CN106802253B - The paraffin section method of meadowrueleaf corydalis root stem - Google Patents
The paraffin section method of meadowrueleaf corydalis root stem Download PDFInfo
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- CN106802253B CN106802253B CN201611264733.1A CN201611264733A CN106802253B CN 106802253 B CN106802253 B CN 106802253B CN 201611264733 A CN201611264733 A CN 201611264733A CN 106802253 B CN106802253 B CN 106802253B
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- alcohol
- volumetric concentration
- meadowrueleaf corydalis
- corydalis root
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- 241000218176 Corydalis Species 0.000 title claims abstract description 76
- 239000012188 paraffin wax Substances 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 12
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 43
- 238000004043 dyeing Methods 0.000 claims abstract description 39
- 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 12
- 230000001476 alcoholic effect Effects 0.000 claims abstract description 9
- 239000000975 dye Substances 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 205
- 239000000243 solution Substances 0.000 claims description 63
- 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 33
- 229960000583 acetic acid Drugs 0.000 claims description 15
- 238000009210 therapy by ultrasound Methods 0.000 claims description 14
- 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
- 239000011259 mixed solution Substances 0.000 claims description 11
- 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
- 238000012545 processing Methods 0.000 claims description 9
- 210000001367 artery Anatomy 0.000 claims description 5
- 239000000428 dust Substances 0.000 claims description 5
- 239000012362 glacial acetic acid Substances 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 239000003086 colorant Substances 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 3
- 230000018044 dehydration Effects 0.000 claims description 3
- 238000006297 dehydration reaction Methods 0.000 claims description 3
- 244000248349 Citrus limon Species 0.000 claims 1
- 235000005979 Citrus limon Nutrition 0.000 claims 1
- 239000000463 material Substances 0.000 abstract description 27
- 230000007774 longterm Effects 0.000 abstract description 3
- 238000004321 preservation Methods 0.000 abstract description 3
- 239000001993 wax Substances 0.000 description 64
- 239000004575 stone Substances 0.000 description 25
- 239000011521 glass Substances 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 239000000123 paper Substances 0.000 description 9
- 230000000694 effects Effects 0.000 description 7
- 241001411320 Eriogonum inflatum Species 0.000 description 6
- 241000207929 Scutellaria Species 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
- 239000007787 solid Substances 0.000 description 4
- 108010010803 Gelatin Proteins 0.000 description 3
- 150000001298 alcohols Chemical class 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
- 229920001206 natural gum Polymers 0.000 description 3
- 238000005086 pumping Methods 0.000 description 3
- 238000006748 scratching Methods 0.000 description 3
- 230000002393 scratching effect Effects 0.000 description 3
- 239000008399 tap water Substances 0.000 description 3
- 235000020679 tap water Nutrition 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- 238000009826 distribution Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000000203 mixture 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
- 229920002334 Spandex Polymers 0.000 description 1
- 241000372442 Wachendorfia thyrsiflora Species 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
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding 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
- 229940079593 drug Drugs 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
- 239000004615 ingredient Substances 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
- 238000012986 modification Methods 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000004759 spandex Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 210000004158 stalk cell Anatomy 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000004018 waxing Methods 0.000 description 1
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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention discloses a kind of paraffin section methods of meadowrueleaf corydalis root stem, it fixes, be dehydrated including sample, is transparent, waxdip, embedding, slice and exhibition piece, dewaxing, dyeing and mounting, before transparent step, dewatered meadowrueleaf corydalis root stem is put in the sarranine solution that volumetric concentration is 0.8-1.2% and disseminates 8-12h；After dewaxing, the slide for being loaded with meadowrueleaf corydalis root root is subjected to rehydration, it is put in alcoholic solution again after rehydration and carries out color separation, the acetic acid that volume ratio is 1/ (800-1000) is added into the alcoholic solution for color separation simultaneously, 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 meadowrueleaf corydalis root stem structure clear in structure, and sample can be used with long-term preservation, be permanent microscope slide sample, for meadowrueleaf corydalis root medicinal material microscopical characters application, for standard market.
Description
Technical field
The present invention relates to histology fields.It is more particularly related to a kind of paraffin section method of meadowrueleaf corydalis root stem.
Background technique
Meadowrueleaf corydalis root (Corydalis saxicola Bunting) is bloodroot, is herbaceos perennial, distribution
It is saved in China Guangxi, Guangdong, Fujian, Hunan etc..All herbal medicine, herb contain deydrokaividing (Corydalis Saxicolae) isoreactivity ingredient；
With significant antibacterial, anti-inflammatory, analgesia and stable effect by force, and observes and inhibited tumour cell effect；Cure mainly sore furuncle poison,
The diseases such as hepatitis, cirrhosis, liver cancer.It is famous strong precious jade drug kind, meadowrueleaf corydalis root plant distributions are confined to Limestone Mountain Areas, belong to tor
Endemic species.Since habitat conditions is severe, natural propagation rate is very low, and population development is difficult, and resource reserves are extremely limited, in addition
Its seed is tiny, and breeding is relatively difficult, and more difficult in introducing and planting to survive, people largely excavate and purchase over year, result in open country
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 meadowrueleaf corydalis root stem and be sliced the true and false to identify medicinal material, for specification medicinal material market, the rock Huang discerned the false from the genuine
It is that meadowrueleaf corydalis root stem piece cutting structure is clear that even slice, which needs effect to be achieved, can clearly differentiate each institutional framework etc., and need to mark
Originally it can be used with long-term preservation, be permanent microscope slide sample.The many and diverse disunity of conventional section technical step, and prepare
Meadowrueleaf corydalis root stem slice do not reach requirement can not be suitable for medicinal material identification.
