CN102721583A - Method for producing film of leaf epidermis of stomatal pit of sweetscented oleander leaf - Google Patents
Method for producing film of leaf epidermis of stomatal pit of sweetscented oleander leaf Download PDFInfo
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- CN102721583A CN102721583A CN2012101784962A CN201210178496A CN102721583A CN 102721583 A CN102721583 A CN 102721583A CN 2012101784962 A CN2012101784962 A CN 2012101784962A CN 201210178496 A CN201210178496 A CN 201210178496A CN 102721583 A CN102721583 A CN 102721583A
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Abstract
The invention provides a method for producing a film of leaf epidermis of a stomatal pit of a sweetscented oleander leaf. The method comprises the steps of drawing materials and producing the film and further comprises the steps of the crosscutting film production of the sweetscented oleander leaf and the film production of the leaf epidermis at the bottom of the stomatal pit. In the step of the film production of the leaf epidermis at the bottom of the stomatal pit, mainly a micromanipulation method is utilized; a needle point with an inclined plane structure is used as dissection equipment; and after upper epidermis and mesophyll tissues on a superficial layer are picked and scrapped and when leaf veins and mesophyll tissues around the stomatal pit are picked and scrapped by the needle point, the mesophyll tissues attached onto the stomatal pit epidermis are scrapped by utilizing a cutting surface of the needle point. According to the invention, the film production of the leaf epidermis of the stomatal pit of the sweetscented oleander leaf is implemented by adopting the unarmed crosscutting film production of the leaf and the micromanipulation method; the method is simple to operate; the film production process is simplified; the film production time is shortened; and the shapes of leaf epidermis cells on the inner side walls and at the bottom of the stomatal pit of the sweetscented oleander leaf and the composition of a stomatal apparatus are observed.
Description
Technical field
[0001]
Technical field:
The invention belongs to biological technical field, be specifically related to a kind of flaking method of stomatal pit leaf epidermis of sweetscented oleander leaf.
Background technology:
Oleander (
Nerium indicumMill.) be a kind of frequently seen plants of Apocynaceae (Apocynaceae); In " botany ", " phytobiology " and all kinds of textbooks such as " phytotomies " and experimental teaching material, nearly all be to tell about as xerophytic typical Ye Lai with the blade construction of oleander.
The upper and lower epidermis of sweetscented oleander leaf all is a multiple layer epidermis, and its stomatal apparatus only is grown in the stomatal pit that sinks under the lower epidermis in the mesophyll tissue, can only see the opening of stomatal pit on the lower epidermis surface, can't observe stomatal apparatus and epidermis on every side thereof.Usually can only observe the longitudinal cutting structure of its stomatal pit and stomatal apparatus through the transverse section of sweetscented oleander leaf, cell shape and the stomatal apparatus that can't observe leaf epidermis in the stomatal pit constitute.Because this special construction of sweetscented oleander leaf stomatal pit makes all kinds of leaf epidermis flaking methods all can't obtain the leaf epidermis film-making of its stomatal pit, therefore the leaf epidermis film-making of oleander stomatal pit becomes a difficult problem on the leaf epidermis tabletting technology.
Summary of the invention:
For overcoming the shortcoming of above prior art, it is a kind of simple to operate that the present invention provides, and can observe the flaking method that interior leaf epidermal cell shape of oleander stomatal pit and stomatal apparatus constitute.
For achieving the above object, the present invention provides the flaking method of the stomatal pit leaf epidermis of a kind of oleander, and said method comprises the crosscut film-making of sweetscented oleander leaf and the leaf epidermis film-making of stomatal pit bottom for drawing materials and film-making.
The concrete steps of the crosscut film-making of sweetscented oleander leaf are following:
1) fresh sweetscented oleander leaf is placed in the visual field of anatomical lens, the part that earlier blade need not be observed cuts, and makes width of blade be about 1-1.5 cm;
2) along the direction not parallel or parallel, blade is carried out continuous crosscut with sharp cutter with the blade longitudinal axis;
3) after the section, will cut into slices with the tip tweezers and to transfer to successively on the microslide, process the water load, or pass through understain, film-making after washing.
