CN109425521A - A kind of preparation method of phytolite Dispersed Cuticles - Google Patents
A kind of preparation method of phytolite Dispersed Cuticles Download PDFInfo
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- CN109425521A CN109425521A CN201710748834.4A CN201710748834A CN109425521A CN 109425521 A CN109425521 A CN 109425521A CN 201710748834 A CN201710748834 A CN 201710748834A CN 109425521 A CN109425521 A CN 109425521A
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- sample
- cuticles
- dispersed
- cuticula
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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
- G01N1/2806—Means for preparing replicas of specimens, e.g. for microscopal analysis
Abstract
The present invention provides a kind of preparation methods of phytolite Dispersed Cuticles, include: to Dispersed Cuticles sample pre-treatments: by choosing cuticula sample and removing the water-solubility impurity in sample, carbonate components and siliceous ingredient, obtaining pre-treatment Dispersed Cuticles sample；Then Dispersed Cuticles oxidation processes are obtained into the Dispersed Cuticles sample of oxidation processes；The upper and lower epidermis separation of Dispersed Cuticles is finally carried out, cuticula is obtained.The present invention is realized the quick preparation of phytolite Dispersed Cuticles using infusion method, effectively shortens experimental period, improve the quality for the cuticula prepared in the preparation process of phytolite Dispersed Cuticles.
Description
Technical field
The invention belongs to paleobotany technical fields, are related to a kind of preparation method of phytolite Dispersed Cuticles.
Background technique
Cuticula is present in the outer wall of plant surface chrotoplast, is one layer of fatty protective layer.Previous fossil plant angle
Matter layer material is taken from mostly on compression sample, for cuticula in situ.Those have disengaged from plant parent or plant in stone
It is carried over long distances before changing and ruptures fragments of stratum corneum of the distributed and saved in sedimentary type formations, referred to as Dispersed Cuticles.Dispersion angle
Matter layer feature can be used for plant identification monoid or kind, also can be used as one of Strata Comparison, ancient ecological Studies important means.
But Dispersed Cuticles are often very fragile by very long geological process, it is difficult to prepare, to seriously constrain correlative study
Carry out.
The experimental method of Ye Meina short summary fossil cuticle processing, mainly includes Shu Shi solution (Schulze
Solution method, He Defa (Heard Method, i.e. liquor natrii hypochloritis are macerated method) and hydrofluoric acid) are macerated and is macerated method.Wherein,
Shu Shi solution is macerated method and He Defa uses the strong oxidizers Stratum Corneum such as Shu Shi solution or liquor natrii hypochloritis to carry out oxygen first
Change processing, followed by lye (Ammonia, sodium hydroxide solution or potassium hydroxide solution) by the mesophyll group after oxidation
Precipitation is knitted, to achieve the purpose that prepared by cuticula.Above two method is in situ in cuticula research using relatively broad.But
Since Dispersed Cuticles type is abundant, different type cuticula is to Shu Shi solution, liquor natrii hypochloritis, Ammonia (ammonia
Water) etc. reaction sensitivity, the reaction time be not quite similar, when easily cuticula being caused to encounter mentioned reagent occur shrinkage, volume
Bent or rupture.Hydrofluoric acid be macerated rule be will the glue through adhering on above two method treated cuticula be siliceous and fluorination
The dissolution of the substances such as silicon, keeps cuticula more pure, but the cuticula through aoxidizing is extremely fragile, after hydrofluoric acid solution is handled easily
Damage.In addition, above-mentioned three kinds of methods experiment periods are longer, this also constrains its reality in Dispersed Cuticles research and answers
With.
Summary of the invention
In order to overcome the disadvantages of prior art experimental period is long, experiment effect is undesirable, the present invention provides a kind of fossils
The preparation method of plant Dispersed Cuticles can be quickly obtained high quality cuticula sample, for epidermal shape identification, stratum pair
Material foundation is provided than, ancient ecological Studies.
