CN114235524A - Sputum treatment method - Google Patents
Sputum treatment method Download PDFInfo
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- CN114235524A CN114235524A CN202111505279.5A CN202111505279A CN114235524A CN 114235524 A CN114235524 A CN 114235524A CN 202111505279 A CN202111505279 A CN 202111505279A CN 114235524 A CN114235524 A CN 114235524A
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- sputum
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- mixing kettle
- preparing
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- 206010036790 Productive cough Diseases 0.000 title claims abstract description 44
- 210000003802 sputum Anatomy 0.000 title claims abstract description 44
- 208000024794 sputum Diseases 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 19
- 238000002156 mixing Methods 0.000 claims abstract description 41
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 34
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 34
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims abstract description 26
- 239000011780 sodium chloride Substances 0.000 claims abstract description 17
- 102000004142 Trypsin Human genes 0.000 claims abstract description 16
- 108090000631 Trypsin Proteins 0.000 claims abstract description 16
- 239000012588 trypsin Substances 0.000 claims abstract description 16
- 239000006081 fluorescent whitening agent Substances 0.000 claims abstract description 12
- 239000003906 humectant Substances 0.000 claims abstract description 12
- 239000003961 penetration enhancing agent Substances 0.000 claims abstract description 12
- 238000005303 weighing Methods 0.000 claims abstract description 12
- 238000003756 stirring Methods 0.000 claims abstract description 11
- 238000004043 dyeing Methods 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims description 31
- 239000011521 glass Substances 0.000 claims description 8
- 239000012192 staining solution Substances 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 238000010186 staining Methods 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 241000233866 Fungi Species 0.000 abstract description 9
- 238000001514 detection method Methods 0.000 abstract description 6
- 239000012530 fluid Substances 0.000 abstract description 4
- 201000010099 disease Diseases 0.000 abstract description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 239000010813 municipal solid waste Substances 0.000 abstract description 3
- 230000005484 gravity Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 101710131583 Trypsin-10 Proteins 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 230000001338 necrotic effect Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 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
-
- 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
- G01N1/38—Diluting, dispersing or mixing samples
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the technical field of fungus detection, in particular to a sputum treatment method, which aims at solving the problem that when a patient in an existing hospital treats sputum, the sputum is generally taken in a garbage can to cause some fungi to spread everywhere, so that various diseases are spread, and the following scheme is proposed, and comprises the following steps: s1: preparing a mixing kettle, a centrifuge, an oven and a weighing instrument; s2: adding trypsin, KOH, sodium chloride and ethanol into a mixing kettle, mixing and stirring uniformly, and preparing a treatment solution; s3: adding the fluorescent whitening agent, the penetration enhancer, the humectant and the background counterstain into a mixing kettle, mixing and stirring uniformly, and preparing a dyeing solution; s4: and placing the sputum sample in the treatment fluid according to different proportions, and centrifuging by using a centrifuge. The method is simple to operate, can reduce the biological infectivity of the sputum sample, can improve the detection rate of fungi, and is convenient for people to use.
Description
Technical Field
The invention relates to the technical field of fungus detection, in particular to a method for treating sputum.
Background
Sputum is liquid secreted by respiratory tract under stimulation, the components of the sputum comprise mucus, foreign matters, pathogenic microorganisms, various inflammatory cells, necrotic and exfoliated mucosal epithelial cells and the like, and some hospital liquid can be used for detecting the sputum of patients so as to obtain different symptoms.
When a patient in an existing hospital treats sputum, the patient generally hurts the sputum into a trash can, so that some fungi are spread everywhere, and various diseases are spread, and therefore a method for treating the sputum is provided.
Disclosure of Invention
The invention aims to solve the defect that when a patient in an existing hospital treats sputum, the sputum is generally sent into a garbage can to cause fungi to spread everywhere, so that various diseases are spread, and provides a method for treating the sputum.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for treating sputum comprises the following steps:
s1: preparing a mixing kettle, a centrifuge, an oven and a weighing instrument;
s2: adding trypsin, KOH, sodium chloride and ethanol into a mixing kettle, mixing and stirring uniformly, and preparing a treatment solution;
s3: adding the fluorescent whitening agent, the penetration enhancer, the humectant and the background counterstain into a mixing kettle, mixing and stirring uniformly, and preparing a dyeing solution;
s4: placing the sputum sample in different proportions in a treatment solution, centrifuging by a centrifuge,
s5: and dripping the centrifuged sample on a glass slide, drying by a dryer, adding a dyeing solution, covering a cover and breaking the slide, and observing by using a microscope.