The concrete steps of the leaf epidermis film-making of the stomatal pit bottom of sweetscented oleander leaf are following:
1) the fresh sweetscented oleander leaf of intercepting suitable length is placed in the visual field of anatomical lens with its epicuticle up, and transillumination is also suitably amplified the selection operation zone;
2) needle point with the 1mL syringe scratches epicuticle, and the mesophyll tissue with epicuticle and shallow-layer thereof chooses gently, scrapes off again; Drip in time dripping 1-2 on the operating surface simultaneously, or blade is put into the water flush away scrape residue and carry out moisturizing;
3) after epicuticle and part shallow-layer mesophyll tissue are removed, choose, scrape off the vein of all size and the mesophyll tissue around the stomatal pit with the needle point of 1mL syringe; Meanwhile, utilize the dip plane of needle point front end to scrape the mesophyll tissue on the air passing hole nest leaf epidermis lightly, scrape off lightly so that the spongy tissue that adheres on the stomatal pit leaf epidermis taken advantage of a situation;
4) after most mesophyll tissues in film-making zone are chosen, scrape off, this film-making zone is cut, transferred on the microslide, the water load is processed on the surface of its lower epidermis down, can observe and take a picture;
5) after transferring to leaf epidermis on the microslide, drip last water and dye liquor and carry out understain, make stomatal pit cell color on every side shallow slightly, and the dyeing of stomatal pit leaf epidermis is too not dark than the color of stomatal pit leaf epidermal cell; During dyeing, at the double dish that covers anti-dyeing liquor evaporation on the microslide; After the dyeing,, the water load is processed on the surface of lower epidermis down, can be observed and take a picture through rinsing.
In the leaf epidermis film-making process of sweetscented oleander leaf crosscut film-making and stomatal pit bottom, the dye liquor of understain all can be the 0.1% sarranine WS, 0.1% fast green dye liquor or other dye liquors.
Understain in the leaf epidermis film-making process of sweetscented oleander leaf crosscut film-making and stomatal pit bottom, the time of 0.1% sarranine WS understain all can be 5-10 minute when it dyeed, or according to Color, suitably shortened or prolong dyeing time
Understain in the leaf epidermis film-making process of sweetscented oleander leaf crosscut film-making and stomatal pit bottom, the time of 0.1% fast green dye liquor understain all can be 1-2 minute when it dyeed, or according to Color, suitably shortened or prolong dyeing time.
The present invention adopts the free-hand crosscut film-making and the micromanipulation method of leaf; Simplified the film-making program; Shortened the film-making time; Though the slice thick that the thickness of section is made than microtome some, through microscopical meticulous focusing, but increased the probability of observing stomatal pit sidewall leaf epidermal cell shape and stomatal apparatus formation when observing.Utilize this flaking method of the present invention; Even can not all mesophyll cells that adhere on the stomatal pit leaf epidermis thoroughly be removed; But the meticulous focusing that adjusts the telescope to one's eyes during through understain look and observation, leaf epidermal cell shape and the stomatal apparatus thereof that just can observe the stomatal pit bottom preferably constitute.
The present invention has solved the film-making difficult problem of oleander stomatal pit leaf epidermis through a kind of simple flaking method, has obtained the leaf epidermis film-making of oleander stomatal pit.Because oleander stomatal pit out-of-shape, bottom out-of-flatness, the epidermis staple length is and many, and stomatal apparatus all exists on the epidermis of stomatal pit bottom and sidewall, and stomatal apparatus is projecting inward to stomatal pit, and these factors make the spatial structure more complicated of oleander stomatal pit.When the leaf epidermal cell of oleander stomatal pit and stomatal apparatus are observed; To the leaf epidermal cell observation of stomatal pit opening, stomatal pit profile, stomatal pit bottom be combined, could produce a solid and comprehensive understanding to the leaf epidermal cell shape of oleander stomatal pit and the structure of stomatal apparatus like this.
