CN108414318A - A kind of paraffin section production method of Rhododendron seeds - Google Patents
A kind of paraffin section production method of Rhododendron seeds Download PDFInfo
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- CN108414318A CN108414318A CN201810199156.5A CN201810199156A CN108414318A CN 108414318 A CN108414318 A CN 108414318A CN 201810199156 A CN201810199156 A CN 201810199156A CN 108414318 A CN108414318 A CN 108414318A
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- 241000208422 Rhododendron Species 0.000 title claims abstract description 47
- 239000012188 paraffin wax Substances 0.000 title claims abstract description 45
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 17
- 239000011521 glass Substances 0.000 claims abstract description 18
- 210000001672 ovary Anatomy 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 14
- 238000004043 dyeing Methods 0.000 claims abstract description 13
- 230000010152 pollination Effects 0.000 claims abstract description 7
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 claims description 53
- 239000001993 wax Substances 0.000 claims description 44
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 39
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 235000019441 ethanol Nutrition 0.000 claims description 18
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 16
- 230000018044 dehydration Effects 0.000 claims description 12
- 238000006297 dehydration reaction Methods 0.000 claims description 12
- 239000004902 Softening Agent Substances 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 8
- 235000011187 glycerol Nutrition 0.000 claims description 8
- 239000000975 dye Substances 0.000 claims description 6
- QUBBAXISAHIDNM-UHFFFAOYSA-N ethyldimethylbenzene Natural products CCC1=CC=CC(C)=C1C QUBBAXISAHIDNM-UHFFFAOYSA-N 0.000 claims description 6
- 239000004575 stone Substances 0.000 claims description 6
- 241001062009 Indigofera Species 0.000 claims description 5
- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 claims description 4
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Natural products C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 claims description 4
- 230000007935 neutral effect Effects 0.000 claims description 3
- 239000008096 xylene Substances 0.000 claims description 3
- 238000006424 Flood reaction Methods 0.000 claims description 2
- 238000004140 cleaning Methods 0.000 claims description 2
- 238000007598 dipping method Methods 0.000 claims description 2
- 238000005516 engineering process Methods 0.000 abstract description 3
- 230000007547 defect Effects 0.000 abstract 1
- 241000544061 Cuculus canorus Species 0.000 description 7
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 6
- 239000000463 material Substances 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 238000009396 hybridization Methods 0.000 description 3
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000009402 cross-breeding Methods 0.000 description 1
- 238000002845 discoloration Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 239000000203 mixture 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
-
- 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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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Fats And Perfumes (AREA)
Abstract
The present invention discloses a kind of paraffin section production method of Rhododendron seeds.This method is mainly characterized by, and the azalea ovary after pollination is taken just to be dyed after fixation, softening, is then dehydrated again, waxdip and embedding, slice, exhibition piece, bonding die, glass slide dry, dewax, mounting.It is clear that the method for the present invention can obtain dyeing, the complete Rhododendron seeds paraffin section of institutional framework, overcome existing paraffin section technology waxdip and embedding, slice, bonding die, open up just dye easily after piece, dewaxing cause a large amount of seed sheddings of process in dyeing to cause to be sliced in the incomplete defect of kernel texture.
Description
Technical field
The present invention relates to a kind of paraffin section technical field more particularly to a kind of paraffin section making sides of Rhododendron seeds
Method.
Background technology
Azalea is world's famous flower, and existing cuckoo can be effectively improved by carrying out distant hybridization breeding using its Wild ornamental resources
The ornamental value and its resistance of flower variety.But crossing barrier, especially after fertilization obstacle are that limitation azalea distant hybridization is successful
One major reason.Paraffin section technology can study hybrid embryo growth course from cellular level, to be educated for distant hybridization
Kind provides reliable basis.
