CN108663250A - A kind of coccid slide sample production method - Google Patents
A kind of coccid slide sample production method Download PDFInfo
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- CN108663250A CN108663250A CN201810457280.7A CN201810457280A CN108663250A CN 108663250 A CN108663250 A CN 108663250A CN 201810457280 A CN201810457280 A CN 201810457280A CN 108663250 A CN108663250 A CN 108663250A
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- coccid
- production method
- slide sample
- distilled water
- slide
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- 241001465977 Coccoidea Species 0.000 title claims abstract description 99
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 21
- WZRZTHMJPHPAMU-UHFFFAOYSA-L disodium;(3e)-3-[(4-amino-3-sulfonatophenyl)-(4-amino-3-sulfophenyl)methylidene]-6-imino-5-methylcyclohexa-1,4-diene-1-sulfonate Chemical compound [Na+].[Na+].C1=C(S([O-])(=O)=O)C(=N)C(C)=CC1=C(C=1C=C(C(N)=CC=1)S([O-])(=O)=O)C1=CC=C(N)C(S(O)(=O)=O)=C1 WZRZTHMJPHPAMU-UHFFFAOYSA-L 0.000 claims abstract description 9
- 230000001476 alcoholic effect Effects 0.000 claims abstract description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 23
- 238000000926 separation method Methods 0.000 claims description 11
- 229960000583 acetic acid Drugs 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 210000002615 epidermis Anatomy 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 4
- 239000012362 glacial acetic acid Substances 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 4
- 238000002372 labelling Methods 0.000 claims description 4
- 239000002184 metal Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 229920001206 natural gum Polymers 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 241001414830 Diaspididae Species 0.000 claims description 3
- 238000002791 soaking Methods 0.000 claims description 2
- 235000016639 Syzygium aromaticum Nutrition 0.000 claims 1
- 244000223014 Syzygium aromaticum Species 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 20
- 238000004040 coloring Methods 0.000 abstract description 8
- 238000001035 drying Methods 0.000 abstract description 5
- 238000005516 engineering process Methods 0.000 abstract description 4
- 239000012472 biological sample Substances 0.000 abstract description 2
- 238000011010 flushing procedure Methods 0.000 abstract 1
- 230000004048 modification Effects 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 4
- 238000007654 immersion Methods 0.000 description 2
- 230000011164 ossification Effects 0.000 description 2
- 230000009897 systematic effect Effects 0.000 description 2
- 241001133184 Colletotrichum agaves Species 0.000 description 1
- 241001468872 Fumagospora capnodioides Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000002845 discoloration Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 230000000266 injurious effect Effects 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000000126 substance Substances 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
- G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
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)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to biological sample manufacture technology field, more particularly to it is dipped in the NaOH solution that mass concentration is 5%~8% after a kind of coccid slide sample production method, including acquisition coccid low-temperature treatment living and carries out sofening treatment；It is dyed with the acid fuchsine solution after flushing, then with using the alcoholic solution that volumetric concentration is 50%, 70%, 100% to impregnate successively, sample drying is made in last caryophyllus oil after impregnating.The production method of the present invention is easy, safe, and coccid slide sample is clear, completely, uniform coloring, quality height.
Description
Technical field
The invention belongs to biological sample manufacture technology field, more particularly to a kind of coccid slide sample production method.
Background technology
Coccid belongs to Semiptera, Coccoidea.According to coccid net statistics, the coccid recorded in the world at present has
More than 8000 kinds；Record coccid has 14 sections 302 to belong to 1155 kinds in China.The most build of coccid is small, life is hidden, often covers on polypide
There are shell or valuable information for systematic to be protected, vitality and reproductive capacity are extremely strong, are easy to outburst and cause disaster.Most types of coccid
It is important agriculture and forestry injurious insect, plant surface suction juice is often clustered in and causes harm, cause die back, growth potential decline or even whole strain withered
Extremely, very harmful.The excreta of coccid induces sooty mould, and branches and leaves surface is covered with the mould layer of black, influences photosynthesis, reduces ornamental
Value and economic value, seriously affect plant strain growth, influence landscape.The important type of some coccids, which has been cited as national quarantine, to be had
Evil biology.China is vast in territory, and weather is various, and coccid type is various, accurately identifies and identify science of the coccid type to coccid
Prevention and control and comprehensive treatment are of great significance.
