CN113295490A - Method for making paraffin section of cedrela sinensis callus - Google Patents
Method for making paraffin section of cedrela sinensis callus Download PDFInfo
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- CN113295490A CN113295490A CN202110559019.XA CN202110559019A CN113295490A CN 113295490 A CN113295490 A CN 113295490A CN 202110559019 A CN202110559019 A CN 202110559019A CN 113295490 A CN113295490 A CN 113295490A
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- 206010020649 Hyperkeratosis Diseases 0.000 title claims abstract description 44
- 239000012188 paraffin wax Substances 0.000 title claims abstract description 30
- 241000425037 Toona sinensis Species 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims abstract description 28
- 235000011783 Cedrela sinensis Nutrition 0.000 title claims abstract description 23
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 145
- 239000001993 wax Substances 0.000 claims abstract description 17
- 239000000463 material Substances 0.000 claims abstract description 8
- 238000004043 dyeing Methods 0.000 claims abstract description 7
- 230000018044 dehydration Effects 0.000 claims abstract description 6
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 6
- 238000007598 dipping method Methods 0.000 claims abstract description 5
- 238000007789 sealing Methods 0.000 claims abstract description 5
- 238000001035 drying Methods 0.000 claims abstract description 3
- 235000019441 ethanol Nutrition 0.000 claims description 47
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 32
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 9
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000008096 xylene Substances 0.000 claims description 9
- 238000003892 spreading Methods 0.000 claims description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims description 3
- 102000002322 Egg Proteins Human genes 0.000 claims description 3
- 108010000912 Egg Proteins Proteins 0.000 claims description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 3
- 229960000583 acetic acid Drugs 0.000 claims description 3
- 235000014103 egg white Nutrition 0.000 claims description 3
- 210000000969 egg white Anatomy 0.000 claims description 3
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 claims description 3
- 239000008098 formaldehyde solution Substances 0.000 claims description 3
- 239000012362 glacial acetic acid Substances 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 230000007935 neutral effect Effects 0.000 claims description 3
- 230000001502 supplementing effect Effects 0.000 claims description 3
- 229950003937 tolonium Drugs 0.000 claims description 3
- HNONEKILPDHFOL-UHFFFAOYSA-M tolonium chloride Chemical compound [Cl-].C1=C(C)C(N)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 HNONEKILPDHFOL-UHFFFAOYSA-M 0.000 claims description 3
- 238000009966 trimming Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 3
- 230000008929 regeneration Effects 0.000 description 6
- 238000011069 regeneration method Methods 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 3
- 230000009261 transgenic effect Effects 0.000 description 3
- 244000025254 Cannabis sativa Species 0.000 description 2
- 235000012766 Cannabis sativa ssp. sativa var. sativa Nutrition 0.000 description 2
- 235000012765 Cannabis sativa ssp. sativa var. spontanea Nutrition 0.000 description 2
- 241000158728 Meliaceae Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 235000009120 camo Nutrition 0.000 description 2
- 235000005607 chanvre indien Nutrition 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 239000011487 hemp Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 240000001371 Chamaedaphne calyculata Species 0.000 description 1
- 235000013691 Chamaedaphne calyculata var. angustifolia Nutrition 0.000 description 1
- 235000013685 Chamaedaphne calyculata var. latifolia Nutrition 0.000 description 1
- 235000013687 Chamaedaphne calyculata var. nana Nutrition 0.000 description 1
- 235000011756 Vitis shuttleworthii Nutrition 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
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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
- G01N1/286—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
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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
- G01N1/286—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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- Health & Medical Sciences (AREA)
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- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
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Abstract
The invention discloses a method for preparing paraffin sections of cedrela sinensis callus, which comprises the steps of material fixation, alcohol dehydration, transparence, wax dipping, embedding, block repairing, block sticking and slicing, sheet unfolding and drying, dewaxing and transparence, rehydration and dyeing, sheet sealing, observation and the like. The invention innovates in the technical aspect according to the characteristics of tender and fragile cedrela sinensis callus, provides the paraffin section making method which is simple in operation process and particularly suitable for the cedrela sinensis callus, the obtained cedrela sinensis callus paraffin section is good in effect, simple in operation process and high in making efficiency, and meanwhile, the invention provides reference for making the paraffin sections of other plant callus. The invention has good application value.
Description
[ technical field ] A method for producing a semiconductor device
The invention relates to an efficient preparation method of paraffin sections, in particular to a preparation method of paraffin sections of cedrela sinensis callus.