Currently, the paraffin section making of indian azalea root, stem, leaf, fruit, seed is all made of following operational sequence：(1) it takes
Material, (2) are fixed, (3) softening, (4) dehydration, (5) are transparent, (6) waxdip and embedding, (7) slice, exhibition piece, bonding die, (8) dewaxing,
(9) it dyes, (10) are transparent, (11) mounting.Paraffin section is usually micron order thin slice, and the method is using the work for being first sliced poststaining
Sequence is operated, this operational sequence, for Rhododendron seeds, easily causes to cause in a large amount of seed sheddings of the process of dyeing
Keep kernel texture in slice imperfect.To solve this problem, need to research and develop a kind of paraffin section being suitable for Rhododendron seeds
Production method.
Invention content
Seed, which easily largely falls off, in paraffin section manufacturing process to solve existing Rhododendron seeds is difficult to obtain completely
The technical issues of seed tissue structure, a kind of dyeing of present invention offer is clear, the paraffin of the complete Rhododendron seeds of institutional framework
Microsection manufacture method.
A kind of paraffin section production method of Rhododendron seeds provided by the invention includes：Take the azalea ovary after pollination
It is soaked in FAA fixers and fixes, after taking-up is cleaned with clear water, be cut into along the axis length direction in the azalea ovary
Length is 0.2cm~0.5cm, and the stripping and slicing that width is 0.2cm~0.5cm and thickness is 0.2cm~0.5cm is put into softening agent
Impregnate softening, dyed and returned indigo plant after softening, then carry out successively dehydration and transparent, waxdip and embedding, slice, exhibition piece, bonding die,
Glass slide drying, dewaxing, mounting are the paraffin section that Rhododendron seeds are made；The dyeing is：The stripping and slicing softened is taken out, clearly
Washing 2~3 times, is soaked in haematoxylin dyeing liquid and dyes 3~7 days；Returning indigo plant is：Stained stripping and slicing is taken out, is put into clear water
It impregnates, change water to process is carried out during impregnating herein, it is non-discolouring until water and until stripping and slicing becomes navy blue.
Further, it is 7~30 days that the azalea ovary after the pollination, which is soaked in regular time in FAA fixers,.
Further, it is after cleaning 2~3 times with clear water after the taking-up is cleaned with clear water, described be put into softening agent is impregnated
Softening, the time for impregnating softening is 15~30 days, and the softening agent is 1 by volume ratio:1 70%v/v ethyl alcohol and glycerine is mixed
It closes.
Further, the dehydration with it is transparent be that stripping and slicing after returning basket is taken out, be first put into 50%v/v alcohol that impregnate 2 small
When, it places into 70%v/v alcohol overnight；Next day takes out stripping and slicing, is sequentially placed into 85%v/v alcohol, 95%v/v alcohol, anhydrous
Dehydration is impregnated in ethyl alcohol, impregnates dehydration 1 hour in each gradient solution；It is 2 to take out stripping and slicing from absolute ethyl alcohol and be first put into volume ratio:
It is impregnated 1 hour in 1 absolute ethyl alcohol and dimethylbenzene mixed liquor, it is 1 to place into new volume ratio:1 absolute ethyl alcohol and dimethylbenzene
It is impregnated 1 hour in mixed liquor, is then placed in pure dimethylbenzene and impregnates 1 hour, placed into new pure dimethylbenzene and impregnate 1 hour.
Further, the waxdip is that the stripping and slicing after being dehydrated and being transparent is put into container with embedding, and pure dimethylbenzene, which will be added, to be cut
Block floods, by pure dimethylbenzene:The mass ratio of rubble wax is 1:Rubble wax is added in 0.3 ratio, again by such as after the dissolving of rubble wax
This mode repeats that 2~3 rubble waxes are added, and is then put into container in 35 DEG C of baking ovens overnight；Oven temperature is adjusted to 42 by next day
DEG C, a rubble wax was added into container every 4 hours, adds rubble wax three times altogether, presses pure dimethylbenzene every time:The matter of rubble wax
Amount is than being 1:Rubble wax is added in 0.5 ratio, overnight at 42 DEG C later, 42 DEG C overnight after second day oven temperature is adjusted to 60
DEG C, first the solution in container is all outwelled, new atoleine is filled in container, interval is after 4 hours by the liquid in container
Body paraffin is all outwelled, and new atoleine is filled in container, and interval again all fell the atoleine in container after 4 hours
Fall, then fill new atoleine in container, later at 60 DEG C overnight, 60 DEG C overnight after second day, still maintain container
In the baking oven that temperature is 60 DEG C, first the atoleine in container is all outwelled, new atoleine is filled in container,
The atoleine in container is all outwelled after 4 hours, then fills new atoleine in container, interval is after 4 hours
Container is placed on progress paraffin embedding on paraffin wax embedding from taking-up in baking oven and obtains the wax stone containing stripping and slicing.