Because of coccid Peculiar Forms, structure is subtle, antigeny, and often covering on polypide has shell or valuable information for systematic, no
Easily directly observation identification, therefore usually coccid is fabricated to after slide sample and studies under the microscope.Traditional coccid slide
In Slide processing, there are sample not easy coloring, the low success rate of problems of film-making, and often make clarifier using dimethylbenzene,
Among dimethylbenzene is included in 3 class carcinogenic substance inventories by international cancer research institution of the World Health Organization, human body is poisoned it is larger, no
Safety, not environmentally.
Invention content
The production method of the present invention is easy, safe, and coccid slide sample is clear, completely, uniform coloring, quality height.
The present invention provides a kind of coccid slide sample production methods, include the following steps：
S1 acquires coccid living, is stored at -10~-5 DEG C, obtain pending coccid；
Pending coccid pricks 1~2 hole in polypide epidermis in S2, S1, and it is 5%~8% to be then dipped in mass concentration
1~6h in NaOH solution, and squeezing makes coccid polypide inner material dissolve out, and must soften coccid；
S3, distilled water-acetum, which rinses in S2, softens coccid, and washing time >=1h is then placed in the acid fuchsine solution
Middle immersion 2~for 24 hours, coccid must be dyed；
Wherein, distilled water-acetum is prepared in accordance with the following methods：2~3 drop acetic acid are added dropwise in per 10ml distilled water；
In S4, S3 dye coccid successively use volumetric concentration be 50%, 70%, 100% alcoholic solution respectively impregnate 15~
20min must be dehydrated color separation coccid；
It is dehydrated color separation coccid in S5, S4 to be dipped in caryophyllus oil, obtains transparent coccid；
S6, drop 1~2 drop in glass slide center is gummy, and transparent coccid is placed in natural gum in S5, covered, and labelling is right
Drying at room temperature afterwards obtains coccid slide sample.
Preferably, when pending coccid belongs to lecanium or armored scale in S2, impregnate before prior to 45~50 DEG C in metal bath 0.5~
1h。
Preferably, the acid fuchsine solution is made in accordance with the following methods in S3：1g is pinkish red, volumetric concentration is 96% alcohol
10ml, 5ml glacial acetic acid, 100ml distilled water, mixing filter to take filtrate afterwards for 24 hours.
Preferably, it is 15~25min that soaking time of the color separation coccid in caryophyllus oil is dehydrated in S4.
Compared with prior art, beneficial effects of the present invention：
The coccid production method of the present invention carries out kill fixation using low temperature to coccid living, and coccid is made to keep complete, fresh-keeping
Color is protected, coccid drying shrinkage, discoloration or damage are prevented；NaOH solution softening can remove coccid content, body surface secretion, external
Substance squeezes fully penetrating spread apart while polypide makes polypide not be destroyed in softening；It is upper extra that acetic acid effectively neutralizes coccid
NaOH solution, be conducive to follow-up coloring, first dye and be dehydrated polypide easy coloring again；50%, 70%, 100% alcoholic solution is successively
It impregnates, step is simplified and dehydration color separation is carried out at the same time, and prevents polypide shrinkage and embrittlement；It is safe and non-toxic that caryophyllus oil makees clarifier.This
Invention is easy, safety, and coccid slide sample is clear, and completely, uniform coloring, quality is high, convenient for clear observation under the microscope and
Identification.
Description of the drawings
Fig. 1 is the schematic diagram of coccid slide of the present invention under the microscope.
Specific implementation mode
1 pair of several specific implementation mode of the invention is described in detail below in conjunction with the accompanying drawings, it is to be understood that of the invention
Protection domain be not restricted by specific implementation.
