CN106802253A - The paraffin section method of meadowrueleaf corydalis root stem - Google Patents
The paraffin section method of meadowrueleaf corydalis root stem Download PDFInfo
- Publication number
- CN106802253A CN106802253A CN201611264733.1A CN201611264733A CN106802253A CN 106802253 A CN106802253 A CN 106802253A CN 201611264733 A CN201611264733 A CN 201611264733A CN 106802253 A CN106802253 A CN 106802253A
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- China
- Prior art keywords
- alcohol
- meadowrueleaf corydalis
- corydalis root
- volumetric concentration
- root stem
- Prior art date
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- 241000218176 Corydalis Species 0.000 title claims abstract description 83
- 239000012188 paraffin wax Substances 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 23
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 43
- RZSYLLSAWYUBPE-UHFFFAOYSA-L Fast green FCF Chemical compound [Na+].[Na+].C=1C=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C(=CC(O)=CC=2)S([O-])(=O)=O)C=CC=1N(CC)CC1=CC=CC(S([O-])(=O)=O)=C1 RZSYLLSAWYUBPE-UHFFFAOYSA-L 0.000 claims abstract description 22
- 238000000926 separation method Methods 0.000 claims abstract description 13
- 230000018044 dehydration Effects 0.000 claims abstract description 10
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 10
- 230000001476 alcoholic effect Effects 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 206
- 239000000243 solution Substances 0.000 claims description 64
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 36
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 36
- 238000004043 dyeing Methods 0.000 claims description 33
- 229960000583 acetic acid Drugs 0.000 claims description 15
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 12
- 235000006408 oxalic acid Nutrition 0.000 claims description 12
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 claims description 11
- 239000002904 solvent Substances 0.000 claims description 11
- 239000011259 mixed solution Substances 0.000 claims description 10
- 210000001367 artery Anatomy 0.000 claims description 5
- 239000000428 dust Substances 0.000 claims description 5
- 238000007667 floating Methods 0.000 claims description 5
- 239000012362 glacial acetic acid Substances 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 239000003086 colorant Substances 0.000 claims description 4
- 238000007654 immersion Methods 0.000 claims description 2
- 238000012545 processing Methods 0.000 claims description 2
- 238000002604 ultrasonography Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims 1
- 239000000463 material Substances 0.000 abstract description 27
- 239000000975 dye Substances 0.000 abstract description 8
- 239000001993 wax Substances 0.000 description 64
- 239000004575 stone Substances 0.000 description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 21
- 239000000123 paper Substances 0.000 description 9
- 235000002991 Coptis groenlandica Nutrition 0.000 description 6
- 244000247747 Coptis groenlandica Species 0.000 description 6
- 241001411320 Eriogonum inflatum Species 0.000 description 6
- 239000006059 cover glass Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000011521 glass Substances 0.000 description 6
- 238000007711 solidification Methods 0.000 description 6
- 230000008023 solidification Effects 0.000 description 6
- 239000011435 rock Substances 0.000 description 5
- 238000005086 pumping Methods 0.000 description 4
- 150000001298 alcohols Chemical class 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000001704 evaporation Methods 0.000 description 3
- 229920000159 gelatin Polymers 0.000 description 3
- 239000008273 gelatin Substances 0.000 description 3
- 235000019322 gelatine Nutrition 0.000 description 3
- 235000011852 gelatine desserts Nutrition 0.000 description 3
- 210000004932 little finger Anatomy 0.000 description 3
- 239000000155 melt Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 229920001206 natural gum Polymers 0.000 description 3
- 238000010422 painting Methods 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- -1 wherein Chemical compound 0.000 description 3
- 239000002253 acid Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 241000783421 Corydalis saxicola Species 0.000 description 1
- 241000876833 Emberizinae Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 206010017553 Furuncle Diseases 0.000 description 1
- 235000019738 Limestone Nutrition 0.000 description 1
- SJEYSFABYSGQBG-UHFFFAOYSA-M Patent blue Chemical compound [Na+].C1=CC(N(CC)CC)=CC=C1C(C=1C(=CC(=CC=1)S([O-])(=O)=O)S([O-])(=O)=O)=C1C=CC(=[N+](CC)CC)C=C1 SJEYSFABYSGQBG-UHFFFAOYSA-M 0.000 description 1
- 235000016551 Potentilla erecta Nutrition 0.000 description 1
- 240000000103 Potentilla erecta Species 0.000 description 1
- 229920002334 Spandex Polymers 0.000 description 1
- 239000000980 acid dye Substances 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 239000010977 jade Substances 0.000 description 1
- 239000006028 limestone Substances 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—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
-
- 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
Abstract
The invention discloses a kind of paraffin section method of meadowrueleaf corydalis root stem, fixed including sample, be dehydrated, transparent, waxdip, embedding, section with exhibition piece, dewaxing, dye and mounting, before transparent step, the meadowrueleaf corydalis root stem after dehydration is put in the sarranine solution that volumetric concentration is 0.8 1.2% and contaminates 8 12h；After dewaxing, the slide that meadowrueleaf corydalis root root will be loaded with carries out rehydration, being put in again after rehydration in alcoholic solution carries out color separation, it is simultaneously the acetic acid of 1/ (800 1000) to volume ratio is added in the alcoholic solution for color separation, is finally putting into the fast green solution that volumetric concentration is 0.8 1.2% and dyes 10 20 seconds.Meadowrueleaf corydalis root stem dicing method of the invention can cut out the meadowrueleaf corydalis root stem structure of clear in structure, and sample can for a long time be preserved and used, and be permanent microscope slide sample, meadowrueleaf corydalis root medicinal material microscopical characters application is available for, for standard market.
Description
Technical field
The present invention relates to histology field.It is more particularly related to a kind of paraffin section method of meadowrueleaf corydalis root stem.