[ background of the invention ]
Toona sinensis (A.Juss) Roem belongs to Meliaceae (Meliaceae) perennial deciduous trees, which are favored by light and grow well in fertile sandy soil with large day-night temperature difference. The cedrela sinensis not only can be used as an excellent fast-growing wood, but also is a delicious woody vegetable, and has extremely high nutritional and medicinal values. Under natural conditions, the propagation of the cedrela sinensis is mainly carried out by asexual propagation modes such as sowing, cutting seedling raising, tillering and the like, however, the propagation coefficients of the methods are low and the speed is slow. The traditional cultivation mode is difficult to meet the production requirement, and is not beneficial to the cultivation and popularization of improved varieties of the cedrela sinensis. Therefore, plant tissue culture becomes an important reproductive pathway.
In recent years, with the development of biotechnology, genetic engineering breeding opens up a new way for the improvement of Chinese toon varieties. The premise of establishing the high-efficiency transgenic system is the establishment of a high-efficiency regeneration system, and different regeneration systems of plants have direct influence on the transgenic efficiency. Therefore, the deep research on the regeneration system and the regeneration mode of the toona sinensis provides theoretical basis and technical support for the transgenic research of the toona sinensis. The cytological observation of the callus generated in the regeneration process of the Chinese toon by using a paraffin section technology is an important method for researching the regeneration mode of the Chinese toon.
At present, there are many techniques for making paraffin sections, and these techniques are different in terms of the characteristics of the tissues themselves, and the process flows of reagents, processing time and the like used in the process of making paraffin sections are also different. Chinese patent CN106769315B discloses a method for preparing paraffin sections of hemp callus, which comprises the steps of material taking and fixing, integral dyeing, dehydration, transparency, wax dipping and embedding, section cutting, sheet spreading and sticking, dewaxing, sheet sealing and the like, and the method can obtain the hemp callus paraffin sections with complete and clear tissue structures; chinese patent CN 102175482B discloses a thick leather leaf paraffin sectioning method, which comprises the steps of fixing, dehydrating, transparentizing, wax-permeating, embedding, sectioning, dewaxing, dyeing, and mounting, etc., wherein the method is used for selecting a fixing agent, determining a dehydration procedure, determining a transparentizing procedure, determining a wax-permeating procedure, and improving a dyeing and mounting method, and the method is also suitable for other leaves with different thicknesses and properties. However, no technique has been specifically applied to the slicing of the Toona sinensis tissue plants so far. The callus of the cedrela sinensis has the characteristics of tender and fragile, so that the conventional paraffin slicing method has poor effect, consumes time and is low in efficiency.
[ summary of the invention ]
The invention aims to solve the technical problem of overcoming the defects of the prior plant technology, and provides a paraffin section making method which is simple in operation process and particularly suitable for the callus of the cedrela sinensis according to the characteristics of tender and fragile cedrela sinensis callus and the like.
The invention provides a method for preparing paraffin sections of cedrela sinensis callus, which is characterized by comprising the following steps:
1) material fixation: at normal temperature, placing the cedrela sinensis callus in the improved FAA stationary liquid for fixing for 24 hours, and then exhausting air in a vacuum air extractor for 10-20 min to obtain fixed callus;
the improved FAA stationary liquid is a formaldehyde solution with the mass concentration of 37%: glacial acetic acid: ethanol solution with mass concentration of 70% is prepared by mixing ethanol solution with a volume ratio of 1: 1: 18 are mixed;
2) alcohol dehydration: dehydrating the fixed callus obtained in step 1) with 70% ethanol, 85% ethanol and 95% ethanol (containing eosin with a mass concentration of 1%) for 30min each stage; treating with anhydrous ethanol twice for 30min each time to obtain dehydrated callus;
3) and (3) transparency: transferring the dehydrated callus obtained in the step 2) into a bottle filled with absolute ethyl alcohol, and standing for 1 h; then sucking 1/4 volume of solution, adding chloroform with the same volume as the sucked solution, and standing for 1 h; then sucking 1/2 volume of solution, adding chloroform with the same volume as the sucked solution, and standing for 1 h; then sucking 3/4 volume of solution, adding chloroform with the same volume as the sucked solution, and standing for 1 h; finally, sucking all the solution away, supplementing chloroform with the same volume as the sucked solution, and standing for 1h to obtain transparent callus;
4) wax dipping: adding broken wax into the bottle in the step 3), adding broken wax with the mass gram of 4.5-5.5% of the volume milliliter of the solution in the bottle every 1 hour according to the principle of a small amount of multiple times until the added broken wax saturates the solution, and placing in a constant temperature box at 37 ℃ for 2 days;
5) embedding: the material was purified in chloroform: paraffin wax is mixed according to the volume ratio of 1: 3 for 30min at 52 ℃, then for 30min at 52 ℃ in pure paraffin, then for 30min at 48 ℃ in pure paraffin, and then for embedding;
6) trimming, sticking and slicing: transversely and longitudinally slicing the callus, wherein the thickness of the callus is 6-10 um, egg white glycerol is used as a sticky tablet, and distilled water is used for spreading the tablet;
7) unfolding and baking: spreading the slices at 37 ℃, and then drying in a constant temperature oven at 37 ℃ for 2-3 days;
8) dewaxing and transparency: dewaxing the xylene for 1h, and then replacing the xylene for transparency for 1 h;
9) rehydration and dyeing: treating with anhydrous ethanol, 95% ethanol, 85% ethanol, 70% ethanol, 50% ethanol, 30% ethanol, and distilled water for 30s per stage; 0.5% toluidine blue stain for 1 min;
10) mounting and observing: sequentially placing the glass slide into distilled water, 30% ethanol, 50% ethanol, 70% ethanol, 85% ethanol, 95% ethanol, anhydrous ethanol and xylene, treating for 30s per stage, and sealing with neutral gum; and placing the sealed slide in a ventilated place for 24 hours to obtain a permanent section.
