membrane comparison clearly, are judged to the positive；
19F type streptococcus pneumonia thallus is dyed aubergine (or blue), can not see transparent pod membrane area, black outside thallus
Background is connected with thallus, as capsular swelling negative control.
Fig. 8-9 is the 19F type S. pneumoniae capsular swelling negative staining positive and negative photo.
Claims (5)
1. a kind of capsular swelling colouring method of streptococcus pneumonia, which comprises the following steps:
1) 2 glass slides are taken, culture of streptococcus pneumonia liquid is added dropwise respectively；
2) corresponding streptococcus pneumonia specific antisera is added dropwise respectively and sodium chloride injection is mixed with bacterium solution, respectively using micro-
Amount pipettor gun head sufficiently stirs evenly, and drop is tiled, and is placed at room temperature for reaction, naturally dry is fixed through flame；
3) it dyes, ammonium oxalate crystal violet dye liquor or methylene blue dye liquor, washing, blotting paper suck dry moisture is added dropwise；
4) negative staining draws a drop prepared Chinese ink and is added drop-wise to glass slide side, tilts specimen slide, contact prepared Chinese ink with another piece of glass slide
And with specimen slide at 45 degree of angles, oliquely downward drawing gently, so that prepared Chinese ink covering coloring face, last natural drying at room temperature；
5) microscopy, oil mirror observation.
2. colouring method according to claim 1, which is characterized in that further include further deterministic process:
Streptococcus pneumonia thallus dyes aubergine or blue, there is a colorless and transparent pod membrane area outside thallus, and background is black, thallus,
Background, pod membrane comparison clearly, are judged to the positive；
Streptococcus pneumonia thallus dyes aubergine or blue, and transparent pod membrane area, black background and thallus can not be seen outside thallus
It is connected, as capsular swelling negative control.
3. colouring method according to claim 1, which is characterized in that wherein, the concentration of sodium chloride injection is 0.9%.
4. colouring method according to claim 1, which is characterized in that wherein, the concentration of prepared Chinese ink is 10%.
5. colouring method according to claim 1, which comprises the following steps:
1) 2 glass slides are taken, 5 μ l of culture of streptococcus pneumonia liquid is added dropwise respectively；
2) 5 μ l of streptococcus pneumonia specific antisera and chloride injection corresponding to different streptococcus pneumonia types are added dropwise respectively
5 μ l of liquid is mixed with bacterium solution, is sufficiently stirred evenly using micropipettor pipette tips respectively, and drop is tiled, and reaction is placed at room temperature for, natural
It dries, is fixed through flame；
3) it dyes, ammonium oxalate crystal violet dye liquor is added dropwise or methylenum careuleum dye liquor dyes 1 minute, washing, blotting paper suck dry moisture；
4) negative staining draws a drop prepared Chinese ink and is added drop-wise to glass slide side, tilts specimen slide, contact prepared Chinese ink with another piece of glass slide
And with specimen slide at 45 degree of angles, oliquely downward drawing gently, so that prepared Chinese ink covering coloring face, last natural drying at room temperature；
5) microscopy, oil mirror observation；
Capsular swelling colouring method of the present invention, further includes further deterministic process:
Streptococcus pneumonia thallus dyes aubergine or blue, there is a colorless and transparent pod membrane area outside thallus, and background is black, thallus,
Background, pod membrane comparison clearly, are judged to the positive；
Streptococcus pneumonia thallus dyes aubergine or blue, and transparent pod membrane area, black background and thallus can not be seen outside thallus
It is connected, as capsular swelling negative control.
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| CN112129610A (en) * | 2020-09-22 | 2020-12-25 | 无锡市第二人民医院 | Bacterial capsule staining method and application thereof |
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|---|---|---|---|---|
| CN112129610A (en) * | 2020-09-22 | 2020-12-25 | 无锡市第二人民医院 | Bacterial capsule staining method and application thereof |
| CN112129610B (en) * | 2020-09-22 | 2021-07-16 | 无锡市第二人民医院 | Bacterial capsule staining method and application thereof |
| CN112763435A (en) * | 2021-04-01 | 2021-05-07 | 爱沐风优(西安)生物科技有限公司 | Quantitative evaluation method for lethality of streptococcus pneumoniae |
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Application publication date: 20191126 |