Further, the slice, exhibition piece, bonding die, glass slide drying are will to pass through waxdip to contain stripping and slicing with what embedding obtained
Wax stone be fixed on cycle type slicer, carrying out paraffin serial section after 8~10 microns of slice thickness of setting obtains wax disk(-sc), will
Wax disk(-sc), which is placed in exhibition piece in the water-bath that temperature is 42 DEG C, makes wax disk(-sc) be unfolded, and will into the water will after glass slide glycerine albumen smear
The glass slide for having glued wax disk(-sc) thereon, is finally put into 40 DEG C of insulating boxs and dries by the wax disk(-sc) adherency of expansion.
Further, it is described dewaxing, mounting be by drying after glass slide be first put into volume ratio be 1:1 absolute ethyl alcohol and two
Toluene mixture liquid is impregnated 15 minutes, places into pure xylene soak 25 minutes, is finally putting into new pure dimethylbenzene and is impregnated 35 points
Glass slide is taken out after clock, is the paraffin section that Rhododendron seeds are made with neutral gum mounting.
Compared with prior art, the beneficial effects of the invention are as follows：
There are multiple seeds in azalea cell, seed is tiny (be about 1500 μm, about 500 μm wide), between each seed
Independent, without close connection relation, therefore, Rhododendron seeds have caducous characteristics of organizational structure, according to existing cuckoo
The paraffin section production process of flower seed is dyed again after dehydration, waxdip and embedding, slice, bonding die, exhibition piece, dewaxing, that is, is contaminated
Color is to be sliced, after dewaxing, and it is all to be cut into micron order thin slice to be sliced usually, be sliced in addition after and is dewaxed, later again by it
Dyeing, dyeing impregnate a large amount of seeds of process easily fall off cause slice in kernel texture it is imperfect (its be sliced in seed not
Percentage of head rice reaches 60-80%), thus cannot get the Rhododendron seeds paraffin section of seed tissue structural integrity.The present invention is directed to
The above-mentioned biological property and Rhododendron seeds of Rhododendron seeds be a monoblock, frangible characteristics of organizational structure, will dye it is preposition,
Fix in FAA fixers, just dyed after softening agent softening, be then just dehydrated, waxdip and embedding, slice, bonding die,
Open up piece, drying, dewaxing, mounting process, it is ensured that the seed percentage of head rice in its slice reaches 100%, to easily can
It is clear to obtain dyeing, the complete Rhododendron seeds paraffin section of institutional framework, film-making is efficient, saves time and various materials
Material, produces unexpected technique effect, has developed a kind of paraffin section production method of suitable Rhododendron seeds, for Du
Cuckoo spends crossbreeding to provide reliable foundation.
Description of the drawings
Fig. 1 is the exploded chart that axis length direction of the azalea ovary in its ovary after pollination is cut into, and is marked in Fig. 1
1 indicates that the single cell of cuckoo, label 2 indicate that seed, label 3 indicate axis.
Fig. 2 is azalea ovary entirety sectional view, and label 1 indicates that the single cell of cuckoo, label 2 indicate seed in Fig. 2,
Label 3 indicates axis.
Fig. 3 is the single cell sectional view in azalea ovary, and label 1 indicates cuckoo single seed in Fig. 3.