Summary of the invention
It is an object of the invention to solve the meadowrueleaf corydalis root root of prior art preparation slice not enough clearly to cannot be used for medicine
The problem of material true and false identifies, and the advantages of at least will be described later, is provided.
In order to realize these purposes and other advantages according to the present invention, a kind of paraffin section side of meadowrueleaf corydalis root stem is provided
Method, including sample is fixed, is dehydrated, is transparent, waxdip, embedding, slice and exhibition piece, dewaxing, dyeing and mounting, comprising: transparent
Before step, dewatered meadowrueleaf corydalis root stem is put in the sarranine solution that volumetric concentration is 0.8-1.2% and disseminates 8-12h；
After dewaxing, the slide for being loaded with meadowrueleaf corydalis root root is subjected to rehydration, is put in alcoholic solution and is divided again after rehydration
Color, while the acetic acid that volume ratio is 1/ (800-1000) being added into the alcoholic solution for color separation, it is finally putting into volumetric concentration
To be dyed 10-20 seconds in the fast green solution of 0.8-1.2%.
Preferably, it is with+1/3 volume of 2/3 volume absolute alcohol that the volumetric concentration, which is the sarranine solution of 0.8-1.2%,
The mixed liquor of TO type biology clarifier is formulated for solvent, and the volumetric concentration is that the fast green solution of 0.8-1.2% is to use body
The alcoholic solution that product concentration is 95% is that solvent is formulated.
Preferably, that the slide for being loaded with meadowrueleaf corydalis root stem is sequentially placed into 1/2 volume type biology+1/2 volume of clarifier is 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 later is
70% alcohol, volumetric concentration be 85% alcohol in be further processed, every grade 3-10 seconds.
Preferably, the slide for being loaded with meadowrueleaf corydalis root stem is put in be put into absolute alcohol after fast green solution dyeing and washes away dyeing
Agent, this grade stop 3-10 seconds；Then it is sequentially placed into absolute alcohol, in+1/2 absolute alcohol mixed liquor of 1/2TO type biology clarifier,
Every grade stop 3-10 seconds；Finally the slide for being loaded with meadowrueleaf corydalis root stem is put into TO type biology clarifier and is stopped 5-10 minutes, this step
Suddenly in triplicate.
Preferably, the meadowrueleaf corydalis root stem after fixation is successively passed through volumetric concentration is 80% alcohol, volumetric concentration 85%
Alcohol, volumetric concentration are 90% alcohol, volumetric concentration is that 95% alcohol is dehydrated step by step, and each concentration gradient is dehydrated 45-60 minutes,
It is put in 100% alcohol and is dehydrated twice again, it is 30-45 minutes each.
Preferably, the alcoholic solution of each gradient concentration is evacuated simultaneously in meadowrueleaf corydalis root stem dehydration
10-15 minutes.
Preferably ,+1/2 volume TO type biology of 1/2 volume absolute alcohol is successively used thoroughly to the stem after the dyeing of sarranine solution
Bright dose of mixed liquor and+2/3 volume TO type biology clarifier mixed liquor of 1/3 volume absolute alcohol carry out transparent processing, and every grade thoroughly
The bright time is 60-120 minutes；It is put in pure TO type biology clarifier and impregnates 2 times again, impregnate 60-120 minutes every time.
Preferably, before fixed, meadowrueleaf corydalis root stem surface floating dust is removed, its crosscutting vertical middle arteries are long 5-10mm, wide 3-
The stem section of 5mm, then stem section is put into FAA fixer and is fixed, FAA fixer is 70% alcohol: glacial acetic acid: first by volumetric concentration
Aldehyde volume ratio is formulated for 90:5:5.
Preferably, in meadowrueleaf corydalis root stem sarranine dyeing course, to the 0.8- for being placed with meadowrueleaf corydalis root stem after dyeing the 1st hour
1.2% sarranine solution carries out ultrasonic treatment 60-75 seconds with the ultrasonic wave that supersonic frequency is 32-35kHz, right after dyeing the 3rd hour
The 0.8-1.2% sarranine solution for being placed with meadowrueleaf corydalis root stem carries out ultrasonic treatment 45-60 with the ultrasonic wave that supersonic frequency is 28-31kHz
Second, after dyeing the 7th hour, the ultrasound for being 25-27kHz with supersonic frequency to the 0.8-1.2% sarranine solution for being placed with meadowrueleaf corydalis root stem
Wave carries out ultrasonic treatment 30-50 seconds.
Preferably, it is 1/ (1000- that volume ratio is separately added into 70% alcohol and 85% alcohol for further processing
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 for 1:2:1.