Description of drawings:
Fig. 1: the lower epidermis of sweetscented oleander leaf, show stomatal pit opening (Fluirescence observation);
Fig. 2: the amplification of a part of stomatal pit opening (dyeing);
Fig. 3: the amplification of a stomatal pit opening (dyeing), photo is through optical information processing and enlarged drawing;
Fig. 4: the longitudinal section of a stomatal pit;
Fig. 5: the longitudinal section of a stomatal pit (Fluirescence observation);
Fig. 6: the longitudinal section of a stomatal pit (Fluirescence observation);
Fig. 7: the longitudinal section of a stomatal pit (Fluirescence observation);
Fig. 8: the longitudinal section (Fluirescence observation) of the stomatal pit of two opening merging;
Fig. 9: be arranged in two stomatal pits (Fluirescence observation) mesophyll tissue, that do not link to each other with epidermis;
Figure 10 a: stomatal pit that is arranged in mesophyll tissue;
Figure 11: the leaf epidermis film-making of oleander stomatal pit bottom, show a leaf epidermis (being unstained) bottom the complete stomatal pit
Figure 12: the amplification of last figure, show the part leaf epidermis and the stomatal apparatus of stomatal pit bottom;
Figure 13: the leaf epidermis of a stomatal pit bottom and the enlarged drawing of stomatal apparatus;
Figure 14: the leaf epidermis and the stomatal apparatus (dyeing) of a stomatal pit bottom;
Figure 15: the amplification of the part leaf epidermis of Figure 14 stomatal pit bottom;
Figure 16: the part leaf epidermis of a stomatal pit bottom and the amplification (dyeing) of stomatal apparatus;
Figure 17: the leaf epidermal cell and the stomatal apparatus (Fluirescence observation) of a stomatal pit bottom;
The stomatal pit of Figure 18: Figure 10 amplifies;
Figure 19: the stomatal apparatus on the stomatal pit sidewall;
Figure 20: the stomatal apparatus on the stomatal pit sidewall;
Figure 21: the stomatal apparatus on the stomatal pit sidewall (Fluirescence observation);
The stomatal apparatus on the stomatal pit sidewall is shown in the amplification of Figure 22: Figure 19;
Figure 23: the amplification of the stomatal apparatus on the stomatal pit sidewall;
The stomatal apparatus on the stomatal pit sidewall is shown in the amplification of Figure 24: Figure 21.
Embodiment
Below in conjunction with embodiment flaking method of the present invention is done further to specify.
Embodiment 1:
A kind of flaking method of stomatal pit leaf epidermis of sweetscented oleander leaf, said method is for drawing materials and film-making, and blade picks up from the oleander of planting in Luoyang City, the Henan Province garden, and its flaking method comprises the crosscut film-making of sweetscented oleander leaf and the leaf epidermis film-making of stomatal pit bottom.
1. the crosscut film-making of sweetscented oleander leaf
1) fresh sweetscented oleander leaf is placed in the visual field of anatomical lens, the part that earlier blade need not be observed cuts, and makes width of blade be about 1 cm;
2) along the direction parallel with the blade longitudinal axis, with sharp cutter blade is carried out continuous crosscut, transverse section is thinner as far as possible;
3) after the section, will cut into slices with the tip tweezers and to transfer to successively on the microslide, process water load
.
2. the film-making of the stomatal pit of sweetscented oleander leaf bottom leaf epidermis
1) the fresh sweetscented oleander leaf of intercepting suitable length is placed in the visual field of anatomical lens with its epicuticle up, with transillumination and suitably amplification, and the selection operation zone, left hand is pushed down the oleander blade, and the right hand begins to operate with the 1mL syringe;
2) needle point with the 1mL syringe scratches epicuticle, and the mesophyll tissue of epicuticle and shallow-layer thereof is chosen gently, scrapes off, and does not injure the leaf epidermal cell of stomatal pit as far as possible; Leaf epidermis dehydration shrinkage in order to prevent to operate is dripped in time dripping 2 on the operating surface simultaneously;
3) after epicuticle and part shallow-layer mesophyll tissue are removed, choose, scrape off the vein of all size and the mesophyll tissue around the stomatal pit lightly with the needle point of 1mL syringe; Meanwhile, utilize the dip plane of needle point front end to scrape the mesophyll tissue on the air passing hole nest leaf epidermis lightly, scrape off lightly so that the spongy tissue that adheres on the stomatal pit leaf epidermis taken advantage of a situation;
4) after most mesophyll tissues in film-making zone are chosen, scrape off, this film-making zone is cut, transferred on the microslide, the water load is processed on the surface of its lower epidermis down, can observe and take a picture;
5) after transferring to leaf epidermis on the microslide, drip the last water and the 0.1% sarranine WS and carried out understain 6 minutes, make stomatal pit cell color on every side shallow slightly, and the dyeing of stomatal pit leaf epidermis is too not dark than the color of stomatal pit leaf epidermal cell; During dyeing, at the double dish that covers anti-dyeing liquor evaporation on the microslide; After the dyeing,, the water load is processed on the surface of lower epidermis down, can be observed and take a picture through rinsing.