The invention has the following beneficial effects: the paraffin section making technology which is particularly suitable for the callus of the Chinese toon is developed, the obtained paraffin section of the callus of the Chinese toon has good effect, simple operation flow and high making efficiency, and meanwhile, reference is provided for making the paraffin sections of other plant callus.
[ description of the drawings ]
FIG. 1 is a micrograph of a callus section of Toona sinensis according to an embodiment of the present invention.
[ detailed description ] embodiments
For a better understanding of the present invention, embodiments are described in detail below with reference to the accompanying drawings. It should be understood that this example is for illustrative purposes only and is not intended to limit the scope of the present invention.
The technical means adopted in the embodiment are conventional in the art, except for specific description.
Example 1
1) Material fixation: at normal temperature, placing the cedrela sinensis callus in the improved FAA stationary liquid for fixing for 24h, and then exhausting air in a vacuum air extractor for 10min to obtain fixed callus;
the improved FAA stationary liquid is prepared from a formaldehyde solution with a mass concentration of 37%, glacial acetic acid and an ethanol solution with a mass concentration of 70% in a volume ratio of 1: 1: 18 are mixed;
2) alcohol dehydration: dehydrating the fixed callus obtained in step 1) with 70% ethanol, 85% ethanol and 95% ethanol (containing eosin with a mass concentration of 1%) for 30min each stage; treating with anhydrous ethanol twice for 30min each time to obtain dehydrated callus;
3) and (3) transparency: transferring the dehydrated callus obtained in the step 2) into 4ml of absolute ethyl alcohol, and standing for 1 h; then sucking 1ml of solution by using a pipette gun, adding 1ml of chloroform, and standing for 1 h; then sucking away 2ml of solution, adding 2ml of chloroform, and standing for 1 h; then 3ml of the solution is sucked away, 3ml of chloroform is added, and the mixture is kept stand for 1 hour; finally, sucking away 4ml of solution, supplementing 4ml of chloroform, and standing for 1 h; replacing the solution in the bottle with pure chloroform to obtain transparent callus;
4) wax dipping: adding crushed wax into the bottle in the step 3), adding 0.18g of crushed wax every 1h according to the principle of a small amount of crushed wax for multiple times until the added crushed wax saturates the solution, and placing the solution in a constant temperature box at 37 ℃ for 2 days;
5) embedding: the material was purified in chloroform: paraffin wax is mixed according to the volume ratio of 1: 3 for 30min at 52 ℃, then for 30min at 52 ℃ in pure paraffin, then for 30min at 48 ℃ in pure paraffin, and then for embedding;
6) trimming, sticking and slicing: transversely and longitudinally slicing the callus by using an LEICA2150 slicer, wherein the thickness of the callus is 10um, egg white glycerol is used as a sticking tablet, and distilled water is used for spreading;
7) unfolding and baking: spreading the slices on a KD-H baking sheet machine at 37 ℃, and then baking the slices in a constant temperature oven at 37 ℃ for 2-3 days;
8) dewaxing and transparency: dewaxing the xylene for 1h, and then replacing the xylene for transparency for 1 h;
9) rehydration and dyeing: treating with anhydrous ethanol, 95% ethanol, 85% ethanol, 70% ethanol, 50% ethanol, 30% ethanol, and distilled water for 30s per stage; 0.5% toluidine blue stain for 1 min;
10) mounting and observing: sequentially placing the glass slide into distilled water, 30% ethanol, 50% ethanol, 70% ethanol, 85% ethanol, 95% ethanol, anhydrous ethanol and xylene, treating for 30s per stage, and sealing with neutral gum; and placing the sealed slide in a ventilated place for 24 hours to obtain a permanent section.
Example 2
This example is the same as example 1 with the difference that step 1) pumping time is 20min, step 4) adding 0.22g of crushed wax each time, step 6) slicing thickness is 6 um.