The present invention is include at least the following beneficial effects:
(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
Meadowrueleaf corydalis root stem structure clear in structure is cut out, it is permanent microscope slide sample that sample can be used with long-term preservation, for rock Huang
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 first carries out sample with sarranine solution transparent
Pre-staining, the position that embeds is more acurrate when embedding in this way, sarranine solution of the invention be with 2/3 volume absolute alcohol+
The mixed liquor of 1/3 volume TO type biology clarifier is formulated for solvent, can both carry out transparent or dyed in this way,
It is time saving and energy saving；
(3) present invention, which is added acetic acid in 50% alcohol before fast green dyeing and carries out the addition of color separation acetic acid, is conducive to point
Color, so that the fast green dyeing different parts effect of sarranine becomes apparent from；
(4) be 70% alcohol with volumetric concentration after color separation, volumetric concentration is that 85% alcohol is further processed sample, can
So that respectively organizing more three-dimensional in meadowrueleaf corydalis root root, effect is more prominent after dyeing, it is easier to observation comparison；
(5) in fast green dyeing and then that piece is sequentially placed into absolute alcohol, 1/2TO type biology clarifier+1/2 is anhydrous
In alcohol blend, every grade stop 3-10 seconds；Finally the slide for being loaded with meadowrueleaf corydalis root blade is put into TO type biology clarifier and is stopped
It stays 5-10 minutes, this step is in triplicate, it is therefore an objective to completely remove and remain in blade and slide waxing；
(6) when sarranine solution disseminates meadowrueleaf corydalis root stem, ultrasonication is carried out to sarranine solution, can not only be made point
Son be decomposed it is smaller be easier to enter in meadowrueleaf corydalis root stem, due also to the effect of ultrasonic wave, so that meadowrueleaf corydalis root stalk cell film is more
Resonance is easy consequently facilitating sarranine molecule enters in cell membrane；
(7) before the dyeing of fast green solution, volume is separately added into 70% alcohol and 85% alcohol for further processing
Than the mixed solution for the citric acid of 1/ (1000-1200), picric acid and oxalic acid, not only make meadowrueleaf corydalis root stem structure more vertical
Body, fast green dyestuff are acid dyes, impregnate in advance to the acid of meadowrueleaf corydalis root stem low concentration, meadowrueleaf corydalis root stem can be made to be 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, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Detailed description of the invention
Fig. 1 is the paraffin section figure of control group of the invention；
Fig. 2 is the paraffin section figure of the embodiment of the present invention 1；
Fig. 3 is the paraffin section figure of the embodiment of the present invention 5.
Specific embodiment
Present invention will be described in further detail below with reference to the accompanying drawings, to enable those skilled in the art referring to specification text
Word can be implemented accordingly.
It should be noted that experimental method described in following embodiments is unless otherwise specified conventional method, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
Embodiment 1:
(1) fixed: fresh meadowrueleaf corydalis root stem adopt it is lower after with tap water wash down surface floating dust, blot blade surface moisture.So
Vertical stem's middle arteries are crosscutting afterwards, and every is about 8mm, wide 4mm, are put into the FAA fixer that volumetric concentration is 70% alcohol
24 hours are fixed, FAA fixer is 70% alcohol by volumetric concentration: glacial acetic acid: formaldehyde volume ratio is formulated for 90:5:5.
(2) it is dehydrated: material being taken out from FAA fixer, filter paper blots extra FAA fixer, is put into rubber stopper
In flat bottom glass pipe, it is in 80% alcohol that volumetric concentration, which is added, and volumetric concentration is that the additional amount of 80% alcohol is material volume
It 20 times or more, vacuum suction 10 minutes, stops 45 minutes；Pouring out volumetric concentration is 80% alcohol, and it is 85% that volumetric concentration, which is added,
Alcohol vacuum suction 10 minutes, is placed 45 minutes；Pouring out volumetric concentration is 85% alcohol, and addition volumetric concentration is 90% alcohol,
It vacuum suction 15 minutes, places 60 minutes；Pouring out volumetric concentration is 90% alcohol, and addition volumetric concentration is 95% alcohol, vacuum
Pumping 15 minutes is placed 60 minutes；Pouring out volumetric concentration is 95% alcohol, and 100% alcohol is added, vacuum suction 15 minutes, puts
It sets 30 minutes；100% alcohol is replaced, vacuum suction 15 minutes, is placed 45 minutes.
(4) transparent: to pour out absolute alcohol, before transparent, it is 1% that dewatered meadowrueleaf corydalis root stem, which is put in volumetric concentration,
Sarranine solution disseminates 10h, and the sarranine solution that wherein volumetric concentration is 1% is raw with+1/3 volume TO type of 2/3 volume absolute alcohol
The mixed liquor of object clarifier is formulated for solvent；Sarranine solution is poured out, it is raw that+1/2 volume TO type of 1/2 volume absolute alcohol is added
The mixed liquor of object clarifier vacuum suction 15 minutes, is placed 60 minutes；It is raw to pour out+1/2 volume TO type of 1/2 volume absolute alcohol
The mixed liquor of+2/3 volume TO type biology clarifier of 1/3 volume absolute alcohol, vacuum suction 15 is added in the mixed liquor of object clarifier
Minute, it places 90 minutes；The mixed liquor for pouring out+2/3 volume TO type biology clarifier of 1/3 volume absolute alcohol, is added pure TO type
Biological clarifier vacuum suction 15 minutes, is placed 90 minutes；TO type biology clarifier is poured out, a TO type biology clarifier is changed,
It vacuum suction 15 minutes, places 120 minutes.
(5) waxdip: the small wax stone of solid is added into the glass tube of muck scutellaria for first time waxdip, and additional amount is raw for TO type
The 1/3 of object clarifier volume, is stoppered bottle stopper, is put into 45 DEG C of insulating boxs, and same amount of wax stone is added in row again after wax thawing,
This step totally 5 times, until wax no longer melts, last time plus wax open bottle stopper, allow TO clarifier to volatilize as far as possible, this mistake
Journey needs 3 days.Second of waxdip pours into stem in crucible together with wax, then crucible is moved in 60 DEG C of insulating boxs, solidification
Wax will be poured out after all dissolving together with stem, TO clarifier, and the paraffin that molten point is 56 DEG C has been melted in change, in 60 DEG C of insulating boxs
Middle placement 4h；Then it changes to again and has melted the paraffin that molten point is 58 DEG C, can be embedded after placing 2h in 60 DEG C of insulating boxs.