Embodiment 2:
A kind of flaking method of stomatal pit leaf epidermis of sweetscented oleander leaf, said method is for drawing materials and film-making, and blade picks up from the oleander of planting in Luoyang City, the Henan Province garden, and its flaking method comprises the crosscut film-making of sweetscented oleander leaf and the leaf epidermis film-making of stomatal pit bottom.
1. the crosscut film-making of sweetscented oleander leaf
1) fresh sweetscented oleander leaf is placed in the visual field of anatomical lens, the part that earlier blade need not be observed cuts, and makes width of blade be about 1.2 cm;
2) along the direction vertical with the blade longitudinal axis, with sharp cutter blade is carried out continuous crosscut, transverse section is thinner as far as possible;
3) after the section, will cut into slices with the tip tweezers and to transfer to successively on the microslide, through 0.1% sarranine dye liquor understain 5 minutes, film-making after washing then.
2. the film-making of the stomatal pit of sweetscented oleander leaf bottom leaf epidermis
1) the fresh sweetscented oleander leaf of intercepting suitable length is placed in the visual field of anatomical lens with its epicuticle up, with transillumination and suitably amplification, and the selection operation zone, left hand is pushed down the oleander blade, and the right hand begins to operate with the 1mL syringe;
2) needle point with the 1mL syringe scratches epicuticle, and the mesophyll tissue of epicuticle and shallow-layer thereof is chosen gently, scrapes off, and does not injure the leaf epidermal cell of stomatal pit as far as possible; Leaf epidermis dehydration shrinkage in order to prevent to operate is dripped in time dripping 1 on the operating surface simultaneously;
3) after epicuticle and part shallow-layer mesophyll tissue are removed, choose, scrape off the vein of all size and the mesophyll tissue around the stomatal pit lightly with the needle point of 1mL syringe; Meanwhile, utilize the dip plane of needle point front end to scrape the mesophyll tissue on the air passing hole nest leaf epidermis lightly, scrape off lightly so that the spongy tissue that adheres on the stomatal pit leaf epidermis taken advantage of a situation;
4) after most mesophyll tissues in film-making zone are chosen, scrape off, this film-making zone is cut, transferred on the microslide, the water load is processed on the surface of its lower epidermis down, can observe and take a picture;
5) after transferring to leaf epidermis on the microslide, drip the last water and the 0.1% sarranine WS and carried out understain 10 minutes, make stomatal pit cell color on every side shallow slightly, and the dyeing of stomatal pit leaf epidermis is too not dark than the color of stomatal pit leaf epidermal cell; During dyeing, at the double dish that covers anti-dyeing liquor evaporation on the microslide; After the dyeing,, the water load is processed on the surface of lower epidermis down, can be observed and take a picture through rinsing.
Embodiment 3:
A kind of flaking method of stomatal pit leaf epidermis of sweetscented oleander leaf, said method is for drawing materials and film-making, and blade picks up from the oleander of planting in Luoyang City, the Henan Province garden, and its flaking method comprises the crosscut film-making of sweetscented oleander leaf and the leaf epidermis film-making of stomatal pit bottom.
1. the crosscut film-making of sweetscented oleander leaf
1) fresh sweetscented oleander leaf is placed in the visual field of anatomical lens, the part that earlier blade need not be observed cuts, and makes width of blade be about 1.4 cm;
2) along with the uneven direction of the blade longitudinal axis, with sharp cutter blade is carried out continuous crosscut, transverse section is thinner as far as possible;
3) after the section, will cut into slices with the tip tweezers and to transfer to successively on the microslide, through 0.1% sarranine dye liquor understain 10 minutes, film-making after washing then.