Background technology
Meadowrueleaf corydalis root (Corydalis saxicola Bunting) is bloodroot, is herbaceos perennial, distribution
Saved in China Guangxi, Guangdong, Fujian, Hunan etc..All herbal medicine, herb contains deydrokaividing (Corydalis Saxicolae) isoreactivity composition；
With significant antibacterial, anti-inflammatory, analgesia and strong stable effect, and observe there is suppression tumour cell to act on；Cure mainly sore furuncle poison,
The diseases such as hepatitis, cirrhosis, liver cancer.It is famous strong precious jade medicine kind, meadowrueleaf corydalis root plant distributions are confined to Limestone Mountain Areas, belong to tor
Endemic species.Because habitat conditions is severe, its natural propagation rate is very low, and population development is difficult, and resource reserves are extremely limited, add
Its seed is tiny, breeds relatively difficult, more difficult in introducing and planting to survive, and people's largely excavation and purchase, result in open country over year
Supply falls short of demand in production-goods source, and wild meadowrueleaf corydalis root resource is on the verge of exhaustion.With the increasingly soaring market price, adulterant layer goes out not
Thoroughly, therefore it is badly in need of developing the section of meadowrueleaf corydalis root stem and is used to differentiate the true and false of medicinal material, for specification medicinal material market, the rock discerned the false from the genuine is yellow
Even section needs the effect for reaching for meadowrueleaf corydalis root stem piece cutting structure is clear, can clearly differentiate each institutional framework etc., and need mark
Originally can for a long time preserve and use, be permanent microscope slide sample.The numerous and diverse disunity of conventional section technical step, and prepare
Meadowrueleaf corydalis root stem section do not reach requirement cannot be applied to medicinal material discriminating.
The content of the invention
Meadowrueleaf corydalis root root section it is an object of the invention to solve prior art preparation not enough clearly cannot be used for medicine
The problem that the material true and false differentiates, and the advantage that at least will be described later is provided.
In order to realize these purposes of the invention and further advantage, there is provided a kind of paraffin section side of meadowrueleaf corydalis root stem
Method, including sample fix, be dehydrated, transparent, waxdip, embedding, section with exhibition piece, dewaxing, dye and mounting, including：Transparent
Before step, the meadowrueleaf corydalis root stem after dehydration is put in the sarranine solution that volumetric concentration is 0.8-1.2% and contaminates 8-12h；
After dewaxing, the slide that will be loaded with meadowrueleaf corydalis root root carries out rehydration, is put in again after rehydration in alcoholic solution and is divided
Color, while being the acetic acid of 1/ (800-1000) to volume ratio is added in the alcoholic solution for color separation, is finally putting into volumetric concentration
To be dyeed 10-20 seconds in the fast green solution of 0.8-1.2%.
Preferably, the volumetric concentration is that the sarranine solution of 0.8-1.2% is with the volume of 2/3 volume absolute alcohol+1/3
The mixed liquor of TO types biology clarifier is formulated for solvent, and the volumetric concentration is to use body for the fast green solution of 0.8-1.2%
Product concentration be 95% alcoholic solution for solvent is formulated.
Preferably, being loaded with the slide of meadowrueleaf corydalis root stem, to be sequentially placed into 1/2 volume type biology clarifier+1/2 volume anhydrous
Alcohol blend, absolute alcohol, absolute alcohol, volumetric concentration are 95% alcohol, volumetric concentration is 85% alcohol, volumetric concentration is
Rehydration in 70% alcohol, every grade 3-5 seconds；Place into color separation in 50% alcohol, 3-10 seconds；Being sequentially placed into volumetric concentration again afterwards is
70% alcohol, volumetric concentration be 85% alcohol in further process, every grade 3-10 seconds.
Preferably, the slide of meadowrueleaf corydalis root stem will be loaded with after the dyeing of fast green solution it be put in be put into absolute alcohol and washes away dyeing
Agent, this level is stopped 3-10 seconds；Then it is sequentially placed into absolute alcohol, the 1/2TO types biology absolute alcohol mixed liquor of clarifier+1/2,
Every grade stops 3-10 seconds；To finally be loaded with during the slide of meadowrueleaf corydalis root stem is put into TO types biology clarifier and stop 5-10 minutes, this step
Suddenly in triplicate.
Preferably, the meadowrueleaf corydalis root stem after fixation is sequentially passed through into volumetric concentration for 80% alcohol, volumetric concentration are 85%
Alcohol, volumetric concentration be 90% alcohol, volumetric concentration be 95% alcohol be dehydrated step by step, each concentration gradient dehydration 45-60 minutes,
It is put in again in 100% alcohol and is dehydrated twice, it is each 30-45 minutes.
Preferably, the alcoholic solution of each gradient concentration carries out vacuum suction simultaneously in meadowrueleaf corydalis root stem dehydration
10-15 minutes.
Preferably, the stem after being dyeed to sarranine solution is biological thoroughly with the volume TO types of 1/2 volume absolute alcohol+1/2 successively
Bright dose of mixed liquor and the volume TO types of 1/3 volume absolute alcohol+2/3 biology clarifier mixed liquor carry out transparent processing, and every grade thoroughly
The bright time is 60-120 minutes；It is put in again in pure TO types biology clarifier and soaks 2 times, every time immersion 60-120 minute.
Preferably, before fixed, removal meadowrueleaf corydalis root stem surface floating dust, it is 5-10mm long, 3- wide that vertical its middle arteries are crosscutting
The stem section of 5mm, then stem section is put into FAA fixers fixed, FAA fixers are 70% alcohol by volumetric concentration:Glacial acetic acid:First
Aldehyde volume ratio is 90:5:5 are formulated.
Preferably, in meadowrueleaf corydalis root stem sarranine dyeing course, to being placed with the 0.8- of meadowrueleaf corydalis root stem after dyeing the 1st hour
1.2% sarranine solution carried out with the ultrasonic wave that supersonic frequency is 32-35kHz it is ultrasonically treated 60-75 seconds, dyeing the 3rd hour after, it is right
The 0.8-1.2% sarranine solution for being placed with meadowrueleaf corydalis root stem carries out ultrasonically treated 45-60 with the ultrasonic wave that supersonic frequency is 28-31kHz
Second, after dyeing the 7th hour, the 0.8-1.2% sarranine solution supersonic frequencies to being placed with meadowrueleaf corydalis root stem are the ultrasound of 25-27kHz
Ripple carries out ultrasonically treated 30-50 seconds.