(6) it embeds: taking out the crucible for filling meadowrueleaf corydalis root stem from insulating box, Label clip is gone out to be placed on embedded box on one side, bottom
After wax slightly coagulates, wax is coagulated on dissecting needle slightly hot carton surface of scratching on alcolhol burner, and the material in crucible is dialled in embedded box immediately,
And put material according to slice direction well, stand cooling.It is put into basin after surface solidification, is allowed to sink under water 30 minutes
Afterwards, embedded box is taken out, room temperature is dried, is in store for.
(7) it is sliced and exhibition piece: the fixation of wax stone and finishing: wax stone being repaired by material size and cut direction to appropriate big
It is small.Sticky wax block: the small wax stone fixed is adhered on wooden unit.It is heated on alcolhol burner with scalpel, then contacts wax stone bottom
Portion softens bottom wax, is sticked on wooden unit rapidly.The position that wax stone is connect with wooden unit should drip a little dewaxings, reinforce bonding
Fastness.The wooden unit for wax stone of adhering is fixed on slicer, required thickness (6-12 μm) is mixed up, is sliced, is put in order
It sets in ready big carton or on cardboard.The bonding die agent of minute quantity gelatin is applied on glass slide with little finger.Add on slide
Glass slide is placed on the warm platform of 50 ± (1-2) DEG C of exhibition by enough distilled water.Wax band is divided into the small of suitable length by scalpel
Section, length are less than the 1/5-2/5 of coverslip length, dip in water with point of a knife, will stick up wax band, be placed on and be allowed on the water surface of glass slide
Open and flat rapidly, adjustment wax band position is allowed to marshalling.Two pieces of paper on Zhan Wentai other end pad immediately will after the flattening of wax band
Water on its glass slide blots only, then sets on warm platform；To water evaporating completely, mark is engraved in slice one end with masonry pen, is put into
Toasting in 45 DEG C of baking ovens can just carry out in next step after at least placing 1 day.
(8) dewax: piece that will be dyed is sequentially placed into 3 bottles of TO type biology clarifiers, every grade 3 minutes, slough paraffin.
(9) it dyes: piece after dewaxing is sequentially placed into+1/2 volume absolute alcohol of 1/2 volume TO type biology clarifier
Mixed liquor, absolute alcohol, absolute alcohol, volumetric concentration are 95% alcohol, volumetric concentration is 85% alcohol, volumetric concentration 70%
Rehydration in alcohol, every grade 3 seconds.It is color separation 10 seconds in 50% alcohol that piece, which 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 piece is sequentially placed into volume
Concentration is 70% alcohol, volumetric concentration is to be further processed in 85% alcohol, every grade 7 seconds.Piece is finally put into volumetric concentration
To dye in 1% fast green solution 18 seconds, the fast green solution that wherein volumetric concentration is 1% is the alcohol for being 95% with volumetric concentration
Solution is formulated for solvent.After the completion of dyeing, piece is put into absolute alcohol, rinses the coloring agent on piece, this grade is stopped
It stays 10 seconds；Piece is put into absolute alcohol again, is stopped 10 seconds；Piece is put into 1/2 volume TO type biology clarifier+1/ later
In the mixed liquor of 2 volume absolute alcohols, stop 3 seconds；Finally piece is put into TO type biology clarifier, is stopped 5 minutes, this step
Suddenly in triplicate.
(10) mounting: piece is lain on paper, wipes the remaining TO type biology clarifier in lower part, is dripped 2-3 above and is added dropwise
It puts on airs natural gum, coverslip dries on alcolhol burner, under carefully covering from side, avoids the occurrence of bubble, be put into 45 DEG C of baking ovens, dries
It can be taken off, post label, infuse the information such as name title material, collecting location, time on label.
Embodiment 2:
(1) fixed: fresh meadowrueleaf corydalis root stem adopt it is lower after with tap water wash down surface floating dust, blot blade surface moisture.So
Vertical root middle arteries are crosscutting afterwards, and every is about 5mm, wide 3mm, are put into the FAA fixer that volumetric concentration is 70% alcohol
Fix 24 hours, FAA fixer weight alcohol: glacial acetic acid: formaldehyde volume ratio is 90:5:5.
(2) it is dehydrated: material being taken out from FAA fixer, filter paper blots extra FAA fixer, is put into rubber stopper
In flat bottom glass pipe, it is in 80% alcohol that volumetric concentration, which is added, and volumetric concentration is that the additional amount of 80% alcohol is material volume
It 20 times or more, vacuum suction 15 minutes, stops 55 minutes；Pouring out volumetric concentration is 80% alcohol, and it is 85% that volumetric concentration, which is added,
Alcohol vacuum suction 10 minutes, is placed 60 minutes；Pouring out volumetric concentration is 85% alcohol, and addition volumetric concentration is 90% alcohol,
It vacuum suction 12 minutes, places 45 minutes；Pouring out volumetric concentration is 90% alcohol, and addition volumetric concentration is 95% alcohol, vacuum
Pumping 15 minutes is placed 45 minutes；Pouring out volumetric concentration is 95% alcohol, and 100% alcohol is added, vacuum suction 10 minutes, puts
It sets 45 minutes；100% alcohol is replaced, vacuum suction 15 minutes, is placed 30 minutes.