The present invention realizes by the following method:
A kind of preparation method of phytolite Dispersed Cuticles, comprising:
Step A: it chooses cuticula sample and removes the water-solubility impurity in sample, carbonate components and siliceous ingredient, obtain
To pre-treatment Dispersed Cuticles sample；
Step B: the pre-treatment Dispersed Cuticles sample that step A is obtained being placed in Shu Shi solution and is impregnated, until leaf film browning
Reaction is terminated when color is translucent or transparent, is washed to neutrality, is obtained the Dispersed Cuticles sample of oxidation processes；
Step C: after the Dispersed Cuticles samples with water by oxidation processes is soaked for a period of time, hydroxide is added
Ammonium salt solution, until terminate reaction after remaining mesophyll tissue and vascular bundle becomes brown floccule or filiform, be washed to neutrality,
It takes out the Dispersed Cuticles sample and removes upper and lower epidermis, obtain cuticula；
Step D: removing the impurity on resulting cuticula sample and clean cuticula, is subsequently placed in water and impregnates after
Continuous film-making observation.
According to the preferred embodiment of the present invention, the Shu Shi solution includes being saturated potassium chlorate solution and fuming nitric aicd, excellent
The volume ratio of selection of land, the saturation potassium chlorate solution and fuming nitric aicd is 1:(2-3).Shu Shi solution used in the present invention ensures
Stratum Corneum does not destroy during the treatment.
In the present invention, in step A, water-solubility impurity is removed with distilled water；And/or with acid solution remove carbonate at
Divide, preferably hydrochloric acid；And/or siliceous ingredient is removed with hydrofluoric acid solution.It is to eliminate in country rock with the purpose of HCl treatment
Carbonate mineral prevents the calcareous reaction in hydrofluoric acid maceration from generating calcirm-fluoride and is attached to cutin layer surface and causes to prepare
Cuticula can not clearly observe its structure by impurity covering.
According to the preferred embodiment of the present invention, in step A, Dispersed Cuticles sample pre-treatments include:
Step A-1, the cuticula sample of selection is placed in sequence in the container with multiple sample cells, adds distillation
Water impregnates the cuticula sample 40-60min, then removes the distilled water in sample cell；
Step A-2, hydrochloric acid solution is added and impregnates 3-4h, then hydrochloric acid is removed and washes the cuticula sample into
Property；
Step A-3, hydrofluoric acid solution is added to impregnate 12-24 hours, until country rock becomes cotton-shaped or powdered completely, with distillation
Water cleans cotton-shaped or powdered country rock repeatedly to neutrality.
In the present invention, in the step A-1 distilled water immersion preferably carried out at 40-60 DEG C and/or step A-2 in salt
The concentration of acid be 35-40% and/or step A-3 described in the volumetric concentration of hydrofluoric acid be 40-45%, preferably hydrofluoric acid dips
At 40-60 DEG C, preferably carried out at 40 DEG C.
According to the preferred embodiment of the present invention, preferred higher than the operation carried out under room temperature in the step A-1
It is carried out in a manner of heating water bath.
According to the preferred embodiment of the present invention, hydrofluoric acid is preferably to glassware corrosion, the container in order to prevent
Makrolon material is made.The container with multiple sample cells that the present invention uses, being operated simultaneously using the time difference can be improved
Efficiency.
In the application in Dispersed Cuticles pretreatment process, in the step of acid solution removing carbonate, hydrochloric acid is molten
Liquid soaking time is 3-4h, which is to thoroughly remove and some present on Dispersed Cuticles can react not with hydrofluoric acid
The substance of molten object, the reaction time is too short to be will cause the carbonate mineral in step A-3 and reacts that form calcirm-fluoride attached with hydrofluoric acid
In cutin layer surface, cause the cuticula prepared of poor quality, be not used to subsequent observation analysis etc..
In addition, a large amount of silicate in Dispersed Cuticles uses since what is handled in the application is Dispersed Cuticles material
The hydrofluoric acid solution of 40%-45% is handled, and can thoroughly remove cuticula.
