CN109459262A - A kind of preparation method that maturation maize root system is temporarily sliced - Google Patents
A kind of preparation method that maturation maize root system is temporarily sliced Download PDFInfo
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- CN109459262A CN109459262A CN201811570886.8A CN201811570886A CN109459262A CN 109459262 A CN109459262 A CN 109459262A CN 201811570886 A CN201811570886 A CN 201811570886A CN 109459262 A CN109459262 A CN 109459262A
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- preparation
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- root system
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- 240000008042 Zea mays Species 0.000 title claims abstract description 32
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 32
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 title claims abstract description 28
- 235000009973 maize Nutrition 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 230000035800 maturation Effects 0.000 title claims abstract description 11
- 238000004043 dyeing Methods 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 239000011521 glass Substances 0.000 claims description 13
- 239000000463 material Substances 0.000 claims description 11
- 239000012153 distilled water Substances 0.000 claims description 10
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 4
- 235000005822 corn Nutrition 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 4
- 239000000975 dye Substances 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 5
- 238000000034 method Methods 0.000 abstract description 4
- 210000003484 anatomy Anatomy 0.000 abstract description 2
- 238000011160 research Methods 0.000 abstract description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- 239000008267 milk Substances 0.000 description 4
- 210000004080 milk Anatomy 0.000 description 4
- 235000013336 milk Nutrition 0.000 description 4
- 238000000879 optical micrograph Methods 0.000 description 4
- 238000011010 flushing procedure Methods 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 238000013459 approach Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 230000005070 ripening Effects 0.000 description 1
- 238000012360 testing method Methods 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
- G01N1/02—Devices for withdrawing samples
- G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
- G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
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- 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)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
A kind of preparation method that maturation maize root system is temporarily sliced belongs to root system of plant slice preparation technical field.The present invention is for fogging image in existing interim slice preparation process, to lead to the problem of the error of observation, provide a kind of preparation method that mature maize root system is temporarily sliced, slice after dyeing is subjected to vacuumize process, exclude in slice 99% bubble, it is more clear observation image, reduces the error of observation, the research for mature maize root system anatomical structure is provided convenience.
Description
Technical field
The invention belongs to root systems of plant to be sliced preparation technical field, and in particular to a kind of maturation maize root system is temporarily sliced
Preparation method.
Background technique
Interim slice is used for observation of plant internal structure, but finds in actual experiment, and the interim conventional method that is sliced is used for
More tender plant tissue has preferable effect, but when plant comes to the ripening period, especially maize root system, cell wall add
Thickness, after slice, when observation, it may appear that a large amount of bubbles influence to observe result.
Summary of the invention
For the problem that fogging image in existing interim slice preparation process, to generate the error of observation, the present invention is provided
A kind of preparation method that mature maize root system is temporarily sliced, includes the following steps:
1) the corn root cross section in maturity period is scabbled, block of cutting into chunks, it is fixed in FAA fixer or Ka Nuoshi liquid, Gu
It fixes time as 20-72h；
2) it then, takes out material and cuts thin slice, thin slice is placed in water；
3) thin and transparent slice is selected, after dyeing, is placed on glass slide；
4) distilled water then, is added dropwise on tangential section, after being vacuum-treated, covered suits slice with glass slide
Afterwards, mature maize root system is prepared temporarily to be sliced.
Further limit, described section of block of step 1) with a thickness of 1-2cm.
It further limits, the step 1) set time is for 24 hours.
It further limits, the step 3) dyeing refers to that the sarranine aqueous solution by slice through mass fraction 1% dyes 20-
40s, preferably 30s.
It further limits, distilled water is added dropwise on the step 4) tangential section, the amount of dropwise addition is 120-150 μ L.
It further limits, the step 4) vacuum processing refers to slice under the conditions of vacuum degree is 5-10torr, takes out
Vacuum 5-10min.
It further limits, the step 4) vacuum processing refers to slice under the conditions of vacuum degree is 8torr, takes out true
Empty 7min.
Beneficial effect
The present invention uses vacuum pump, and the slice after dyeing is carried out vacuumize process, can exclude in slice 99% bubble,
It is more clear observation image, reduces the error of observation, saves a large amount of time, for the research of mature maize root system anatomical structure
It provides convenience.
Detailed description of the invention
Fig. 1 maturation maize root system is sliced the image through 40 times of optical microphotograph sem observations after dyeing.