(4) transparent: to pour out absolute alcohol, before transparent, it is 0.8% that dewatered meadowrueleaf corydalis root stem, which is put in volumetric concentration,
Sarranine solution disseminate 12h, wherein volumetric concentration be 0.8% sarranine solution be with+1/3 volume TO of 2/3 volume absolute alcohol
The mixed liquor of type biology clarifier is formulated for solvent；Sarranine solution is poured out ,+1/2 volume TO of 1/2 volume absolute alcohol is added
The mixed liquor of type biology clarifier vacuum suction 10 minutes, is placed 90 minutes；Pour out+1/2 volume TO of 1/2 volume absolute alcohol
The mixed liquor of type biology clarifier, is added the mixed liquor of+2/3 volume TO type biology clarifier of 1/3 volume absolute alcohol, and vacuum is taken out
It gas 10 minutes, places 60 minutes；The mixed liquor of+2/3 volume TO type biology clarifier of 1/3 volume absolute alcohol is poured out, is added pure
TO type biology clarifier vacuum suction 10 minutes, is placed 120 minutes；TO type biology clarifier is poured out, changes a TO type biology thoroughly
It bright dose, vacuum suction 10 minutes, places 60 minutes.
(5) waxdip: the small wax stone of solid is added into the glass tube of muck scutellaria for first time waxdip, and additional amount is raw for TO type
The 1/3 of object clarifier volume, is stoppered bottle stopper, is put into 45 DEG C of insulating boxs, and same amount of wax stone is added in row again after wax thawing,
This step totally 3 times, until wax no longer melts, last time plus wax open bottle stopper, allow TO clarifier to volatilize as far as possible, this mistake
Journey needs 2 days.Second of waxdip pours into stem in crucible together with wax, then crucible is moved in 60 DEG C of insulating boxs, solidification
Wax will be poured out after all dissolving together with stem, TO clarifier, and the paraffin that molten point is 56 DEG C has been melted in change, in 60 DEG C of insulating boxs
Middle placement 4h；Then it changes to again and has melted the paraffin that molten point is 58 DEG C, can be embedded after placing 2h in 60 DEG C of insulating boxs.
(6) it embeds: taking out the crucible for filling meadowrueleaf corydalis root stem from insulating box, Label clip is gone out to be placed on embedded box on one side, bottom
After wax slightly coagulates, wax is coagulated on dissecting needle slightly hot carton surface of scratching on alcolhol burner, and the material in crucible is dialled in embedded box immediately,
And put material according to slice direction well, stand cooling.It is put into basin after surface solidification, is allowed to sink under water 30 minutes
Afterwards, embedded box is taken out, room temperature is dried, is in store for.
(7) it is sliced and exhibition piece: the fixation of wax stone and finishing: wax stone being repaired by material size and cut direction to appropriate big
It is small.Sticky wax block: the small wax stone fixed is adhered on wooden unit.It is heated on alcolhol burner with scalpel, then contacts wax stone bottom
Portion softens bottom wax, is sticked on wooden unit rapidly.The position that wax stone is connect with wooden unit should drip a little dewaxings, reinforce bonding
Fastness.The wooden unit for wax stone of adhering is fixed on slicer, required thickness (6-12 μm) is mixed up, is sliced, is put in order
It sets in ready big carton or on cardboard.The bonding die agent of minute quantity gelatin is applied on glass slide with little finger.Add on slide
Glass slide is placed on the warm platform of 50 ± (1-2) DEG C of exhibition by enough distilled water.Wax band is divided into the small of suitable length by scalpel
Section, length are less than the 1/5-2/5 of coverslip length, dip in water with point of a knife, will stick up wax band, be placed on and be allowed on the water surface of glass slide
Open and flat rapidly, adjustment wax band position is allowed to marshalling.Two pieces of paper on Zhan Wentai other end pad immediately will after the flattening of wax band
Water on its glass slide blots only, then sets on warm platform；To water evaporating completely, mark is engraved in slice one end with masonry pen, is put into
Toasting in 45 DEG C of baking ovens can just carry out in next step after at least placing 2 days.
(8) dewax: piece that will be dyed is sequentially placed into 3 bottles of TO type biology clarifiers, every grade 5 minutes, slough paraffin.
(9) it dyes: piece after dewaxing is sequentially placed into+1/2 volume absolute alcohol of 1/2 volume TO type biology clarifier
Mixed liquor, absolute alcohol, absolute alcohol, volumetric concentration are 95% alcohol, volumetric concentration is 85% alcohol, volumetric concentration 70%
Rehydration in alcohol, every grade 5 seconds.It is color separation 3 seconds in 50% alcohol that piece, which 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 piece is sequentially placed into volumetric concentration
Be to be further processed in 85% alcohol for 70% alcohol, volumetric concentration, every grade 10 seconds.Piece, which is finally put into volumetric concentration, is
It is dyed in 1.2% fast green solution 10 seconds, the fast green solution that wherein volumetric concentration is 1.2% is the wine for being 95% with volumetric concentration
Smart solution is formulated for solvent.After the completion of dyeing, piece is put into absolute alcohol, rinses the coloring agent on piece, this grade
It stops 8 seconds；Piece is put into absolute alcohol again, is stopped 3 seconds；Piece is put into 1/2 volume TO type biology clarifier+1/ later
In the mixed liquor of 2 volume absolute alcohols, stop 10 seconds；Finally piece is put into TO type biology clarifier, is stopped 10 minutes, this
Step is in triplicate.