Preferably, for being separately added into volume ratio for 1/ (1000- in 70% alcohol and 85% alcohol of further treatment
1200) mixed solution of citric acid, picric acid and oxalic acid, wherein citric acid in mixed solution：Picric acid：The volume of oxalic acid
Than being 1:2:1.
The present invention at least includes following beneficial effect：
(1) meadowrueleaf corydalis root stem dicing method disclosed by the invention, meadowrueleaf corydalis root stem cross cut structure is clear, can be in 10 to 12 days
The meadowrueleaf corydalis root stem structure of clear in structure is cut out, sample can for a long time be preserved and used, and be permanent microscope slide sample, be available for rock yellow
Even medicinal material microscopical characters application, for standard market；
(2) meadowrueleaf corydalis root stem dehydration back root part is nearly transparent, and the present invention preceding is first carried out with sarranine solution transparent to sample
Pre-staining, so the position that embeds is more accurate when embedding, sarranine solution of the invention be with 2/3 volume absolute alcohol+
The mixed liquor of 1/3 volume TO types biology clarifier is formulated for solvent, so can both carry out it is transparent can also be dyeed,
It is time saving and energy saving；
(3) addition that the present invention in 50% alcohol adds acetic acid and carries out color separation acetic acid before fast green dyeing is conducive to point
Color so that the fast green dyeing different parts effect of sarranine becomes apparent from；
(4) after color separation with volumetric concentration be 70% alcohol, volumetric concentration be 85% alcohol to the further treatment of sample, can
More three-dimensional with each tissue in causing meadowrueleaf corydalis root root, effect is more prominent after dyeing, it is easier to observation contrast；
(5) after fast green dyeing, then by slice, thin piece be sequentially placed into absolute alcohol, 1/2TO types biology clarifier+1/2 it is anhydrous
In alcohol blend, every grade stops 3-10 seconds；To finally be loaded with during the slide of meadowrueleaf corydalis root blade is put into TO types biology clarifier and stop
Stay 5-10 minutes, this step is in triplicate, it is therefore an objective to remove remain in blade and slide waxing completely；
(6) when sarranine solution is contaminated meadowrueleaf corydalis root stem, ultrasonication is carried out to sarranine solution, can not only makes score
Son be decomposed it is smaller be easier to enter into meadowrueleaf corydalis root stem, due also to the effect of ultrasonic wave so that meadowrueleaf corydalis root stalk cell film is more
Easily resonance is consequently facilitating sarranine molecule is entered into cell membrane；
(7) before the dyeing of fast green solution, for being separately added into volume in 70% alcohol and 85% alcohol of further treatment
Than the mixed solution of citric acid, picric acid and oxalic acid for 1/ (1000-1200), not only cause that meadowrueleaf corydalis root stem structure is more vertical
Body, fast green dyestuff is acid dyes, and meadowrueleaf corydalis root stem is soaked with the acid of low concentration in advance, can cause that meadowrueleaf corydalis root stem is in
Acidity, more conducively fast green dyeing and color separation, citric acid and oxalic acid are the structure that natural acid will not destroy meadowrueleaf corydalis root stem, are added bitter
It is sour that further color and structure are fixed.
Further advantage of the invention, target and feature embody part by following explanation, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Brief description of the drawings
Fig. 1 is the paraffin section figure of control group of the invention；
Fig. 2 is the paraffin section figure of embodiments of the invention 1；
Fig. 3 is the paraffin section figure of embodiments of the invention 5.
Specific embodiment
The present invention is described in further detail below in conjunction with the accompanying drawings, to make those skilled in the art with reference to specification text
Word can be implemented according to this.
It should be noted that experimental technique described in following embodiments, unless otherwise specified, is conventional method, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
Embodiment 1：
(1) it is fixed：Fresh meadowrueleaf corydalis root stem adopt it is lower after wash down surface floating dust with running water, blot blade surface moisture.So
Vertical stem's middle arteries are crosscutting afterwards, and every is about 8mm, 4mm wide, are put into the FAA fixers that volumetric concentration is 70% alcohol
24 hours are fixed, FAA fixers are 70% alcohol by volumetric concentration:Glacial acetic acid:Formaldehyde volume ratio is 90:5:5 are formulated.
(2) it is dehydrated：Material is taken out from FAA fixers, filter paper blots unnecessary FAA fixers, is put into rubber stopper
In flat bottom glass pipe, volumetric concentration is added in 80% alcohol, volumetric concentration is that the addition of 80% alcohol is material volume
More than 20 times, vacuum suction 10 minutes is stopped 45 minutes；Volumetric concentration is poured out for 80% alcohol, it is 85% to add volumetric concentration
Alcohol, vacuum suction 10 minutes is placed 45 minutes；Volumetric concentration is poured out for 85% alcohol, addition volumetric concentration is 90% alcohol,
Vacuum suction 15 minutes, places 60 minutes；Volumetric concentration is poured out for 90% alcohol, addition volumetric concentration is 95% alcohol, vacuum
Pumping 15 minutes, places 60 minutes；Volumetric concentration is poured out for 95% alcohol, adds 100% alcohol, vacuum suction 15 minutes to put
Put 30 minutes；100% alcohol is changed, vacuum suction 15 minutes is placed 45 minutes.
(4) it is transparent：Absolute alcohol is poured out, before transparent, it is 1% that the meadowrueleaf corydalis root stem after dehydration is put in into volumetric concentration
Sarranine solution contaminates 10h, and wherein volumetric concentration is that 1% sarranine solution is given birth to the volume TO types of 2/3 volume absolute alcohol+1/3
The mixed liquor of thing clarifier is formulated for solvent；Sarranine solution is poured out, the life of the volume TO types of 1/2 volume absolute alcohol+1/2 is added
The mixed liquor of thing clarifier, vacuum suction 15 minutes is placed 60 minutes；Pour out the life of the volume TO types of 1/2 volume absolute alcohol+1/2
The mixed liquor of thing clarifier, adds the mixed liquor of the volume TO types of 1/3 volume absolute alcohol+2/3 biology clarifier, vacuum suction 15
Minute, place 90 minutes；The mixed liquor of the volume TO types of 1/3 volume absolute alcohol+2/3 biology clarifier is poured out, pure TO types are added
Biological clarifier, vacuum suction 15 minutes is placed 90 minutes；TO types biology clarifier is poured out, a TO types biology clarifier is changed,
Vacuum suction 15 minutes, places 120 minutes.