Fig. 4 is the paraffin section figure of the Rhododendron seeds obtained using the method for the present invention, and label 1 indicates that cuckoo is single in Fig. 4
Grain seed.
Specific implementation mode
The present invention is described in detail with reference to embodiments, without specified otherwise is conventional method in embodiment.
A kind of paraffin section production method of Rhododendron seeds of the present invention, includes the following steps：
(1) it draws materials and fixed：The azalea ovary after pollination is taken to be soaked in FAA fixers fixed, according to ovary
Hardness fixes 7~30 days until the ovary interior tissue completely discoloration；The FAA fixers are by 70%v/v
Ethyl alcohol, formaldehyde and glacial acetic acid press 70%v/v ethyl alcohol:Formaldehyde:The volume ratio of glacial acetic acid is 18：1：1 ratio mixes.
(2) soften：The fixed azalea ovary will be passed through to take out, after washing 2~3 times with clear water, along the cuckoo
It is 0.2cm~0.5cm that length is cut into axis length direction in beggar room, and width is 0.2cm~0.5cm and thickness is 0.2cm
(since azalea ovary wall lignifying is higher, ovary tissues are harder, its ovary is carried out stripping and slicing, can make son for the stripping and slicing of~0.5cm
Room is easy softening), it is put into softening agent and impregnates 15~30 days until biopsy tissues soften；The softening agent is by volume ratio
1:1 70%v/v ethyl alcohol and glycerine mixes.
(3) it dyes：The material softened is taken out, clear water is washed 2~3 times, is soaked in haematoxylin dyeing liquid and dyes 3~7 days
Until stripping and slicing becomes kermesinus completely；The haematoxylin dyeing liquid is commercially available.
(4) indigo plant is returned：Stained stripping and slicing is taken out, is put into clear water and impregnates, change water to process is carried out during impregnating herein, until
Water is non-discolouring, until stripping and slicing is become navy blue by kermesinus.
(5) dehydration with it is transparent：The stripping and slicing after basket will be returned to take out, be first put into 50%v/v alcohol and impregnate 2 hours, place into
In 70%v/v alcohol overnight；Next day takes out stripping and slicing, is sequentially placed into 85%v/v alcohol 1 hour, is 95%v/v alcohol 1 hour, anhydrous
Ethyl alcohol impregnates dehydration in 1 hour；It is 2 to take out dewatered stripping and slicing from absolute ethyl alcohol later and be first put into volume ratio:1 absolute ethyl alcohol
It is impregnated 1 hour in dimethylbenzene mixed liquor, it is 1 to place into new volume ratio:It is impregnated in 1 absolute ethyl alcohol and dimethylbenzene mixed liquor
It 1 hour, is then placed in pure dimethylbenzene and impregnates 1 hour, place into new pure dimethylbenzene and impregnate 1 hour.
(6) waxdip and embedding：Stripping and slicing after being dehydrated and being transparent is put into container, pure dimethylbenzene is added and has just flooded stripping and slicing
Not yet, by pure dimethylbenzene (this waxdip and the pure dimethylbenzene being added in embedding step):The mass ratio of rubble wax is 1:0.3 ratio adds
Enter rubble wax, repeats that 3 rubble waxes are added by such mode again after the dissolving of rubble wax, then container is put into 35 DEG C of baking ovens
Overnight；Oven temperature is adjusted to 42 DEG C by next day, and a rubble wax was added into container every 4 hours, adds rubble wax three times altogether, often
It is secondary by pure dimethylbenzene (this waxdip and the pure dimethylbenzene that is added in embedding step):The mass ratio of rubble wax is 1:0.5 ratio
Be added rubble wax, later at 42 DEG C overnight, 42 DEG C overnight after second day oven temperature is adjusted to 60 DEG C, first take out container will hold
Solution in device is all outwelled, and is filled container in container after new atoleine (heating paraffin melt be atoleine)
60 DEG C of baking ovens are put into, interval takes out container and all outwells the atoleine in container after 4 hours, new liquid is filled in container
Container is put into 60 DEG C of baking ovens after body paraffin, interval further takes out container after 4 hours and all outwells the atoleine in container, then
Filled in container and container be put into 60 DEG C of baking ovens after new atoleine, overnight at 60 DEG C later, 60 DEG C overnight after
Second day, it is 60 DEG C to still maintain oven temperature, first takes out container and all outwells the atoleine in container, is fallen in container
Container is put into 60 DEG C of baking ovens after full new atoleine, container is taken out at interval after 4 hours the atoleine in container is whole
It outwells, then fills in container and container to be put into 60 DEG C of baking ovens after new atoleine, interval is after 4 hours by container from baking oven
Middle taking-up is placed on progress paraffin embedding on paraffin wax embedding and obtains the wax stone containing stripping and slicing.