(10) mounting: piece is lain on paper, wipes the remaining TO type biology clarifier in lower part, is dripped 2-3 above and is added dropwise
It puts on airs natural gum, coverslip dries on alcolhol burner, under carefully covering from side, avoids the occurrence of bubble, be put into 45 DEG C of baking ovens, dries
It can be taken off, post label, infuse the information such as name title material, collecting location, time on label.
Embodiment 3:
(1) fixed: fresh meadowrueleaf corydalis root stem adopt it is lower after with tap water wash down surface floating dust, blot blade surface moisture.Then
Vertical root middle arteries are crosscutting, and every is about 10mm, wide 5mm, are put into solid in the FAA fixer that volumetric concentration is 70% alcohol
24 hours fixed, FAA fixer weight alcohol: glacial acetic acid: formaldehyde volume ratio is 90:5:5.
(2) it is dehydrated: material being taken out from FAA fixer, filter paper blots extra FAA fixer, is put into rubber stopper
In flat bottom glass pipe, it is in 80% alcohol that volumetric concentration, which is added, and volumetric concentration is that the additional amount of 80% alcohol is material volume
It 20 times or more, vacuum suction 10 minutes, stops 60 minutes；Pouring out volumetric concentration is 80% alcohol, and it is 85% that volumetric concentration, which is added,
Alcohol vacuum suction 10 minutes, is placed 50 minutes；Pouring out volumetric concentration is 85% alcohol, and addition volumetric concentration is 90% alcohol,
It vacuum suction 15 minutes, places 55 minutes；Pouring out volumetric concentration is 90% alcohol, and addition volumetric concentration is 95% alcohol, vacuum
Pumping 15 minutes is placed 55 minutes；Pouring out volumetric concentration is 95% alcohol, and 100% alcohol is added, vacuum suction 15 minutes, puts
It sets 40 minutes；100% alcohol is replaced, vacuum suction 10 minutes, is placed 40 minutes.
(4) transparent: to pour out absolute alcohol, before transparent, dewatered meadowrueleaf corydalis root stem, which is put in volumetric concentration, is
0.8% sarranine solution disseminates 12h, and the sarranine solution that wherein volumetric concentration is 0.8% is with+1/3 body of 2/3 volume absolute alcohol
The mixed liquor of product TO type biology clarifier is formulated for solvent；Sarranine solution is poured out ,+1/2 body of 1/2 volume absolute alcohol is added
The mixed liquor of product TO type biology clarifier vacuum suction 10 minutes, is placed 120 minutes；Pour out+1/2 body of 1/2 volume absolute alcohol
The mixed liquor of product TO type biology clarifier, is added the mixed liquor of+2/3 volume TO type biology clarifier of 1/3 volume absolute alcohol, very
Empty pump gas 15 minutes, place 120 clocks；The mixed liquor of+2/3 volume TO type biology clarifier of 1/3 volume absolute alcohol is poured out, is added
Pure TO type biology clarifier vacuum suction 15 minutes, is placed 60 minutes；TO type biology clarifier is poured out, a TO type biology is changed
Clarifier vacuum suction 15 minutes, is placed 90 minutes.
(5) waxdip: the small wax stone of solid is added into the glass tube of muck scutellaria for first time waxdip, and additional amount is raw for TO type
The 1/3 of object clarifier volume, is stoppered bottle stopper, is put into 45 DEG C of insulating boxs, and same amount of wax stone is added in row again after wax thawing,
This step totally 4 times, until wax no longer melts, last time plus wax open bottle stopper, allow TO clarifier to volatilize as far as possible, this mistake
Journey needs 2 days.Second of waxdip pours into stem in crucible together with wax, then crucible is moved in 60 DEG C of insulating boxs, solidification
Wax will be poured out after all dissolving together with stem, TO clarifier, and the paraffin that molten point is 56 DEG C has been melted in change, in 60 DEG C of insulating boxs
Middle placement 4h；Then it changes to again and has melted the paraffin that molten point is 58 DEG C, can be embedded after placing 2h in 60 DEG C of insulating boxs.
(6) it embeds: taking out the crucible for filling meadowrueleaf corydalis root stem from insulating box, Label clip is gone out to be placed on embedded box on one side, bottom
After wax slightly coagulates, wax is coagulated on dissecting needle slightly hot carton surface of scratching on alcolhol burner, and the material in crucible is dialled in embedded box immediately,
And put material according to slice direction well, stand cooling.It is put into basin after surface solidification, is allowed to sink under water 30 minutes
Afterwards, embedded box is taken out, room temperature is dried, is in store for.