(5) waxdip：First time waxdip to the small wax stone of solid is added in the glass tube of muck coptis stem, for TO types give birth to by addition
The 1/3 of thing clarifier volume, is stoppered bottle stopper, is put into 45 DEG C of insulating boxs, after row adds same amount of wax stone again after wax thawing,
This step totally 5 times, untill wax no longer melts, last time plus wax open bottle stopper, allow TO clarifiers to volatilize as far as possible, this mistake
Journey needs 3 days.Second waxdip pours into crucible stem together with wax, then crucible moved into 60 DEG C of insulating boxs, solidification
Wax will be poured out after all dissolving together with stem, TO clarifiers, the paraffin for having melted that molten point is 56 DEG C be changed to, in 60 DEG C of insulating boxs
Middle placement 4h；Then the paraffin for having melted that molten point is 58 DEG C is changed to again, can embedding after placing 2h in 60 DEG C of insulating boxs.
(6) embed：The crucible for filling meadowrueleaf corydalis root stem is taken out from insulating box, Label clip is gone out to be placed on embedded box on one side, bottom
After wax slightly coagulates, dissecting needle is slightly warm on alcolhol burner to scratch the solidifying wax in carton surface, and the material in crucible is dialled in into embedded box immediately,
And put material well according to slice direction, stand cooling.In basin is put into after surface solidification, it is allowed to sink under water 30 minutes
Afterwards, embedded box is taken out, room temperature is dried, is in store for.
(7) section and exhibition piece：The set of wax stone and finishing：Wax stone is repaiied to suitably greatly by material size and cut direction
It is small.Sticky wax block：The small wax stone that will be fixed is adhered on wooden unit.Heated on alcolhol burner with scalpel, then contact wax stone bottom
Portion, softens bottom wax, is bonded on wooden unit rapidly.The position that wax stone is connected with wooden unit, should drip a little dewaxings, strengthen bonding
Fastness.The wooden unit of wax stone of adhering is fixed on slicer, required thickness (6-12 μm) is mixed up, is cut into slices, put in order
Put in ready big carton or on cardboard.With little finger painting very small amount gelatin bonding die agent on slide.Add on slide
Enough distilled water, slide is placed on 50 ± (1-2) DEG C of warm platform of exhibition.Wax band is divided into the small of suitable length by scalpel
Section, length dips in water less than the 1/5-2/5 of cover glass length with point of a knife, will stick up wax band, is placed on and is allowed on the water surface of slide
Rapid open and flat, adjustment wax band position is allowed to marshalling.Two pieces of paper on the warm platform other end pad of exhibition, after after the flattening of wax band, immediately will
Water on its slide is blotted only, then is put on warm platform；Water evaporating completely is treated, mark is engraved in section one end with masonry pen, be put into
Toasted in 45 DEG C of baking ovens and can just carry out next step after at least placing 1 day.
(8) dewax：The slice, thin piece that will be dyeed is sequentially placed into 3 bottles of TO types biology clarifiers, every grade 3 minutes, slough paraffin.
(9) dye：Slice, thin piece after dewaxing is sequentially placed into the 1/2 volume TO types biology volume absolute alcohol of clarifier+1/2
Mixed liquor, absolute alcohol, absolute alcohol, volumetric concentration are 95% alcohol, volumetric concentration is 85% alcohol, volumetric concentration is 70%
Rehydration in alcohol, every grade 3 seconds.It is color separation 10 seconds in 50% alcohol that slice, thin piece is put into volumetric concentration again, and volumetric concentration is 50% wine
Being added in smart solution has acetic acid, wherein, acetic acid：The volume ratio of 50% alcohol is 1:1000.Then slice, thin piece is sequentially placed into volume
Concentration be 70% alcohol, volumetric concentration be 85% alcohol in further process, every grade 7 seconds.Slice, thin piece is finally put into volumetric concentration
To be dyeed 18 seconds in 1% fast green solution, wherein volumetric concentration is that 1% fast green solution is with the alcohol that volumetric concentration is 95%
Solution is formulated for solvent.After the completion of dyeing, slice, thin piece is put into absolute alcohol, rinses the coloring agent on slice, thin piece, this level is stopped
Stay 10 seconds；Slice, thin piece is put into absolute alcohol again, is stopped 10 seconds；Slice, thin piece is put into 1/2 volume TO types biology clarifier+1/ afterwards
In the mixed liquor of 2 volume absolute alcohols, stop 3 seconds；Finally slice, thin piece is put into TO types biology clarifier, is stopped 5 minutes, this step
Suddenly in triplicate.
(10) mounting：Slice, thin piece is lain on paper, the TO types biology clarifier of bottom residual is wiped, 2-3 is dripped above and is added dropwise
Put on airs natural gum, cover glass dries on alcolhol burner, carefully covered down from side, it is to avoid bubble occurs, is put into 45 DEG C of baking ovens, dry
Can be taken off, post label, the information such as name title material, collecting location, time are noted on label.
Embodiment 2：
(1) it is fixed：Fresh meadowrueleaf corydalis root stem adopt it is lower after wash down surface floating dust with running water, blot blade surface moisture.So
Vertical root middle arteries are crosscutting afterwards, and every is about 5mm, 3mm wide, are put into the FAA fixers that volumetric concentration is 70% alcohol
24 hours are fixed, FAA fixers weight alcohol:Glacial acetic acid:Formaldehyde volume ratio is 90:5:5.