(7) it is sliced, opens up piece, bonding die, glass slide drying：The wax stone containing stripping and slicing obtained with embedding by waxdip is fixed
Paraffin serial section is carried out on cycle type slicer, after the slice thickness of 8 microns of setting and obtains wax disk(-sc), and wax disk(-sc), which is placed in temperature, is
Piece is opened up in 42 DEG C of water-bath makes wax disk(-sc) be unfolded, and into the water will adhere to the wax disk(-sc) of expansion after glass slide glycerine albumen smear
Thereon, finally the glass slide for having glued wax disk(-sc) is put into 40 DEG C of insulating boxs and is dried to slide surface no moisture.The glycerine egg
It is in vain 1 by volume ratio:The 1 cleer and peaceful glycerine of Fresh Egg mixes.
(8) dewaxing, mounting：It is 1 that the glass slide dried in 40 DEG C of insulating boxs, which is first put into volume ratio,:1 absolute ethyl alcohol
With dimethylbenzene mixed liquid dipping 15 minutes, places into pure xylene soak 25 minutes, be finally putting into new pure dimethylbenzene and impregnate
Glass slide is taken out after 35 minutes, is the paraffin section that Rhododendron seeds are made with neutral gum mounting.
Claims (7)
1. a kind of paraffin section production method of Rhododendron seeds, it is characterised in that：The azalea ovary after pollination is taken to be soaked in
Fixed in FAA fixers, after taking-up is cleaned with clear water, being cut into length along the axis length direction in the azalea ovary is
0.2cm~0.5cm, the stripping and slicing that width is 0.2cm~0.5cm and thickness is 0.2cm~0.5cm, be put into softening agent impregnate it is soft
Change, dyed and returned indigo plant after softening, then carries out dehydration and transparent, waxdip and embedding, slice, exhibition piece, bonding die, glass slide successively
Drying, dewaxing, mounting are the paraffin section that Rhododendron seeds are made；The dyeing is：The stripping and slicing softened is taken out, clear water washes 2
It~3 times, is soaked in haematoxylin dyeing liquid and dyes 3~7 days；Returning indigo plant is：Stained stripping and slicing is taken out, is put into clear water and impregnates,
Change water to process is carried out during impregnating herein, it is non-discolouring until water and until stripping and slicing becomes navy blue.
2. the paraffin section production method of Rhododendron seeds according to claim 1, it is characterised in that：After the pollination
It is 7~30 days that azalea ovary, which is soaked in regular time in FAA fixers,.
3. the paraffin section production method of Rhododendron seeds according to claim 1, it is characterised in that：The taking-up is with clearly
That described be put into softening agent impregnates softening after clean 2~3 times with clear water after water cleaning, impregnate the time softened be 15~
30 days, the softening agent was 1 by volume ratio:1 70%v/v ethyl alcohol and glycerine mixes.