Embodiment 1
A kind of coccid slide sample production method, includes the following steps：
S1 acquires coccid living, is stored at -10 DEG C, obtain pending coccid；
Pending coccid pricks 1 hole in polypide epidermis in S2, S1, is then dipped in the NaOH solution that mass concentration is 5%
1h, and squeezing makes coccid polypide inner material dissolve out, and retains coccid cage, must soften coccid；
If pending coccid belongs to the higher lecanium of ossification intensity in S2, before impregnating prior to 45 DEG C in metal bath 0.5h；
S3, distilled water-acetum, which rinses in S2, softens coccid, and washing time >=1h is then placed in the acid fuchsine solution
Middle immersion 2h, must dye coccid；
Wherein, distilled water-acetum is prepared in accordance with the following methods：2~3 drop acetic acid are added dropwise in per 10ml distilled water；
The acid fuchsine solution is made in accordance with the following methods：1g is pinkish red, volumetric concentration is 96% alcohol 10ml, 5ml glacial acetic acid,
100ml distilled water, mixing filter to take filtrate afterwards for 24 hours；
The alcoholic solution that coccid is successively 50%, 70%, 100% with volumetric concentration is dyed in S4, S3 respectively impregnates 15min,
Color separation coccid must be dehydrated；
It is dehydrated color separation coccid in S5, S4 and is dipped in 15min in caryophyllus oil, obtains transparent coccid；
S6, drop 1~2 drop in glass slide center is gummy, and transparent coccid is placed in natural gum in S5, covered, and labelling is right
Drying at room temperature afterwards obtains coccid slide sample.
The micrograph of coccid slide sample obtained is as shown in Figure 1, sample is clear, and completely, uniform coloring, quality is high, just
In clear observation and identification.
Embodiment 2
A kind of coccid slide sample production method, includes the following steps：
S1 acquires coccid living, is stored at -5 DEG C, obtain pending coccid；
Pending coccid pricks 2 holes in polypide epidermis in S2, S1, is then dipped in the NaOH solution that mass concentration is 8%
6h, and squeezing makes coccid polypide inner material dissolve out, and retains coccid cage, must soften coccid；
If pending coccid belongs to the higher armored scale of ossification intensity in S2, before impregnating prior to 50 DEG C in metal bath 1h；
S3, distillation-acetum water, which rinses in S2, softens coccid, and washing time >=1h is then placed in the acid fuchsine solution
For 24 hours, coccid must be dyed；
Wherein, distilled water-acetum is prepared in accordance with the following methods：2~3 drop acetic acid are added dropwise in per 10ml distilled water；
The acid fuchsine solution is made in accordance with the following methods：1g is pinkish red, volumetric concentration is 96% alcohol 10ml, 5ml glacial acetic acid,
100ml distilled water, mixing filter to take filtrate afterwards for 24 hours；
The alcoholic solution that coccid is successively 50%, 70%, 100% with volumetric concentration is dyed in S4, S3 respectively impregnates 20min,
Color separation coccid must be dehydrated；
It is dehydrated color separation coccid in S5, S4 and is dipped in 25min in caryophyllus oil, obtains transparent coccid；
S6, drop 1~2 drop in glass slide center is gummy, and transparent coccid is placed in natural gum in S5, covered, and labelling is right
Drying at room temperature afterwards obtains coccid slide sample.
It is made sample 200, each sample is clear, complete, uniform coloring, subsequently to be provided very to the research of coccid
Good material sample.
It should be noted that the step method used in claims of the present invention is same as the previously described embodiments, in order to anti-
It only repeats, description of the invention preferred embodiment, once a person skilled in the art knows basic creative general
It reads, then additional changes and modifications can be made to these embodiments.So it includes preferred real that the following claims are intended to be interpreted as
It applies example and falls into all change and modification of the scope of the invention.
Obviously, various changes and modifications can be made to the invention without departing from essence of the invention by those skilled in the art
God and range.In this way, if these modifications and changes of the present invention belongs to the range of the claims in the present invention and its equivalent technologies
Within, then the present invention is also intended to include these modifications and variations.