(2) it is dehydrated：Material is taken out from FAA fixers, filter paper blots unnecessary FAA fixers, is put into rubber stopper
In flat bottom glass pipe, volumetric concentration is added in 80% alcohol, volumetric concentration is that the addition of 80% alcohol is material volume
More than 20 times, vacuum suction 15 minutes is stopped 55 minutes；Volumetric concentration is poured out for 80% alcohol, it is 85% to add volumetric concentration
Alcohol, vacuum suction 10 minutes is placed 60 minutes；Volumetric concentration is poured out for 85% alcohol, addition volumetric concentration is 90% alcohol,
Vacuum suction 12 minutes, places 45 minutes；Volumetric concentration is poured out for 90% alcohol, addition volumetric concentration is 95% alcohol, vacuum
Pumping 15 minutes, places 45 minutes；Volumetric concentration is poured out for 95% alcohol, adds 100% alcohol, vacuum suction 10 minutes to put
Put 45 minutes；100% alcohol is changed, vacuum suction 15 minutes is placed 30 minutes.
(4) it is transparent：Absolute alcohol is poured out, before transparent, it is 0.8% that the meadowrueleaf corydalis root stem after dehydration is put in into volumetric concentration
Sarranine solution contaminate 12h, wherein volumetric concentration is that 0.8% sarranine solution is with the volume TO of 2/3 volume absolute alcohol+1/3
The mixed liquor of type biology clarifier is formulated for solvent；Sarranine solution is poured out, the volume TO of 1/2 volume absolute alcohol+1/2 is added
The mixed liquor of type biology clarifier, vacuum suction 10 minutes is placed 90 minutes；Pour out the volume TO of 1/2 volume absolute alcohol+1/2
The mixed liquor of type biology clarifier, adds the mixed liquor of the volume TO types of 1/3 volume absolute alcohol+2/3 biology clarifier, and vacuum is taken out
Gas 10 minutes, places 60 minutes；The mixed liquor of the volume TO types of 1/3 volume absolute alcohol+2/3 biology clarifier is poured out, is added pure
TO types biology clarifier, vacuum suction 10 minutes is placed 120 minutes；TO types biology clarifier is poured out, a TO type is changed biological thoroughly
Bright dose, vacuum suction 10 minutes is placed 60 minutes.
(5) waxdip：First time waxdip to the small wax stone of solid is added in the glass tube of muck coptis stem, for TO types give birth to by addition
The 1/3 of thing clarifier volume, is stoppered bottle stopper, is put into 45 DEG C of insulating boxs, after row adds same amount of wax stone again after wax thawing,
This step totally 3 times, untill wax no longer melts, last time plus wax open bottle stopper, allow TO clarifiers to volatilize as far as possible, this mistake
Journey needs 2 days.Second waxdip pours into crucible stem together with wax, then crucible moved into 60 DEG C of insulating boxs, solidification
Wax will be poured out after all dissolving together with stem, TO clarifiers, the paraffin for having melted that molten point is 56 DEG C be changed to, in 60 DEG C of insulating boxs
Middle placement 4h；Then the paraffin for having melted that molten point is 58 DEG C is changed to again, can embedding after placing 2h in 60 DEG C of insulating boxs.
(6) embed：The crucible for filling meadowrueleaf corydalis root stem is taken out from insulating box, Label clip is gone out to be placed on embedded box on one side, bottom
After wax slightly coagulates, dissecting needle is slightly warm on alcolhol burner to scratch the solidifying wax in carton surface, and the material in crucible is dialled in into embedded box immediately,
And put material well according to slice direction, stand cooling.In basin is put into after surface solidification, it is allowed to sink under water 30 minutes
Afterwards, embedded box is taken out, room temperature is dried, is in store for.
(7) section and exhibition piece：The set of wax stone and finishing：Wax stone is repaiied to suitably greatly by material size and cut direction
It is small.Sticky wax block：The small wax stone that will be fixed is adhered on wooden unit.Heated on alcolhol burner with scalpel, then contact wax stone bottom
Portion, softens bottom wax, is bonded on wooden unit rapidly.The position that wax stone is connected with wooden unit, should drip a little dewaxings, strengthen bonding
Fastness.The wooden unit of wax stone of adhering is fixed on slicer, required thickness (6-12 μm) is mixed up, is cut into slices, put in order
Put in ready big carton or on cardboard.With little finger painting very small amount gelatin bonding die agent on slide.Add on slide
Enough distilled water, slide is placed on 50 ± (1-2) DEG C of warm platform of exhibition.Wax band is divided into the small of suitable length by scalpel
Section, length dips in water less than the 1/5-2/5 of cover glass length with point of a knife, will stick up wax band, is placed on and is allowed on the water surface of slide
Rapid open and flat, adjustment wax band position is allowed to marshalling.Two pieces of paper on the warm platform other end pad of exhibition, after after the flattening of wax band, immediately will
Water on its slide is blotted only, then is put on warm platform；Water evaporating completely is treated, mark is engraved in section one end with masonry pen, be put into
Toasted in 45 DEG C of baking ovens and can just carry out next step after at least placing 2 days.
(8) dewax：The slice, thin piece that will be dyeed is sequentially placed into 3 bottles of TO types biology clarifiers, every grade 5 minutes, slough paraffin.
(9) dye：Slice, thin piece after dewaxing is sequentially placed into the 1/2 volume TO types biology volume absolute alcohol of clarifier+1/2
Mixed liquor, absolute alcohol, absolute alcohol, volumetric concentration are 95% alcohol, volumetric concentration is 85% alcohol, volumetric concentration is 70%
Rehydration in alcohol, every grade 5 seconds.It is color separation 3 seconds in 50% alcohol that slice, thin piece is put into volumetric concentration again, and volumetric concentration is 50% alcohol
Being added in solution has acetic acid, wherein, acetic acid：The volume ratio of 50% alcohol is 1:800.Then slice, thin piece is sequentially placed into volumetric concentration
Be 70% alcohol, volumetric concentration be 85% alcohol in further process, every grade 10 seconds.Slice, thin piece finally is put into volumetric concentration is
Dyeed 10 seconds in 1.2% fast green solution, wherein volumetric concentration is that 1.2% fast green solution is with the wine that volumetric concentration is 95%
Smart solution is formulated for solvent.After the completion of dyeing, slice, thin piece is put into absolute alcohol, rinses the coloring agent on slice, thin piece, this level
Stop 8 seconds；Slice, thin piece is put into absolute alcohol again, is stopped 3 seconds；Slice, thin piece is put into 1/2 volume TO types biology clarifier+1/ afterwards
In the mixed liquor of 2 volume absolute alcohols, stop 10 seconds；Finally slice, thin piece is put into TO types biology clarifier, is stopped 10 minutes, this
Step is in triplicate.