According to the preferred embodiment of the present invention, in stepb, cuticula oxidation treatment time is 3-12h.
According to the preferred embodiment of the present invention, in step C, the separation of the upper and lower epidermis of cuticula sample is in glass
On glass base station, has reeded glass plate centered on the glass base station.
According to the preferred embodiment of the present invention, the depth of the groove is 2-5mm, and the diameter of the groove is 1-2cm.
The present invention realizes phytolite dispersion angle in the preparation process of phytolite Dispersed Cuticles, using infusion method
The quick preparation of matter layer, after the oxidation processes of Shu Shi solution and potassium hydroxide treatment, mesophyll tissue carbonizes completely, because
And can easily separate cuticula, effectively shorten experimental period.And cuticula will not be destroyed in the treatment process,
The quality for improving the cuticula of preparation, the cutin layer surface free from admixture prepared, edge is not curled, is conducive to subsequent applications.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of inventive container.
Fig. 2 is the sectional view of inventive container.
Fig. 3 is the structural schematic diagram of inventive container lid.
Fig. 4 is the sectional view of inventive container.
Fig. 5 is the structural schematic diagram of glass base station of the present invention.
Fig. 6 is the sectional view of glass base station of the present invention.
Fig. 7 is the photo of 1 Dispersed Cuticles of embodiment under an optical microscope.
Fig. 8 is the photo of 1 Dispersed Cuticles of embodiment under a scanning electron microscope.
Fig. 9 is the photo of 1 Dispersed Cuticles of comparative example under an optical microscope.
Figure 10 is the photo of 1 Dispersed Cuticles of comparative example under a scanning electron microscope.
Specific embodiment
The present invention is described in detail with reference to embodiments, but the present invention is not limited by following embodiments.
1 Yanbian Area, Jilin Province Early Creataceous Epoch of embodiment long wealth group Dispersed Cuticles
The preparation method of phytolite Dispersed Cuticles, comprising:
Step A: Dispersed Cuticles sample pre-treatments:
(1) under the microscope using hand specimen, Dispersed Cuticles are chosen and save preferable carbonaceous mudstone sample, by cuticula and
Country rock is removed, and is put into the container sample slot of Fig. 1 in order, and is numbered in corresponding container cover；
(2) be separately added into distilled water to covering cuticula and country rock sample according to number, 40 DEG C heating water bath 45 minutes；
(3) distilled water and country rock clast in sample cell are removed, 35% hydrochloric acid solution is added, covered container lid impregnates sample
It product 3 hours, is then cleaned repeatedly with distilled water to neutrality；
(4) be added concentration be 40% hydrofluoric acid solution, covered container lid, 40 DEG C heating water bath 18 hours, until country rock is complete
Become cotton-shaped or powdered entirely, then cleaned up cotton-shaped or powdered country rock repeatedly with distilled water, until neutral；
Step B: Dispersed Cuticles oxidation processes:
(5) 1 part of saturation potassium chlorate solution is added in 2.5 parts of fuming nitric aicds, Shu Shi solution is made.It is cooling to Shu Shi solution
To room temperature, Shu Shi solution is added in sample and impregnates cuticula, covered container lid.Observation in real time, according to leaf film browning color half
Transparent or transparent sequence terminates reaction respectively, and is cleaned repeatedly with distilled water to neutrality, then uses distilled water immersion.
Step C: the upper and lower epidermis separation of Dispersed Cuticles:
(6) hydroxide of 1 drop (0.02mL) 5% was added with dropper with after distilled water immersion 1 hour every 2 minutes for sample
Potassium solution terminates reaction, is added until remaining mesophyll tissue and vascular bundle becomes to fall off after brown floccule or filiform to disperse
Distilled water is counter to be cleaned to neutrality；
(7) it moves to cuticula is careful on the groove of glass base station as shown in Figure 3, a small amount of distilled water is added, aobvious
The upper and lower epidermis of cuticula is separated with dissecting needle under micro mirror, and is removed the impurity on cuticula with hairbrush, according under number mirror
It takes pictures and differentiation and records upper lower epidermis and interior outside.Isolated cuticula cleans decontamination with distilled water repeatedly, again with distillation
Water, which impregnates, keeps cuticula wet, to observe to subsequent film-making.