Preferably, in the S2, trypsin, KOH, sodium chloride and ethanol are quantitatively determined by a weighing machine, 10 to 15 parts of trypsin, 7 to 10 parts of KOH, 5 to 7 parts of sodium chloride and 5 to 6 parts of ethanol are quantitatively determined, and the mixture is stirred for 5 minutes by a mixing kettle.
Preferably, in S2, a weighing apparatus is used to quantify the fluorescent whitening agent, the penetration enhancer, the humectant and the background counterstain, wherein 8 to 10 parts of the fluorescent whitening agent, 5 to 7 parts of the penetration enhancer, 3 to 5 parts of the humectant and 2 to 3 parts of the background counterstain are stirred in a mixing kettle for 3 minutes.
Preferably, the sputum is placed in the treatment solution in the step S4 and treated by a centrifuge.
Preferably, the treatment solutions for sputum with different concentrations in S5 are respectively placed on glass slides, dried by a dryer, treated by a staining solution for 2 minutes, and then respectively observed microscopically by a microscope.
Preferably, the rotating speed of the mixing kettle is 150 revolutions per minute, and a timer is arranged on the mixing kettle.
Preferably, the trypsin, KOH, sodium chloride and ethanol are all purified by filtration prior to use.
Preferably, the sputum is aliquoted separately from the treatment fluid at 1/5, 1/10 and 1/20.
Compared with the prior art, the invention has the advantages that:
(1) the technical scheme is characterized in that the treatment solution is prepared according to a certain proportion, and the sputum sample is digested, so that the biological infectivity of the sputum sample is reduced after digestion, and the detection rate of fungi can be improved;
(2) the fluorescent staining solution is adopted to detect the fungi, the operation is simple, the time consumption is short, the detection rate is high, and the application is less clinically at present.
The method is simple to operate, can reduce the biological infectivity of the sputum sample, can improve the detection rate of fungi, and is convenient for people to use.
Detailed Description
The technical solutions in the embodiments will be described clearly and completely below, and it is obvious that the described embodiments are only a part of the embodiments, but not all embodiments.
Example one
A method for treating sputum comprises the following steps:
s1: preparing a mixing kettle, a centrifuge, an oven and a weighing instrument;
s2: adding trypsin, KOH, sodium chloride and ethanol into a mixing kettle, mixing and stirring uniformly, and preparing a treatment solution;
s3: adding the fluorescent whitening agent, the penetration enhancer, the humectant and the background counterstain into a mixing kettle, mixing and stirring uniformly, and preparing a dyeing solution;
s4: placing the sputum samples in different proportions in a treatment solution, and centrifuging by a centrifuge;
s5: and dripping the centrifuged sample on a glass slide, drying by a dryer, adding a dyeing solution, covering a cover and breaking the slide, and observing by using a microscope.
In this example, in S2, trypsin, KOH, sodium chloride, and ethanol were quantitatively determined using a weighing apparatus, and trypsin 10 to 15 parts, KOH7 to 10 parts, sodium chloride 5 to 7 parts, and ethanol 5 to 6 parts were stirred in a mixing tank for 5 minutes.
In this embodiment, in S2, a weighing apparatus is used to quantify the fluorescent whitening agent, the penetration enhancer, the humectant, and the background counterstain, wherein 8 to 10 parts of the fluorescent whitening agent, 5 to 7 parts of the penetration enhancer, 3 to 5 parts of the humectant, and 2 to 3 parts of the background counterstain are stirred in a mixing kettle for 3 minutes.
In this embodiment, the sputum is placed in the processing solution and processed by a centrifuge in S4.
In this embodiment, the treatment solutions for sputum with different concentrations in S5 are respectively placed on a glass slide, dried by a dryer, treated with a staining solution for 2 minutes, and then respectively observed under a microscope.
In this embodiment, the rotation speed of the mixing kettle is 150 rpm, and a timer is arranged on the mixing kettle.
In this example, the trypsin, KOH, sodium chloride and ethanol were all purified by filtration prior to use.
In this example, the sputum was aliquoted separately from the treatment fluid at 1/5, 1/10 and 1/20.
Example two
A method for treating sputum comprises the following steps:
s1: preparing a mixing kettle, a centrifuge, an oven and a weighing instrument;
s2: adding trypsin, KOH, sodium chloride and ethanol into a mixing kettle, mixing and stirring uniformly, and preparing a treatment solution;
s3: adding the fluorescent whitening agent, the penetration enhancer, the humectant and the background counterstain into a mixing kettle, mixing and stirring uniformly, and preparing a dyeing solution;
s4: placing the sputum sample in different proportions in a treatment solution, centrifuging by a centrifuge,
s5: and dripping the centrifuged sample on a glass slide, drying by a dryer, adding a dyeing solution, covering a cover and breaking the slide, and observing by using a microscope.