(10) mounting：Slice, thin piece is lain on paper, the TO types biology clarifier of bottom residual is wiped, 2-3 is dripped above and is added dropwise
Put on airs natural gum, cover glass dries on alcolhol burner, carefully covered down from side, it is to avoid bubble occurs, is put into 45 DEG C of baking ovens, dry
Can be taken off, post label, the information such as name title material, collecting location, time are noted on label.
Embodiment 3：
(1) it is fixed：Fresh meadowrueleaf corydalis root stem adopt it is lower after wash down surface floating dust with running water, blot blade surface moisture.Then
Vertical root middle arteries are crosscutting, and every is about 10mm, 5mm wide, are put into solid in the FAA fixers that volumetric concentration is 70% alcohol
It is fixed 24 hours, FAA fixers weight alcohol:Glacial acetic acid:Formaldehyde volume ratio is 90:5:5.
(2) it is dehydrated：Material is taken out from FAA fixers, filter paper blots unnecessary FAA fixers, is put into rubber stopper
In flat bottom glass pipe, volumetric concentration is added in 80% alcohol, volumetric concentration is that the addition of 80% alcohol is material volume
More than 20 times, vacuum suction 10 minutes is stopped 60 minutes；Volumetric concentration is poured out for 80% alcohol, it is 85% to add volumetric concentration
Alcohol, vacuum suction 10 minutes is placed 50 minutes；Volumetric concentration is poured out for 85% alcohol, addition volumetric concentration is 90% alcohol,
Vacuum suction 15 minutes, places 55 minutes；Volumetric concentration is poured out for 90% alcohol, addition volumetric concentration is 95% alcohol, vacuum
Pumping 15 minutes, places 55 minutes；Volumetric concentration is poured out for 95% alcohol, adds 100% alcohol, vacuum suction 15 minutes to put
Put 40 minutes；100% alcohol is changed, vacuum suction 10 minutes is placed 40 minutes.
(4) it is transparent：Absolute alcohol is poured out, before transparent, the meadowrueleaf corydalis root stem after dehydration is put in into volumetric concentration is
0.8% sarranine solution contaminates 12h, and wherein volumetric concentration is that 0.8% sarranine solution is with the body of 2/3 volume absolute alcohol+1/3
The mixed liquor of product TO type biology clarifiers is formulated for solvent；Sarranine solution is poured out, the body of 1/2 volume absolute alcohol+1/2 is added
The mixed liquor of product TO type biology clarifiers, vacuum suction 10 minutes is placed 120 minutes；Pour out the body of 1/2 volume absolute alcohol+1/2
The mixed liquor of product TO type biology clarifiers, adds the mixed liquor of the volume TO types of 1/3 volume absolute alcohol+2/3 biology clarifier, very
Sky pumping 15 minutes, places 120 clocks；The mixed liquor of the volume TO types of 1/3 volume absolute alcohol+2/3 biology clarifier is poured out, is added
Pure TO types biology clarifier, vacuum suction 15 minutes is placed 60 minutes；TO types biology clarifier is poured out, a TO type is changed biological
Clarifier, vacuum suction 15 minutes is placed 90 minutes.
(5) waxdip：First time waxdip to the small wax stone of solid is added in the glass tube of muck coptis stem, for TO types give birth to by addition
The 1/3 of thing clarifier volume, is stoppered bottle stopper, is put into 45 DEG C of insulating boxs, after row adds same amount of wax stone again after wax thawing,
This step totally 4 times, untill wax no longer melts, last time plus wax open bottle stopper, allow TO clarifiers to volatilize as far as possible, this mistake
Journey needs 2 days.Second waxdip pours into crucible stem together with wax, then crucible moved into 60 DEG C of insulating boxs, solidification
Wax will be poured out after all dissolving together with stem, TO clarifiers, the paraffin for having melted that molten point is 56 DEG C be changed to, in 60 DEG C of insulating boxs
Middle placement 4h；Then the paraffin for having melted that molten point is 58 DEG C is changed to again, can embedding after placing 2h in 60 DEG C of insulating boxs.
(6) embed：The crucible for filling meadowrueleaf corydalis root stem is taken out from insulating box, Label clip is gone out to be placed on embedded box on one side, bottom
After wax slightly coagulates, dissecting needle is slightly warm on alcolhol burner to scratch the solidifying wax in carton surface, and the material in crucible is dialled in into embedded box immediately,
And put material well according to slice direction, stand cooling.In basin is put into after surface solidification, it is allowed to sink under water 30 minutes
Afterwards, embedded box is taken out, room temperature is dried, is in store for.
(7) section and exhibition piece：The set of wax stone and finishing：Wax stone is repaiied to suitably greatly by material size and cut direction
It is small.Sticky wax block：The small wax stone that will be fixed is adhered on wooden unit.Heated on alcolhol burner with scalpel, then contact wax stone bottom
Portion, softens bottom wax, is bonded on wooden unit rapidly.The position that wax stone is connected with wooden unit, should drip a little dewaxings, strengthen bonding
Fastness.The wooden unit of wax stone of adhering is fixed on slicer, required thickness (6-12 μm) is mixed up, is cut into slices, put in order
Put in ready big carton or on cardboard.With little finger painting very small amount gelatin bonding die agent on slide.Add on slide
Enough distilled water, slide is placed on 50 ± (1-2) DEG C of warm platform of exhibition.Wax band is divided into the small of suitable length by scalpel
Section, length dips in water less than the 1/5-2/5 of cover glass length with point of a knife, will stick up wax band, is placed on and is allowed on the water surface of slide
Rapid open and flat, adjustment wax band position is allowed to marshalling.Two pieces of paper on the warm platform other end pad of exhibition, after after the flattening of wax band, immediately will
Water on its slide is blotted only, then is put on warm platform；Water evaporating completely is treated, mark is engraved in section one end with masonry pen, be put into
Toasted in 45 DEG C of baking ovens and can just carry out next step after at least placing 1 day.