Photo is as shown in FIG. 7 and 8 under photo and electron microscope under the optical microscopy of Dispersed Cuticles obtained.It can be seen that
Cuticula is complete, and surface can clearly obtain the features such as cuticula stomatal apparatus, cell, mastoid process without attachment.
1 Yanbian Area, Jilin Province Early Creataceous Epoch of comparative example long wealth group Dispersed Cuticles
Comparative example 1 is identical as the operating method of embodiment 1, and difference is only that in comparative example 1 distilled water immersion in step
With heating water bath unused during hydrofluoric acid dips, and then HCl treatment first is used with hydrofluoric acid treatment in comparative example 1.
Photo is as shown in Figures 9 and 10 under photo and electron microscope under the optical microscopy of Dispersed Cuticles obtained.It can
See that cuticula is broken, impurity is adhered on surface, influences the observing effects such as stomatal apparatus, cell.
Although hereinbefore having been made with reference to some embodiments, invention has been described, Ben Fafan is not being departed from
In the case where enclosing, various improvement can be carried out to it, exhaustive is not carried out to the case where these combinations in the present specification and is retouched
The considerations of stating merely for the sake of omitting length and economizing on resources.Therefore, the invention is not limited to particular implementations disclosed herein
Example, but fall into all technical solutions of the scope of the claims.
Claims (10)
1. a kind of preparation method of phytolite Dispersed Cuticles, comprising:
Step A: it chooses cuticula sample and removes the water-solubility impurity in sample, carbonate components and siliceous ingredient, before obtaining
Handle Dispersed Cuticles sample；
Step B: the pre-treatment Dispersed Cuticles sample that step A is obtained being placed in Shu Shi solution and is impregnated, until leaf film browning color half
Reaction is terminated when transparent or transparent, is washed to neutrality, is obtained the Dispersed Cuticles sample of oxidation processes；
Step C: after the Dispersed Cuticles samples with water by oxidation processes is soaked for a period of time, being added potassium hydroxide solution,
Reaction is terminated after becoming brown floccule or filiform to remaining mesophyll tissue and vascular bundle, is washed to neutrality, described in taking-up
Dispersed Cuticles sample, which is placed on glass base station, removes upper and lower epidermis, obtains cuticula；
Step D: removing the impurity on resulting cuticula sample and clean cuticula, is subsequently placed in water and impregnates to subsequent system
Piece observation.
2. according to claim 1-method, which is characterized in that the Shu Shi solution includes saturation potassium chlorate solution and Fa
Cigarette nitric acid, it is preferable that the volume ratio of the saturation potassium chlorate solution and fuming nitric aicd is 1:(2-3).
3. method according to claim 1 or 2, which is characterized in that in step A, remove water-solubility impurity with distilled water.
4. method according to any one of claim 1-3, which is characterized in that in step A, remove carbonate with acid solution
Ingredient, preferably hydrochloric acid solution.
5. method according to any of claims 1-4, which is characterized in that in step A, remove silicon with hydrofluoric acid solution
Matter ingredient.
6. method according to any one of claims 1-5, which is characterized in that in step A, before Dispersed Cuticles sample
Reason includes:
Step A-1, the cuticula sample of selection is placed in order in the container with multiple sample cells, adds distilled water immersion
Then the cuticula sample 40-60min removes the distilled water in sample cell；
Step A-2, hydrochloric acid solution is added and impregnates 3-4h, then hydrochloric acid is removed and washes the cuticula sample to neutrality；
Step A-3, hydrofluoric acid solution is added to impregnate 12-24 hours, until country rock becomes cotton-shaped or powdered completely, it will with distilled water
Cotton-shaped or powdered country rock is cleaned repeatedly to neutrality.