Claims (4)
1. a kind of coccid slide sample production method, which is characterized in that include the following steps：
S1 acquires coccid living, is stored at -10~-5 DEG C, obtain pending coccid；
Pending coccid pricks 1~2 hole in polypide epidermis in S2, S1, and it is molten to be then dipped in the NaOH that mass concentration is 5%~8%
1~6h in liquid, and squeezing makes coccid polypide inner material dissolve out, and must soften coccid；
S3, distilled water-acetum, which rinses in S2, softens coccid, and washing time >=1h is then placed in the acid fuchsine solution and soaks
Bubble 2~for 24 hours, coccid must be dyed；
Wherein, distilled water-acetum is prepared in accordance with the following methods：2~3 drop acetic acid are added dropwise in per 10ml distilled water；
Coccid is dyed in S4, S3 uses the alcoholic solution that volumetric concentration is 50%, 70%, 100% respectively to impregnate 15~20min successively,
Color separation coccid must be dehydrated；
It is dehydrated color separation coccid in S5, S4 to be dipped in caryophyllus oil, obtains transparent coccid；
S6, drop 1~2 drop in glass slide center is gummy, and transparent coccid is placed in natural gum in S5, covered, labelling and then room
Temperature is dry, obtains coccid slide sample.
2. coccid slide sample production method as described in claim 1, which is characterized in that pending coccid belongs to lecanium in S2
Or when armored scale, impregnate before prior to 45~50 DEG C in 0.5~1h of metal bath.
3. coccid slide sample production method as described in claim 1, which is characterized in that in S3 the acid fuchsine solution according to
Lower section method is made：1g is pinkish red, volumetric concentration is 96% alcohol 10ml, 5ml glacial acetic acid, 100ml distilled water, and mixing filters afterwards for 24 hours
Take filtrate.
4. coccid slide sample production method as described in claim 1, which is characterized in that be dehydrated color separation coccid in S4 in cloves
Soaking time in oil is 15~25min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810457280.7A CN108663250A (en) | 2018-05-14 | 2018-05-14 | A kind of coccid slide sample production method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810457280.7A CN108663250A (en) | 2018-05-14 | 2018-05-14 | A kind of coccid slide sample production method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108663250A true CN108663250A (en) | 2018-10-16 |
Family
ID=63779414
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810457280.7A Pending CN108663250A (en) | 2018-05-14 | 2018-05-14 | A kind of coccid slide sample production method |
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| Country | Link |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109580300A (en) * | 2018-11-06 | 2019-04-05 | 无锡市儿童医院 | The method and its application of dust mite in a kind of collection dust |
| CN109827832A (en) * | 2019-01-17 | 2019-05-31 | 河南师范大学 | The production method of branch angular motion object Permanent slide |
| CN112889807A (en) * | 2021-02-09 | 2021-06-04 | 陕西理工大学 | Method for manufacturing permanent microscopic slide specimen of tiny insect |
| CN113155563A (en) * | 2021-04-13 | 2021-07-23 | 凭祥海关综合技术服务中心 | Method for preparing scale insect specimen |
| CN113575569A (en) * | 2021-05-24 | 2021-11-02 | 广东生态工程职业学院 | Method for dissolving ceroplasmic substance of Lecanidae coccid |
| CN114199644A (en) * | 2021-11-23 | 2022-03-18 | 凭祥海关综合技术服务中心 | Rapid manufacturing method of orographic interception mealybug quarantine identification slide |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109580300A (en) * | 2018-11-06 | 2019-04-05 | 无锡市儿童医院 | The method and its application of dust mite in a kind of collection dust |
| CN109580300B (en) * | 2018-11-06 | 2021-09-24 | 无锡市儿童医院 | Method for collecting dust mites in dust and application thereof |
| CN109827832A (en) * | 2019-01-17 | 2019-05-31 | 河南师范大学 | The production method of branch angular motion object Permanent slide |
| CN112889807A (en) * | 2021-02-09 | 2021-06-04 | 陕西理工大学 | Method for manufacturing permanent microscopic slide specimen of tiny insect |
| CN113155563A (en) * | 2021-04-13 | 2021-07-23 | 凭祥海关综合技术服务中心 | Method for preparing scale insect specimen |
| CN113575569A (en) * | 2021-05-24 | 2021-11-02 | 广东生态工程职业学院 | Method for dissolving ceroplasmic substance of Lecanidae coccid |
| CN114199644A (en) * | 2021-11-23 | 2022-03-18 | 凭祥海关综合技术服务中心 | Rapid manufacturing method of orographic interception mealybug quarantine identification slide |
| CN114199644B (en) * | 2021-11-23 | 2024-04-26 | 凭祥海关综合技术服务中心 | Quick manufacturing method of port interception mealybugs quarantine identification slide |
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Application publication date: 20181016 |