(8) dewax：The slice, thin piece that will be dyeed is sequentially placed into 3 bottles of TO types biology clarifiers, every grade 2-5 minutes, slough stone
Wax.
(9) dye：Slice, thin piece after dewaxing is sequentially placed into the 1/2 volume TO types biology volume absolute alcohol of clarifier+1/2
Mixed liquor, absolute alcohol, absolute alcohol, volumetric concentration are 95% alcohol, volumetric concentration is 85% alcohol, volumetric concentration is 70%
Rehydration in alcohol, every grade 4 seconds.It is color separation 6 seconds in 50% alcohol that slice, thin piece is put into volumetric concentration again, and volumetric concentration is 50% alcohol
Being added in solution has acetic acid, wherein, acetic acid：The volume ratio of 50% alcohol is 1:900.Then slice, thin piece is sequentially placed into volumetric concentration
Be 70% alcohol, volumetric concentration be 85% alcohol in further process, every grade 3 seconds.Slice, thin piece finally is put into volumetric concentration is
Dyeed 20 seconds in 0.8% fast green solution, wherein volumetric concentration is that 0.8% fast green solution is with the wine that volumetric concentration is 95%
Smart solution is formulated for solvent.After the completion of dyeing, slice, thin piece is put into absolute alcohol, rinses the coloring agent on slice, thin piece, this level
Stop 3 seconds；Slice, thin piece is put into absolute alcohol again, is stopped 7 seconds；Slice, thin piece is put into 1/2 volume TO types biology clarifier+1/ afterwards
In the mixed liquor of 2 volume absolute alcohols, stop 7 seconds；Finally slice, thin piece is put into TO types biology clarifier, is stopped 6 minutes, this step
Suddenly in triplicate.
(10) mounting：Slice, thin piece is lain on paper, the TO types biology clarifier of bottom residual is wiped, 2-3 is dripped above and is added dropwise
Put on airs natural gum, cover glass dries on alcolhol burner, carefully covered down from side, it is to avoid bubble occurs, is put into 45 DEG C of baking ovens, dry
Can be taken off, post label, the information such as name title material, collecting location, time are noted on label.
Embodiment 4：
On the basis of embodiment 1, in meadowrueleaf corydalis root stem sarranine dyeing course, to being placed with meadowrueleaf corydalis root after dyeing the 1st hour
The sarranine solution of stem is carried out ultrasonically treated 75 seconds with the ultrasonic wave that supersonic frequency is 32kHz, after dyeing the 3rd hour, to being placed with rock
The sarranine solution of coptis stem is carried out ultrasonically treated 45 seconds with the ultrasonic wave that supersonic frequency is 31kHz, after dyeing the 7th hour, to putting
The sarranine solution for having meadowrueleaf corydalis root stem is carried out ultrasonically treated 50 seconds with the ultrasonic wave that supersonic frequency is 25kHz, continues to contaminate to terminating.
Before the dyeing of fast green solution, it is for being separately added into volume ratio in 70% alcohol and 85% alcohol of further treatment
The mixed solution of 1/1000 citric acid, picric acid and oxalic acid, wherein citric acid in mixed solution：Picric acid：The body of oxalic acid
Product is than being 1:2:1.
Embodiment 5：
On the basis of embodiment 1, in meadowrueleaf corydalis root stem sarranine dyeing course, to being placed with meadowrueleaf corydalis root after dyeing the 1st hour
The sarranine solution of stem is carried out ultrasonically treated 60 seconds with the ultrasonic wave that supersonic frequency is 35kHz, after dyeing the 3rd hour, to being placed with rock
The sarranine solution of coptis stem is carried out ultrasonically treated 50 seconds with the ultrasonic wave that supersonic frequency is 30kHz, after dyeing the 7th hour, to putting
The sarranine solution for having meadowrueleaf corydalis root stem is carried out ultrasonically treated 30 seconds with the ultrasonic wave that supersonic frequency is 27kHz.
Before the dyeing of fast green solution, it is for being separately added into volume ratio in 70% alcohol and 85% alcohol of further treatment
The mixed solution of 1/1200 citric acid, picric acid and oxalic acid, wherein citric acid in mixed solution：Picric acid：The body of oxalic acid
Product is than being 1:2:1.
Embodiment 6：
On the basis of embodiment 1, in meadowrueleaf corydalis root stem sarranine dyeing course, to being placed with meadowrueleaf corydalis root after dyeing the 1st hour
The sarranine solution of stem is carried out ultrasonically treated 65 seconds with the ultrasonic wave that supersonic frequency is 33kHz, after dyeing the 3rd hour, to being placed with rock
The sarranine solution of coptis stem is carried out ultrasonically treated 60 seconds with the ultrasonic wave that supersonic frequency is 28kHz, after dyeing the 7th hour, to putting
The sarranine solution for having meadowrueleaf corydalis root stem is carried out ultrasonically treated 45 seconds with the ultrasonic wave that supersonic frequency is 26kHz.
Before the dyeing of fast green solution, it is for being separately added into volume ratio in 70% alcohol and 85% alcohol of further treatment
The mixed solution of 1/1100 citric acid, picric acid and oxalic acid, wherein citric acid in mixed solution：Picric acid：The body of oxalic acid
Product is than being 1:2:1.
In order to illustrate effect of the invention, the conventional paraffin section method of applicant makes the paraffin section conduct of stem
Control group, then Example 1 and stem's paraffin section obtained in embodiment 5, three is put in Lycra biomicroscope respectively
DM2000 observations are taken pictures, and respectively obtain Fig. 1, Fig. 2 and Fig. 3, and as seen from the figure, the paraffin section institutional framework of Fig. 3 is clear
Clearly, can be used for differentiating the true and false of medicinal material；Fig. 2 takes second place；The root tissue of Fig. 1 is less clear, and Fig. 1 cannot be used for differentiating medicinal material
The true and false.