(7) it is sliced and exhibition piece: the fixation of wax stone and finishing: wax stone being repaired by material size and cut direction to appropriate big
It is small.Sticky wax block: the small wax stone fixed is adhered on wooden unit.It is heated on alcolhol burner with scalpel, then contacts wax stone bottom
Portion softens bottom wax, is sticked on wooden unit rapidly.The position that wax stone is connect with wooden unit should drip a little dewaxings, reinforce bonding
Fastness.The wooden unit for wax stone of adhering is fixed on slicer, required thickness (6-12 μm) is mixed up, is sliced, is put in order
It sets in ready big carton or on cardboard.The bonding die agent of minute quantity gelatin is applied on glass slide with little finger.Add on slide
Glass slide is placed on the warm platform of 50 ± (1-2) DEG C of exhibition by enough distilled water.Wax band is divided into the small of suitable length by scalpel
Section, length are less than the 1/5-2/5 of coverslip length, dip in water with point of a knife, will stick up wax band, be placed on and be allowed on the water surface of glass slide
Open and flat rapidly, adjustment wax band position is allowed to marshalling.Two pieces of paper on Zhan Wentai other end pad immediately will after the flattening of wax band
Water on its glass slide blots only, then sets on warm platform；To water evaporating completely, mark is engraved in slice one end with masonry pen, is put into
Toasting in 45 DEG C of baking ovens can just carry out in next step after at least placing 1 day.
(8) dewax: piece that will be dyed is sequentially placed into 3 bottles of TO type biology clarifiers, every grade 2-5 minutes, slough stone
Wax.
(9) it dyes: piece after dewaxing is sequentially placed into+1/2 volume absolute alcohol of 1/2 volume TO type biology clarifier
Mixed liquor, absolute alcohol, absolute alcohol, volumetric concentration are 95% alcohol, volumetric concentration is 85% alcohol, volumetric concentration 70%
Rehydration in alcohol, every grade 4 seconds.It is color separation 6 seconds in 50% alcohol that piece, which 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 piece is sequentially placed into volumetric concentration
Be to be further processed in 85% alcohol for 70% alcohol, volumetric concentration, every grade 3 seconds.Piece, which is finally put into volumetric concentration, is
It is dyed in 0.8% fast green solution 20 seconds, the fast green solution that wherein volumetric concentration is 0.8% is the wine for being 95% with volumetric concentration
Smart solution is formulated for solvent.After the completion of dyeing, piece is put into absolute alcohol, rinses the coloring agent on piece, this grade
It stops 3 seconds；Piece is put into absolute alcohol again, is stopped 7 seconds；Piece is put into 1/2 volume TO type biology clarifier+1/ later
In the mixed liquor of 2 volume absolute alcohols, stop 7 seconds；Finally piece is put into TO type biology clarifier, is stopped 6 minutes, this step
Suddenly in triplicate.
(10) mounting: piece is lain on paper, wipes the remaining TO type biology clarifier in lower part, is dripped 2-3 above and is added dropwise
It puts on airs natural gum, coverslip dries on alcolhol burner, under carefully covering from side, avoids the occurrence of bubble, be put into 45 DEG C of baking ovens, dries
It can be taken off, post label, infuse the information such as name title material, collecting location, time 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 carries out ultrasonic treatment 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 scutellaria carries out ultrasonic treatment 45 seconds with the ultrasonic wave that supersonic frequency is 31kHz, after dyeing the 7th hour, to putting
There is the sarranine solution of meadowrueleaf corydalis root stem to carry out ultrasonic treatment 50 seconds with the ultrasonic wave that supersonic frequency is 25kHz, continues dip dyeing to terminating.
Before the dyeing of fast green solution, being separately added into volume ratio in 70% alcohol and 85% alcohol for further processing is
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 carries out ultrasonic treatment 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 scutellaria carries out ultrasonic treatment 50 seconds with the ultrasonic wave that supersonic frequency is 30kHz, after dyeing the 7th hour, to putting
There is the sarranine solution of meadowrueleaf corydalis root stem to carry out ultrasonic treatment 30 seconds with the ultrasonic wave that supersonic frequency is 27kHz.
Before the dyeing of fast green solution, being separately added into volume ratio in 70% alcohol and 85% alcohol for further processing is
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 carries out ultrasonic treatment 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 scutellaria carries out ultrasonic treatment 60 seconds with the ultrasonic wave that supersonic frequency is 28kHz, after dyeing the 7th hour, to putting
There is the sarranine solution of meadowrueleaf corydalis root stem to carry out ultrasonic treatment 45 seconds with the ultrasonic wave that supersonic frequency is 26kHz.
Before the dyeing of fast green solution, being separately added into volume ratio in 70% alcohol and 85% alcohol for further processing is
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, applicant use conventional paraffin section method make the paraffin section of stem as
Three, is put in Lycra biomicroscope by control group, then stem's paraffin section made from Example 1 and embodiment 5 respectively
DM2000 observation is taken pictures, and respectively obtains Fig. 1, Fig. 2 and Fig. 3, as seen from the figure, the paraffin section institutional framework of Fig. 3 is clear
Clearly, it can be used for identifying 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 identifying medicinal material
The true and false.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details and legend shown and described herein.