Example 3
This example is the same as example 1 or 2, except that the evacuation time in step 1) is 16min, 0.2g of crushed wax is added in each step in step 4), and the slice thickness in step 6) is 7 um.
Claims (1)
1. A method for preparing paraffin sections of cedrela sinensis callus is characterized by comprising the following steps:
1) material fixation: at normal temperature, placing the cedrela sinensis callus in the improved FAA stationary liquid for fixing for 24 hours, and then exhausting air in a vacuum air extractor for 10-20 min to obtain fixed callus;
the improved FAA stationary liquid is a formaldehyde solution with the mass concentration of 37%: glacial acetic acid: ethanol solution with mass concentration of 70% is prepared by mixing ethanol solution with a volume ratio of 1: 1: 18 are mixed;
2) alcohol dehydration: dehydrating the fixed callus obtained in step 1) with 70% ethanol, 85% ethanol and 95% ethanol (containing eosin with a mass concentration of 1%) for 30min each stage; treating with anhydrous ethanol twice for 30min each time to obtain dehydrated callus;
3) and (3) transparency: transferring the dehydrated callus obtained in the step 2) into a bottle filled with absolute ethyl alcohol, and standing for 1 h; then sucking 1/4 volume of solution, adding chloroform with the same volume as the sucked solution, and standing for 1 h; then sucking 1/2 volume of solution, adding chloroform with the same volume as the sucked solution, and standing for 1 h; then sucking 3/4 volume of solution, adding chloroform with the same volume as the sucked solution, and standing for 1 h; finally, sucking all the solution away, supplementing chloroform with the same volume as the sucked solution, and standing for 1h to obtain transparent callus;
4) wax dipping: adding broken wax into the bottle in the step 3), adding broken wax with the mass gram of 4.5-5.5% of the volume milliliter of the solution in the bottle every 1 hour according to the principle of a small amount of multiple times until the added broken wax saturates the solution, and placing in a constant temperature box at 37 ℃ for 2 days;
5) embedding: the material was purified in chloroform: paraffin wax is mixed according to the volume ratio of 1: 3 for 30min at 52 ℃, then for 30min at 52 ℃ in pure paraffin, then for 30min at 48 ℃ in pure paraffin, and then for embedding;
6) trimming, sticking and slicing: transversely and longitudinally slicing the callus, wherein the thickness of the callus is 6-10 um, egg white glycerol is used as a sticky tablet, and distilled water is used for spreading the tablet;
7) unfolding and baking: spreading the slices at 37 ℃, and then drying in a constant temperature oven at 37 ℃ for 2-3 days;
8) dewaxing and transparency: dewaxing the xylene for 1h, and then replacing the xylene for transparency for 1 h;
9) rehydration and dyeing: treating with anhydrous ethanol, 95% ethanol, 85% ethanol, 70% ethanol, 50% ethanol, 30% ethanol, and distilled water for 30s per stage; 0.5% toluidine blue stain for 1 min;
10) mounting and observing: sequentially placing the glass slide into distilled water, 30% ethanol, 50% ethanol, 70% ethanol, 85% ethanol, 95% ethanol, anhydrous ethanol and xylene, treating for 30s per stage, and sealing with neutral gum; and placing the sealed slide in a ventilated place for 24 hours to obtain a permanent section.
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| CN202110559019.XA CN113295490A (en) | 2021-05-21 | 2021-05-21 | Method for making paraffin section of cedrela sinensis callus |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114383906A (en) * | 2021-12-22 | 2022-04-22 | 中国农业科学院棉花研究所 | Preparation method of plant root system paraffin section |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106769315A (en) * | 2016-12-30 | 2017-05-31 | 中国农业科学院麻类研究所 | A kind of paraffin section preparation method of hemp callus |
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- 2021-05-21 CN CN202110559019.XA patent/CN113295490A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106769315A (en) * | 2016-12-30 | 2017-05-31 | 中国农业科学院麻类研究所 | A kind of paraffin section preparation method of hemp callus |
Non-Patent Citations (5)
| Title |
|---|
| 商园园: "红椿高效再生体系建立的研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》, no. 08, 15 August 2020 (2020-08-15), pages 12 - 13 * |
| 李巧峡: "《发育生物学实验指导》", 31 August 2019, 甘肃科学技术出版社, pages: 172 * |
| 王江: "广西香椿类型差异性研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》, no. 2, 15 February 2018 (2018-02-15), pages 11 - 13 * |
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| 赵丽萍: "《普通生物学实验》", 30 April 2018, 北京理工大学出版社, pages: 23 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114383906A (en) * | 2021-12-22 | 2022-04-22 | 中国农业科学院棉花研究所 | Preparation method of plant root system paraffin section |
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Application publication date: 20210824 |