Although embodiment of the present invention is disclosed as above, it is not restricted to listed in specification and implementation method
With, it can be applied to various suitable the field of the invention completely, for those skilled in the art, can be easily
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, the present invention is not limited
In specific details and shown here as the legend with description.
Claims (10)
1. a kind of paraffin section method of meadowrueleaf corydalis root stem, including sample is fixed, is dehydrated, transparent, waxdip, embedding, section with exhibition piece,
Dewaxing, dyeing and mounting, it is characterised in that
Before transparent step, the meadowrueleaf corydalis root stem after dehydration is put in the sarranine solution that volumetric concentration is 0.8-1.2% and contaminates 8-
12h；
After dewaxing, the slide that will be loaded with meadowrueleaf corydalis root root carries out rehydration, and being put in again after rehydration in alcoholic solution carries out color separation, together
When to volume ratio is added in the alcoholic solution for color separation be the acetic acid of 1/ (800-1000), be finally putting into volumetric concentration for 0.8-
Dyeed 10-20 seconds in 1.2% fast green solution.
2. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 1, it is characterised in that the volumetric concentration is 0.8-
1.2% sarranine solution be with the volume TO types of 2/3 volume absolute alcohol+1/3 biology clarifier mixed liquor be solvent prepare and
Into, the volumetric concentration for the fast green solution of 0.8-1.2% be with the alcoholic solution that volumetric concentration is 95% be solvent prepare and
Into.
3. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 1, it is characterised in that the slide of meadowrueleaf corydalis root stem will be loaded with
It is sequentially placed into the 1/2 volume type biology volume absolute alcohol of clarifier+1/2 mixed liquor, absolute alcohol, absolute alcohol, volumetric concentration
For 95% alcohol, volumetric concentration be 85% alcohol, volumetric concentration be 70% alcohol in rehydration, every grade 3-5 seconds；Place into 50% wine
Color separation in essence, 3-10 seconds；Volumetric concentration is sequentially placed into again afterwards for 70% alcohol, volumetric concentration further to locate in 85% alcohol
Reason, every grade 3-10 seconds.
4. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 1, it is characterised in that will be loaded with after fast green solution dyeing
The slide of meadowrueleaf corydalis root stem is put in be put into absolute alcohol and washes away coloring agent, and this level is stopped 3-10 seconds；Then it is sequentially placed into without watery wine
In essence, the 1/2TO types biology absolute alcohol mixed liquor of clarifier+1/2, every grade stops 3-10 seconds；To finally meadowrueleaf corydalis root stem be loaded with
Slide is stopped 5-10 minutes in being put into TO types biology clarifier, and this step is in triplicate.
5. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 1, it is characterised in that by the meadowrueleaf corydalis root stem after fixation according to
It is secondary by volumetric concentration be 80% alcohol, volumetric concentration be 85% alcohol, volumetric concentration be 90% alcohol, volumetric concentration be 95%
Alcohol is dehydrated step by step, and each concentration gradient is dehydrated 45-60 minute, then is put in 100% alcohol and is dehydrated twice, and each 30-45 divides
Clock.
6. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 5, it is characterised in that the alcohol of each gradient concentration is molten
Liquid is carried out vacuum suction 10-15 minutes simultaneously in meadowrueleaf corydalis root stem dehydration.
7. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 1, it is characterised in that the stem after being dyeed to sarranine solution
Successively with the volume TO types of 1/2 volume absolute alcohol+1/2 biology clarifier mixed liquor and the volume of 1/3 volume absolute alcohol+2/3
TO types biology clarifier mixed liquor carries out transparent processing, and every grade of clearing time is 60-120 minutes；It is biological that pure TO types are put in again
Soaked 2 times in clarifier, every time immersion 60-120 minutes.
8. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 1, it is characterised in that before fixed, removes meadowrueleaf corydalis root
Stem surface floating dust, it is 5-10mm long that vertical its middle arteries are crosscutting, the stem section of 3-5mm wide, then stem section is put into solid in FAA fixers
Fixed, FAA fixers are 70% alcohol by volumetric concentration:Glacial acetic acid:Formaldehyde volume ratio is 90:5:5 are formulated.
9. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 8, it is characterised in that dyeed in meadowrueleaf corydalis root stem sarranine
It is the ultrasound of 32-35kHz to the 0.8-1.2% sarranine solution supersonic frequencies that are placed with meadowrueleaf corydalis root stem after Cheng Zhong, dyeing the 1st hour
Ripple carry out it is ultrasonically treated 60-75 seconds, dyeing the 3rd hour after, the 0.8-1.2% sarranine solution supersonic frequencies to being placed with meadowrueleaf corydalis root stem
Rate for 28-31kHz ultrasonic wave carry out it is ultrasonically treated 45-60 second, dye the 7th hour after, the 0.8- to being placed with meadowrueleaf corydalis root stem
1.2% sarranine solution is carried out ultrasonically treated 30-50 seconds with the ultrasonic wave that supersonic frequency is 25-27kHz.
10. the paraffin section method of meadowrueleaf corydalis root stem as claimed in claim 3, it is characterised in that for further treatment
It is the mixed of the citric acid of 1/ (1000-1200), picric acid and oxalic acid that volume ratio is separately added into 70% alcohol and 85% alcohol
Close solution, wherein citric acid in mixed solution：Picric acid：The volume ratio of oxalic acid is 1:2:1.
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| CN109374376A (en) * | 2018-11-09 | 2019-02-22 | 上海市农业科学院 | A kind of slice preparation method suitable for mushroom lamella Basidium morphologic observation |
| CN110553890A (en) * | 2019-10-16 | 2019-12-10 | 甘肃农业大学 | safe and efficient paraffin slicing method for potato roots and leaves |
| CN113933136A (en) * | 2021-10-15 | 2022-01-14 | 华北理工大学 | Dendritic crystal corrosion reagent of medical Al-free zinc-based alloy, preparation method and use method |
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| CN106802253B (en) | 2019-07-30 |
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