In this embodiment, in S2, a weighing apparatus is used to quantify trypsin, KOH, sodium chloride, and ethanol, 10 to 15 parts of trypsin, 7 to 10 parts of KOH, 5 to 7 parts of sodium chloride, and 5 to 6 parts of ethanol, and the mixture is stirred in a mixing kettle for 5 minutes, so that different treatment solutions can be obtained through materials with different specific gravities, and the purpose of obtaining the optimal digested sputum can be achieved through experiments.
In this embodiment, in S2, a weighing apparatus is used to quantify the fluorescent whitening agent, the penetration enhancer, the humectant and the background counterstain, wherein 8 to 10 parts of the fluorescent whitening agent, 5 to 7 parts of the penetration enhancer, 3 to 5 parts of the humectant and 2 to 3 parts of the background counterstain are stirred in a mixing kettle for 3 minutes, and different staining solutions can be obtained through materials with different specific gravities, so that the treated sputum can be observed more obviously when being stained.
In this embodiment, place the sputum in the processing fluid among the S4 to handle with centrifuge, pour the supernatant with the sample after the centrifugation, leave about 1 mL' S bottom solution for use, thereby can detect more accurately.
In this embodiment, the treatment solutions to the sputum with different concentrations in S5 are respectively placed on the glass slide, dried by the dryer, and treated by the staining solution for 2 minutes, and then respectively observed microscopically by the microscope, and the staining solution is firstly used for staining, and then observed and detected by the microscope.
In this embodiment, the rotational speed of mixing cauldron is 150 revolutions per minute, and is equipped with the timer on the mixing cauldron, and through the timer timing when the mixing cauldron stirs.
In this example, the trypsin, KOH, sodium chloride and ethanol were all purified by filtration before use, thereby providing more thorough treatment of the sputum.
In this embodiment, the sputum is placed in equal parts with the treatment solution according to 1/5, 1/10, and 1/20, respectively, so that the treatment effect can be observed at different treatment concentrations.
The above description is only a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered as the technical solutions and the inventive concepts of the present invention in the technical scope of the present invention.
Claims (8)
1. A method for treating sputum, comprising the steps of:
s1: preparing a mixing kettle, a centrifuge, an oven and a weighing instrument;
s2: adding trypsin, KOH, sodium chloride and ethanol into a mixing kettle, mixing and stirring uniformly, and preparing a treatment solution;
s3: adding the fluorescent whitening agent, the penetration enhancer, the humectant and the background counterstain into a mixing kettle, mixing and stirring uniformly, and preparing a dyeing solution;
s4: placing the sputum sample in different proportions in a treatment solution, centrifuging by a centrifuge,
s5: and dripping the centrifuged sample on a glass slide, drying by a dryer, adding a dyeing solution, covering a cover and breaking the slide, and observing by using a microscope.
2. The method for processing sputum according to claim 1, wherein trypsin, KOH, sodium chloride and ethanol are quantitatively determined in S2 by using a weighing apparatus, and trypsin 10-15 parts, KOH7-10 parts, sodium chloride 5-7 parts and ethanol 5-6 parts are stirred for 5 minutes through a mixing kettle.
3. The method according to claim 1, wherein 8 to 10 parts of fluorescent whitening agent, 5 to 7 parts of penetration enhancer, 3 to 5 parts of humectant and 2 to 3 parts of background counter-staining agent are weighed in S2 and stirred in a mixing kettle for 3 minutes.
4. The method according to claim 1, wherein the sputum is placed in a treatment solution and centrifuged in S4.
5. The method according to claim 1, wherein the sputum treatment solutions with different concentrations in S5 are separately placed on a slide glass, dried by a dryer, and treated with a staining solution for 2 minutes, and then observed microscopically by a microscope.
6. The method according to claim 2, wherein the mixing kettle rotates at a speed of 150 rpm and is provided with a timer.
7. The method of claim 1, wherein the trypsin, KOH, NaCl and ethanol are purified by filtration before use.
8. The method of claim 1, wherein the sputum is aliquoted from the treatment solution at 1/5, 1/10 and 1/20.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111505279.5A CN114235524B (en) | 2021-12-10 | 2021-12-10 | Sputum treatment method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111505279.5A CN114235524B (en) | 2021-12-10 | 2021-12-10 | Sputum treatment method |
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| CN114235524A true CN114235524A (en) | 2022-03-25 |
| CN114235524B CN114235524B (en) | 2024-02-23 |
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