4. the paraffin section production method of Rhododendron seeds according to claim 1, it is characterised in that：It is described dehydration with thoroughly
Bright is that the stripping and slicing after returning basket is taken out, and is first put into 50%v/v alcohol and impregnates 2 hours, places into 70%v/v alcohol overnight；
Next day takes out stripping and slicing, is sequentially placed into 85%v/v alcohol, 95%v/v alcohol, absolute ethyl alcohol and impregnates dehydration, in each gradient solution
Impregnate dehydration 1 hour；It is 2 to take out stripping and slicing from absolute ethyl alcohol and be first put into volume ratio:In 1 absolute ethyl alcohol and dimethylbenzene mixed liquor
It impregnates 1 hour, it is 1 to place into new volume ratio:It impregnates 1 hour, is then placed in pure in 1 absolute ethyl alcohol and dimethylbenzene mixed liquor
It is impregnated 1 hour in dimethylbenzene, places into new pure dimethylbenzene and impregnate 1 hour.
5. the paraffin section production method of Rhododendron seeds according to claim 1, it is characterised in that：The waxdip and packet
It is that the stripping and slicing after being dehydrated and being transparent is put into container to bury, and pure dimethylbenzene is added and floods stripping and slicing, by pure dimethylbenzene:The matter of rubble wax
Amount is than being 1:Rubble wax is added in 0.3 ratio, repeats that 2~3 rubble waxes are added by such mode again after the dissolving of rubble wax, so
Container is put into 35 DEG C of baking ovens overnight afterwards；Oven temperature is adjusted to 42 DEG C by next day, is added into container every 4 hours primary broken
Paraffin adds rubble wax three times altogether, presses pure dimethylbenzene every time:The mass ratio of rubble wax is 1:Rubble wax is added in 0.5 ratio, it
Afterwards at 42 DEG C overnight, 42 DEG C overnight after second day oven temperature is adjusted to 60 DEG C, first the solution in container is all outwelled, it is past
New atoleine is filled in container, the atoleine in container is all outwelled after 4 hours, filled in container new by interval
Atoleine, interval again all outwelled the atoleine in container after 4 hours, then new atoleine is filled in container,
Overnight at 60 DEG C later, 60 DEG C overnight after second day, still maintain container is in 60 DEG C of baking oven in temperature, first will be in container
Atoleine all outwell, fill new atoleine in container, interval is whole by the atoleine in container after 4 hours
It outwells, then fills new atoleine in container, container is taken out from baking oven after 4 hours and is placed on paraffin wax embedding by interval
Upper progress paraffin embedding obtains the wax stone containing stripping and slicing.
6. the paraffin section production method of Rhododendron seeds according to claim 1, it is characterised in that：The slice, exhibition
Piece, bonding die, glass slide drying are that the wax stone containing stripping and slicing obtained with embedding by waxdip is fixed on cycle type slicer,
Paraffin serial section is carried out after the slice thickness of 8~10 microns of setting and obtains wax disk(-sc), and wax disk(-sc) is placed in the water-bath that temperature is 42 DEG C
Middle exhibition piece makes wax disk(-sc) be unfolded, and into the water will adhere to thereon the wax disk(-sc) of expansion after glass slide glycerine albumen smear, finally will
The glass slide for having glued wax disk(-sc) is put into 40 DEG C of insulating boxs and dries.
7. the paraffin section production method of Rhododendron seeds according to claim 1, it is characterised in that：The dewaxing, envelope
Piece be by drying after glass slide be first put into volume ratio be 1:1 absolute ethyl alcohol and dimethylbenzene mixed liquid dipping 15 minutes, then put
Enter pure xylene soak 25 minutes, is finally putting into after being impregnated 35 minutes in new pure dimethylbenzene and takes out glass slide, use neutral gum
Mounting is the paraffin section that Rhododendron seeds are made.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810195901 | 2018-03-09 | ||
| CN2018101959019 | 2018-03-09 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108414318A true CN108414318A (en) | 2018-08-17 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810199156.5A Pending CN108414318A (en) | 2018-03-09 | 2018-03-12 | A kind of paraffin section production method of Rhododendron seeds |
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| CN (1) | CN108414318A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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