Claims (3)
1. a kind of paraffin section method of meadowrueleaf corydalis root stem, including sample is fixed, is dehydrated, is transparent, waxdip, embedding, slice and exhibition piece,
Dewaxing, dyeing and mounting, which is characterized in that
Before fixation, meadowrueleaf corydalis root stem surface floating dust is removed, its crosscutting vertical middle arteries are long 5-10mm, the stem section of wide 3-5mm, then general
Stem section is put into FAA fixer and fixes, and FAA fixer is 70% alcohol: glacial acetic acid by volumetric concentration: formaldehyde volume ratio is 90:
5:5 is formulated；
Meadowrueleaf corydalis root stem after fixation successively passes through to volumetric concentration is 80% alcohol, volumetric concentration is 85% alcohol, volumetric concentration
It is that 95% alcohol is dehydrated step by step for 90% alcohol, volumetric concentration, each concentration gradient is dehydrated 45-60 minutes, then is put in 100% wine
It is dehydrated in essence twice, it is 30-45 minutes each；The alcoholic solution of each gradient concentration in meadowrueleaf corydalis root stem dehydration simultaneously into
Row vacuum suction 10-15 minutes；
Before transparent step, dewatered meadowrueleaf corydalis root stem is put in the sarranine solution that volumetric concentration is 0.8-1.2% and disseminates 8-
12h；The volumetric concentration is that the sarranine solution of 0.8-1.2% is transparent with+1/3 volume TO type biology of 2/3 volume absolute alcohol
The mixed liquor of agent is formulated for solvent, and it is 95% that the fast green solution that the volumetric concentration is 0.8-1.2%, which is with volumetric concentration,
Alcoholic solution be formulated for solvent；
After dewaxing, the slide for being loaded with meadowrueleaf corydalis root root is subjected to rehydration, the slide for being loaded with meadowrueleaf corydalis root stem is sequentially placed into 1/2 body
Product+1/2 volume absolute alcohol mixed liquor of type biology clarifier, absolute alcohol, absolute alcohol, volumetric concentration are 95% alcohol, body
Product concentration is 85% alcohol, volumetric concentration is rehydration in 70% alcohol, every grade 3-5 seconds；
Place into color separation in 50% alcohol after rehydration, 3-10 seconds, while volume ratio is added into the alcoholic solution for color separation and is
The acetic acid of 1/ (800-1000)；It is sequentially placed into that volumetric concentration is 70% alcohol, volumetric concentration is in 85% alcohol into one again later
Step processing, every grade 3-10 seconds；It is finally putting into the fast green solution that volumetric concentration is 0.8-1.2% and dyes 10-20 seconds；
In meadowrueleaf corydalis root stem sarranine dyeing course, the 0.8-1.2% sarranine solution for being placed with meadowrueleaf corydalis root stem is used after dyeing the 1st hour
Supersonic frequency is that the ultrasonic wave of 32-35kHz carries out ultrasonic treatment 60-75 second, after dyeing the 3rd hour, to being placed with meadowrueleaf corydalis root stem
0.8-1.2% sarranine solution carries out ultrasonic treatment 45-60 seconds with the ultrasonic wave that supersonic frequency is 28-31kHz, dyes the 7th hour
Afterwards, the 0.8-1.2% sarranine solution for being placed with meadowrueleaf corydalis root stem is ultrasonically treated with the ultrasonic wave that supersonic frequency is 25-27kHz
30-50 seconds；
The lemon that volume ratio is 1/ (1000-1200) is separately added into 70% alcohol and 85% alcohol for further processing
The mixed solution of acid, picric acid and oxalic acid, wherein citric acid in mixed solution: picric acid: the volume ratio of oxalic acid is 1:2:1.
2. the paraffin section method of meadowrueleaf corydalis root stem as described in claim 1, which is characterized in that will be loaded with after fast green solution dyeing
The slide of meadowrueleaf corydalis root stem, which is put in be put into absolute alcohol, washes away coloring agent, this grade stops 3-10 seconds；Then it is sequentially placed into no watery wine
Essence, in+1/2 absolute alcohol mixed liquor of 1/2TO type biology clarifier, every grade stop 3-10 seconds；Meadowrueleaf corydalis root stem will be finally loaded with
Slide is put into TO type biology clarifier and stops 5-10 minutes, this step is in triplicate.
3. the paraffin section method of meadowrueleaf corydalis root stem as described in claim 1, which is characterized in that the stem after the dyeing of sarranine solution
Successively with+1/2 volume TO type biology clarifier mixed liquor of 1/2 volume absolute alcohol and+2/3 volume of 1/3 volume absolute alcohol
TO type biology clarifier mixed liquor carries out transparent processing, and every grade of clearing time is 60-120 minutes；It is put in pure TO type biology again
It impregnates 2 times in clarifier, impregnates 60-120 minutes every time.
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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 |
| CN113933136B (en) * | 2021-10-15 | 2023-06-23 | 华北理工大学 | Dendrite corrosion reagent of medical zinc-based alloy without Al, preparation and use method thereof |
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| CN1068889A (en) * | 1992-07-11 | 1993-02-10 | 南京大学 | The method and apparatus of quick preparing human tissue pathological section |
| CN102967493A (en) * | 2012-10-27 | 2013-03-13 | 山西农业大学 | Rapid paraffin sectioning method for plant tissue |
| CN103940658A (en) * | 2014-04-10 | 2014-07-23 | 青岛大学医学院附属医院 | Method for manufacturing paraffin-embedded tissue cell specimen |
| CN105973673A (en) * | 2016-06-30 | 2016-09-28 | 中国林业科学研究院热带林业研究所 | Paraffin sectioning method for eucalyptus tissue |
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| CN1068889A (en) * | 1992-07-11 | 1993-02-10 | 南京大学 | The method and apparatus of quick preparing human tissue pathological section |
| CN102967493A (en) * | 2012-10-27 | 2013-03-13 | 山西农业大学 | Rapid paraffin sectioning method for plant tissue |
| CN103940658A (en) * | 2014-04-10 | 2014-07-23 | 青岛大学医学院附属医院 | Method for manufacturing paraffin-embedded tissue cell specimen |
| CN105973673A (en) * | 2016-06-30 | 2016-09-28 | 中国林业科学研究院热带林业研究所 | Paraffin sectioning method for eucalyptus tissue |
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