7. according to the method described in claim 6, it is characterized in that, in the step A-1 distilled water immersion at 40-60 DEG C into
In capable and/or step A-2 the volumetric concentration of hydrochloric acid be 35-40% and/or step A-3 described in the volumetric concentration of hydrofluoric acid be
40-45%, preferably hydrofluoric acid dips carry out at 40-60 DEG C in step A-3.
8. method according to any one of claims 1-7, which is characterized in that in stepb, when cuticula oxidation processes
Between be 3-12h.
9. method according to claim 1 to 8, which is characterized in that in step C, the cuticula sample
The separation of upper and lower epidermis carries out on base station, with the glass plate of groove centered on the base station is preferred.
10. according to the method described in claim 9, it is characterized in that, the depth of the groove be 2-5mm, the groove it is straight
Diameter is 1-2cm.
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| CN201710748834.4A CN109425521B (en) | 2017-08-28 | 2017-08-28 | Preparation method of fossil plant dispersed cuticle |
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| CN201710748834.4A CN109425521B (en) | 2017-08-28 | 2017-08-28 | Preparation method of fossil plant dispersed cuticle |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110686954A (en) * | 2019-11-12 | 2020-01-14 | 兰州大学 | Pretreatment method for extracting plant fossil cuticle |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100111855A1 (en) * | 2008-10-31 | 2010-05-06 | Searete Llc, A Limited Liability Corporation Of The States Of Delaware | Frozen compositions and methods for piercing a substrate |
| CN103073038A (en) * | 2013-01-23 | 2013-05-01 | 浙江大学 | Preparation method of acid-modified superfine shell powder and product |
| CN103329659A (en) * | 2013-06-06 | 2013-10-02 | 商丘师范学院 | Method for improving germination rate and survival rate of sophora alopecuroide seeds and sophora alopecuroide seeds |
| US20130296185A1 (en) * | 2004-03-31 | 2013-11-07 | Dermtech International | Tape stripping methods for analysis of skin disease and pathological skin state |
| CN104316526A (en) * | 2014-10-23 | 2015-01-28 | 南阳师范学院 | Method for observing morphological structure in mesophyll of fossil plant |
| CN106596605A (en) * | 2016-11-29 | 2017-04-26 | 中国石油天然气股份有限公司 | Method for preparing high purity kerogen |
-
2017
- 2017-08-28 CN CN201710748834.4A patent/CN109425521B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130296185A1 (en) * | 2004-03-31 | 2013-11-07 | Dermtech International | Tape stripping methods for analysis of skin disease and pathological skin state |
| US20100111855A1 (en) * | 2008-10-31 | 2010-05-06 | Searete Llc, A Limited Liability Corporation Of The States Of Delaware | Frozen compositions and methods for piercing a substrate |
| CN103073038A (en) * | 2013-01-23 | 2013-05-01 | 浙江大学 | Preparation method of acid-modified superfine shell powder and product |
| CN103329659A (en) * | 2013-06-06 | 2013-10-02 | 商丘师范学院 | Method for improving germination rate and survival rate of sophora alopecuroide seeds and sophora alopecuroide seeds |
| CN104316526A (en) * | 2014-10-23 | 2015-01-28 | 南阳师范学院 | Method for observing morphological structure in mesophyll of fossil plant |
| CN106596605A (en) * | 2016-11-29 | 2017-04-26 | 中国石油天然气股份有限公司 | Method for preparing high purity kerogen |
Non-Patent Citations (2)
| Title |
|---|
| 云飞: "云南腾冲上新世五种植物化石角质层研究", 《中国优秀硕士学位论文全文数据库 基础科学辑》 * |
| 马清温 等: "化石植物研究方法简介", 《生物学通报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110686954A (en) * | 2019-11-12 | 2020-01-14 | 兰州大学 | Pretreatment method for extracting plant fossil cuticle |
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