After Fig. 2 maturation maize root system slice is dyed, is vacuumized, the image through 40 times of optical microphotograph sem observations.
Fig. 3 maturation maize root system is sliced the image through 100 times of optical microphotograph sem observations after dyeing.
After Fig. 4 maturation maize root system slice is dyed, is vacuumized, the image through 100 times of optical microphotograph sem observations.
Specific embodiment
In following embodiments, the mature maize root system sample is the root system for being derived from corn milk stage.
FAA fixer used is called standard fixer, by formalin 5mL, glacial acetic acid 5mL, 70% second of volume fraction
It is formulated after alcohol 90mL, glycerine 5mL mixing.
Ka Nuoshi liquid: acquisition is mixed with according to 3:1 volume ratio by dehydrated alcohol and glacial acetic acid.
Other reagents or instrument etc. unless otherwise specified, can be bought by commercialization approach and be obtained.
The preparation method that the mature maize root system of embodiment 1. is temporarily sliced.
1) the milk stage maize root system material distilled water repeated flushing fetched is clean, material cross section is scabbled, is cut
At the section block of 1-2cm, fixed for 24 hours in FAA fixer (standard fixer).
2) double-edged razor blade is selected after taking out material, with uniform strength, makes blade planar stable sliding row, cuts thin slice and gently move into
It is spare in the culture dish being filled with water.
3) thin and transparent slice is selected, is advisable with a thickness of 50~100 μm, is contaminated with the sarranine aqueous solution of mass fraction 1%
Color 30s is subsequently placed in glass slide center.30s is the optimum dyeing time, and overlong time be easy to cause and overstains, and influences image
Observation.
4) 3-4 drop distilled water is dripped on tangential section, 120 μ L are advisable, then slice is put into vacuum pump, are in vacuum degree
Under the conditions of 8torr, 7min (slice be bonded completely with glass slide until) is vacuumized, (this step must be for covered after taking-up
After vacuumizing, the positive effect for excluding bubble is otherwise not achieved), with tweezers gently gland slide, keep slice tight with glass slide
Closely connected conjunction is made mature maize root system and is temporarily sliced, observes under the microscope.
The preparation method that the mature maize root system of embodiment 2. is temporarily sliced.
1) the milk stage maize root system material distilled water repeated flushing fetched is clean, material cross section is scabbled, is cut
At the section block of 1-2cm, 20h is fixed in FAA fixer (standard fixer).
2) double-edged razor blade is selected after taking out material, with uniform strength, makes blade planar stable sliding row, cuts thin slice and gently move into
It is spare in the culture dish being filled with water.
3) thin and transparent slice is selected, is advisable with a thickness of 50~100 μm, is contaminated with the sarranine aqueous solution of mass fraction 1%
Color 20s is subsequently placed in glass slide center.
4) 3-4 drop distilled water is dripped on tangential section, 150 μ L are advisable, then slice is put into vacuum pump, are in vacuum degree
Under the conditions of 5torr, vacuumize 5min (until slice is bonded completely with glass slide), covered after taking-up, gently with tweezers
Gland slide fits closely slice with glass slide, mature maize root system is made and is temporarily sliced.
The preparation method that the mature maize root system of embodiment 3. is temporarily sliced.
1) the milk stage maize root system material distilled water repeated flushing fetched is clean, material cross section is scabbled, is cut
At the section block of 1-2cm, 72h is fixed in Ka Nuoshi liquid.
2) double-edged razor blade is selected after taking out material, with uniform strength, makes blade planar stable sliding row, cuts thin slice and gently move into
It is spare in the culture dish being filled with water.
3) thin and transparent slice is selected, is advisable with a thickness of 50~100 μm, is contaminated with the sarranine aqueous solution of mass fraction 1%
Color 40s is subsequently placed in glass slide center.
4) 3-4 drop distilled water is dripped on tangential section, 130 μ L are advisable, then slice is put into vacuum pump, are in vacuum degree
Under the conditions of 10torr, 10min (until slice is bonded completely with glass slide) is vacuumized, covered after taking-up is light with tweezers
Light cap slide fits closely slice with glass slide, mature maize root system is made and is temporarily sliced.
The preparation method that 4. maize root system of embodiment is temporarily sliced.
Repeat embodiment 1, be with the difference of embodiment 1, in the present embodiment in step 1) root system sample in FAA fixer
48h is fixed in (standard fixer).