2. the film-making of the stomatal pit of sweetscented oleander leaf bottom leaf epidermis
1) the fresh sweetscented oleander leaf of intercepting suitable length is placed in the visual field of anatomical lens with its epicuticle up, with transillumination and suitably amplification, and the selection operation zone, left hand is pushed down the oleander blade, and the right hand begins to operate with the 1mL syringe;
2) needle point with the 1mL syringe scratches epicuticle, and the mesophyll tissue of epicuticle and shallow-layer thereof is chosen gently, scrapes off, and does not injure the leaf epidermal cell of stomatal pit as far as possible; Leaf epidermis dehydration shrinkage is put into the water flush away with blade simultaneously and is scraped residue and carry out moisturizing in order to prevent to operate;
3) after epicuticle and part shallow-layer mesophyll tissue are removed, choose, scrape off the vein of all size and the mesophyll tissue around the stomatal pit lightly with the needle point of 1mL syringe; Meanwhile, utilize the dip plane of needle point front end to scrape the mesophyll tissue on the air passing hole nest leaf epidermis lightly, scrape off lightly so that the spongy tissue that adheres on the stomatal pit leaf epidermis taken advantage of a situation;
4) after most mesophyll tissues in film-making zone are chosen, scrape off, this film-making zone is cut, transferred on the microslide, the water load is processed on the surface of its lower epidermis down, can observe and take a picture;
5) after transferring to leaf epidermis on the microslide, drip about 2 minutes of the time that last water and 0.1% fast green dye liquor carry out understain, make stomatal pit cell color on every side shallow slightly, and the dyeing of stomatal pit leaf epidermis is too not dark than the color of stomatal pit leaf epidermal cell; During dyeing, at the double dish that covers anti-dyeing liquor evaporation on the microslide; After the dyeing,, the water load is processed on the surface of lower epidermis down, can be observed and take a picture through rinsing.
Embodiment 4:
A kind of flaking method of stomatal pit leaf epidermis of sweetscented oleander leaf, said method is for drawing materials and film-making, and blade picks up from the oleander of planting in Luoyang City, the Henan Province garden, and its flaking method comprises the crosscut film-making of sweetscented oleander leaf and the leaf epidermis film-making of stomatal pit bottom.
1. the crosscut film-making of sweetscented oleander leaf
1) fresh sweetscented oleander leaf is placed in the visual field of anatomical lens, the part that earlier blade need not be observed cuts, and makes width of blade be about 1.5 cm;
2) along the direction parallel with the blade longitudinal axis, with sharp cutter blade is carried out continuous crosscut, transverse section is thinner as far as possible;
3) after the section, will cut into slices with the tip tweezers and to transfer to successively on the microslide, through 0.1% fast green dye liquor understain 2 minutes, film-making after washing then.
2. the film-making of the stomatal pit of sweetscented oleander leaf bottom leaf epidermis
1) the fresh sweetscented oleander leaf of intercepting suitable length is placed in the visual field of anatomical lens with its epicuticle up, with transillumination and suitably amplification, and the selection operation zone, left hand is pushed down the oleander blade, and the right hand begins to operate with syringe;
2) needle point with syringe scratches epicuticle; The mesophyll tissue of epicuticle and shallow-layer thereof is chosen gently, scrapes off; Do not injure the leaf epidermal cell of stomatal pit as far as possible, leaf epidermis dehydration shrinkage is put into the water flush away with blade simultaneously and is scraped residue and carry out moisturizing in order to prevent to operate;
3) after epicuticle and part shallow-layer mesophyll tissue are removed, choose, scrape off the vein of all size and the mesophyll tissue around the stomatal pit lightly with the needle point of syringe; Meanwhile, utilize the dip plane of needle point front end to scrape the mesophyll tissue on the air passing hole nest leaf epidermis lightly, scrape off lightly so that the spongy tissue that adheres on the stomatal pit leaf epidermis taken advantage of a situation;
4) after most mesophyll tissues in film-making zone are chosen, scrape off, this film-making zone is cut, transferred on the microslide, the water load is processed on the surface of its lower epidermis down, can observe and take a picture;
5) after transferring to leaf epidermis on the microslide, drip about 1 minute of the time that last water and 0.1% fast green dye liquor carry out understain, make stomatal pit cell color on every side shallow slightly, and the dyeing of stomatal pit leaf epidermis is too not dark than the color of stomatal pit leaf epidermal cell; During dyeing, at the double dish that covers anti-dyeing liquor evaporation on the microslide; After the dyeing,, the water load is processed on the surface of lower epidermis down, can be observed and take a picture through rinsing.