For the maturation maize root system made of the embodiment 1 is temporarily sliced, slice is sliced after being made through micro- sem observation, such as
Shown in Fig. 2 and Fig. 4, after slice dyeing after vacuumize process, compared to not vacuum-treated slice (as shown in figures 1 and 3),
Bubble (black real point) almost all is excluded in the visible corn matured root biopsy tissues of microscopically observation, and observation image is more
Clearly, reduce the error of observation, keep test result more acurrate.
Claims (8)
1. a kind of preparation method that maturation maize root system is temporarily sliced, which comprises the steps of:
1) the corn root cross section in maturity period is scabbled, block of cutting into chunks, it is fixed in FAA fixer or Ka Nuoshi liquid, when fixed
Between be 20-72h；
2) it then, takes out material and cuts thin slice, thin slice is placed in water；
3) thin and transparent slice is selected, after dyeing, is placed on glass slide；
4) distilled water then, is added dropwise on tangential section, after being vacuum-treated, covered, after suiting slice with glass slide, system
The standby maturation maize root system that obtains temporarily is sliced.
2. preparation method according to claim 1, which is characterized in that described section of block of step 1) with a thickness of 1-2cm.
3. preparation method according to claim 1, which is characterized in that the step 1) set time is for 24 hours.
4. preparation method according to claim 1, which is characterized in that the step 3) dyeing refers to slice through quality point
The sarranine aqueous solution of number 1% dyes 20-40s.
5. the preparation method according to claim 4, which is characterized in that the step 3) dyeing time is 30s.
6. preparation method according to claim 1, which is characterized in that distilled water is added dropwise on the step 4) tangential section, drips
The amount added is 120-150 μ L.
7. preparation method according to claim 1, which is characterized in that the step 4) vacuum processing refers to and will be sliced true
Under the conditions of reciprocal of duty cycle is 5-10torr, 5-10min is vacuumized.
8. preparation method according to claim 7, which is characterized in that the step 4) vacuum processing refers to and will be sliced true
Under the conditions of reciprocal of duty cycle is 8torr, 7min is vacuumized.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811570886.8A CN109459262A (en) | 2018-12-21 | 2018-12-21 | A kind of preparation method that maturation maize root system is temporarily sliced |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811570886.8A CN109459262A (en) | 2018-12-21 | 2018-12-21 | A kind of preparation method that maturation maize root system is temporarily sliced |
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| Publication Number | Publication Date |
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| CN109459262A true CN109459262A (en) | 2019-03-12 |
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| CN201811570886.8A Pending CN109459262A (en) | 2018-12-21 | 2018-12-21 | A kind of preparation method that maturation maize root system is temporarily sliced |
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| Country | Link |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110646257A (en) * | 2019-08-27 | 2020-01-03 | 中国科学院东北地理与农业生态研究所 | Flaking method for observing aerial root microstructure of corn and application thereof |
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| CN103149189A (en) * | 2013-03-11 | 2013-06-12 | 山东省农业科学院作物研究所 | Method for rapidly detecting wheat tissue callose |
| CN104132940A (en) * | 2014-06-30 | 2014-11-05 | 上海市园林科学研究所 | Convenient observation method of orchid mycorrhiza microstructure |
| CN106404475A (en) * | 2016-06-29 | 2017-02-15 | 西南大学 | A method of preparing a dactylis glomerata root tissue paraffin section |
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| CN107167350A (en) * | 2017-05-16 | 2017-09-15 | 云南省农业科学院园艺作物研究所 | A kind of preparation method of the paraffin section of eggplant rhizome portion tissue |
-
2018
- 2018-12-21 CN CN201811570886.8A patent/CN109459262A/en active Pending
Patent Citations (6)
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| WO2000020846A1 (en) * | 1998-10-02 | 2000-04-13 | Rogers Imaging Corporation | Method for imaging tissue |
| CN103149189A (en) * | 2013-03-11 | 2013-06-12 | 山东省农业科学院作物研究所 | Method for rapidly detecting wheat tissue callose |
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| Title |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110646257A (en) * | 2019-08-27 | 2020-01-03 | 中国科学院东北地理与农业生态研究所 | Flaking method for observing aerial root microstructure of corn and application thereof |
| CN110646257B (en) * | 2019-08-27 | 2022-05-24 | 中国科学院东北地理与农业生态研究所 | Flaking method for observing aerial root microstructure of corn and application thereof |
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Application publication date: 20190312 |