The optical information processing method of photograph and part digital photograph
When film-making is observed, use fluorescent microscope in the normal optical and the fluorescence visual field, to observe.Adjust the telescope to one's eyes or anatomical lens under observations when taking a picture, digital camera is set to " microshot " pattern earlier, and closes flashlamp, uses digital camera directly to take pictures facing to the eyepiece of microscope or anatomical lens then.After choosing the observations that to take pictures, after the enlarging function that can also utilize digital camera to carry carries out clear amplification to image, take pictures again.For making structure observation clearer, indivedual digital photographs have adopted the optical information analytic method to handle, but most digital photograph is unprocessed.
Result and analysis
1. the observation of oleander stomatal pit
The opening of oleander stomatal pit on lower epidermis irregular (Fig. 1); The opening of some stomatal pits is about triangular shape circular or oval (Fig. 2); In the opening and stomatal pit of stomatal pit, some elongated epidermal hairs that in the stomatal pit chamber, stretched out by stomatal pit epidermal cell outer wall (Fig. 3-4) are arranged.
See from the longitudinal section of oleander stomatal pit, the out-of-shape in stomatal pit chamber, bottom out-of-flatness (Fig. 4-7), the opening of visible stomatal pit merges, is grown in together (Fig. 1,8) sometimes.On the longitudinal section of stomatal pit, have some stomatal pits do not see and lower epidermis between the duct (Fig. 9-10) got in touch, the opening that stomatal pit is described is than stomatal pit chamber little or short (Fig. 5,7,9) generally.
A rip cutting stomatal pit that is arranged in mesophyll tissue is observed, and the bottom of visible stomatal pit links to each other with spongy tissue, and its upper end is palisade tissue (Figure 10).
2. the observation of oleander stomatal pit leaf epidermis and stomatal apparatus
After choosing, scrape off the epicuticle and mesophyll tissue of oleander, not dyed or film-making behind the understain look, leaf epidermal cell shape and the stomatal apparatus that can observe oleander stomatal pit bottom constitute (Figure 11-16).
Fluirescence observation is carried out in leaf epidermis film-making to oleander stomatal pit bottom, can be observed circular stomatal apparatus profile and epidermal hair on the stomatal pit leaf epidermis at supracutaneous opening, and two guard cells' form can be distinguished (Figure 17).Stomatal pit to Figure 10 amplifies observation; It is thus clear that the leaf epidermis in the stomatal pit is made up of cell monolayer; On the epidermis of stomatal pit bottom; The cell that constitutes stomatal apparatus significantly protrudes in the stomatal pit chamber, has elongated epidermal hair to stretch out from the outer wall of stomatal pit leaf epidermal cell around it, extend into (Figure 18) in the stomatal pit chamber.
On the transverse section of leaf, can be observed on the longitudinal cutting structure of some stomatal pits, the surface that part leaf epidermis and stomatal apparatus have occurred is seen, and this is leaf epidermis and stomatal apparatus (Figure 19-24) on the stomatal pit sidewall.See on the leaf epidermis of stomatal pit bottom and sidewall, stomatal apparatus (Figure 18-21) is arranged all from observations.
We can draw through above embodiment; The present invention adopts the free-hand crosscut film-making and the micromanipulation method of leaf; Simplified the film-making program; Shortened the film-making time, leaf epidermal cell shape and the stomatal apparatus observed in the oleander stomatal pit constitute, thereby make people produce a solid and comprehensive understanding to the leaf epidermal cell shape of oleander stomatal pit and the structure of stomatal apparatus.
Claims (6)
1. the flaking method of the stomatal pit leaf epidermis of a sweetscented oleander leaf is characterized in that: said method is for drawing materials and film-making, and flaking method comprises the crosscut film-making of sweetscented oleander leaf and the leaf epidermis film-making of stomatal pit bottom.
2. flaking method according to claim 1 is characterized in that the concrete steps of crosscut film-making of bamboo peach leaf are following:
1) fresh sweetscented oleander leaf is placed in the visual field of anatomical lens, the part that earlier blade need not be observed cuts, and makes width of blade be about 1-1.5 cm;
2) along the direction not parallel or parallel, blade is carried out continuous crosscut with blade with the blade longitudinal axis;
3) after the section, will cut into slices with the tip tweezers and to transfer to successively on the microslide, process the water load, or, wash film-making afterwards through understain.
3. flaking method according to claim 1 is characterized in that the concrete steps of leaf epidermis film-making of stomatal pit bottom of sweetscented oleander leaf are following:
1) the fresh sweetscented oleander leaf of intercepting suitable length is placed in the visual field of anatomical lens with its epicuticle up, and transillumination is also suitably amplified the selection operation zone;
2) epicuticle is scratched, the mesophyll tissue with epicuticle and shallow-layer thereof chooses gently, scrapes off again; Drip in time dripping 1-2 on the operating surface simultaneously, or blade is put into the water flush away scrape residue and carry out moisturizing;
3) after epicuticle and part shallow-layer mesophyll tissue are removed, choose, scrape off the vein of all size and the mesophyll tissue around the stomatal pit; Meanwhile, utilize the dip plane of needle point front end to scrape the mesophyll tissue on the air passing hole nest leaf epidermis lightly, scrape off lightly so that the spongy tissue that adheres on the stomatal pit leaf epidermis taken advantage of a situation;
4) after most mesophyll tissues in film-making zone are chosen, scrape off, this film-making zone is cut, transferred on the microslide, the water load is processed on the surface of its lower epidermis down, can observe and take a picture;
5) after transferring to leaf epidermis on the microslide, drip last water and dye liquor and carry out understain; After the dyeing,, the water load is processed on the surface of lower epidermis down, can be observed and take a picture through rinsing.
4. according to claim 1,2 or 3 described flaking methods, it is characterized in that the dye liquor of understain in the leaf epidermis film-making process of sweetscented oleander leaf crosscut film-making and stomatal pit bottom all can be the 0.1% sarranine WS, 0.1% fast green dye liquor or other dye liquors.
5. according to claim 1,2 or 3 described flaking methods, it is characterized in that understain in the leaf epidermis film-making process of sweetscented oleander leaf crosscut film-making and stomatal pit bottom, the time of 0.1% sarranine WS understain all can be 5-10 minute when it dyeed.
6. according to claim 1,2 or 3 described flaking methods, it is characterized in that understain in the leaf epidermis film-making process of sweetscented oleander leaf crosscut film-making and stomatal pit bottom, the time of 0.1% fast green dye liquor understain all can be 1-2 minute when it dyeed.
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| CN104596815A (en) * | 2014-12-17 | 2015-05-06 | 西南林业大学 | Bamboo sliding section staining production method |
| CN105547740A (en) * | 2015-12-10 | 2016-05-04 | 大连大学 | Method for acquiring epidermis of plant leaf |
| CN107179268A (en) * | 2017-05-26 | 2017-09-19 | 安徽科技学院 | A kind of estimating and measuring method of Leaf-Blade of Poaceae epidermis stomatal frequency |
| CN109085046A (en) * | 2018-10-25 | 2018-12-25 | 南京林业大学 | A kind of preparation method of leaf of bamboo internal anatomy 3 dimensional drawing |
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| CN105547740A (en) * | 2015-12-10 | 2016-05-04 | 大连大学 | Method for acquiring epidermis of plant leaf |
| CN107179268A (en) * | 2017-05-26 | 2017-09-19 | 安徽科技学院 | A kind of estimating and measuring method of Leaf-Blade of Poaceae epidermis stomatal frequency |
| CN109085046A (en) * | 2018-10-25 | 2018-12-25 | 南京林业大学 | A kind of preparation method of leaf of bamboo internal anatomy 3 dimensional drawing |
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