CN105651581A - Staining agent and preparation method and application thereof - Google Patents
Staining agent and preparation method and application thereof Download PDFInfo
- Publication number
- CN105651581A CN105651581A CN201610202128.5A CN201610202128A CN105651581A CN 105651581 A CN105651581 A CN 105651581A CN 201610202128 A CN201610202128 A CN 201610202128A CN 105651581 A CN105651581 A CN 105651581A
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- China
- Prior art keywords
- stain
- preparation
- picric acid
- alcoholic solution
- staining agent
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- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 238000010186 staining Methods 0.000 title abstract description 12
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 49
- 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 abstract description 35
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000000243 solution Substances 0.000 claims description 59
- 230000001476 alcoholic effect Effects 0.000 claims description 33
- 238000000034 method Methods 0.000 claims description 14
- 239000012153 distilled water Substances 0.000 claims description 13
- 238000010438 heat treatment Methods 0.000 claims description 12
- 239000000047 product Substances 0.000 claims description 9
- 229920003023 plastic Polymers 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 4
- 239000011259 mixed solution Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 abstract description 9
- 210000003484 anatomy Anatomy 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 230000005540 biological transmission Effects 0.000 abstract description 3
- 238000002224 dissection Methods 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract 4
- 241000130764 Tinea Species 0.000 description 19
- 208000002474 Tinea Diseases 0.000 description 19
- 238000002474 experimental method Methods 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 241000270959 Pelophylax nigromaculatus Species 0.000 description 3
- 210000001595 mastoid Anatomy 0.000 description 3
- 241000252212 Danio rerio Species 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 238000004040 coloring Methods 0.000 description 2
- 210000004087 cornea Anatomy 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 210000003725 endotheliocyte Anatomy 0.000 description 2
- 210000003141 lower extremity Anatomy 0.000 description 2
- 230000029052 metamorphosis Effects 0.000 description 2
- 208000003464 asthenopia Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 238000010827 pathological analysis Methods 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 210000003660 reticulum Anatomy 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000011121 vaginal smear 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the technical field of chemical staining agents, in particular to a staining agent and a preparation method and application thereof. The staining agent comprises a picric acid solution, an ethanol solution and formalin, wherein by volume, the ethanol solution accounts for 50-75% of the staining agent. The staining agent and the preparation method and application thereof have the advantages that the staining agent is excellent in staining effect, transparent in color, easy to elute in a later period and less prone to changing sample hardness; when the staining agent is applied to anatomy of biomedicine, samples can be displayed quickly, efficiently and accurately, staining agent consumption is reduced and sample integrity is also protected; the problem that the young and tender dissected samples large in water content, high in light transmission and soft are observed difficultly is solved beneficially, animal dissection structure observation effects can be improved remarkably, and labor strength of observers is reduced.
Description
Technical field
The present invention relates to chemical staining agent technical field, particularly to a kind of stain and its preparation method and application.
Background technology
In recent years, along with the development of medical science and going deep into of pathological study, the substantial amounts of anatomy experiment of human needs goes to be analyzed and detect, for for referencial use for further pathological analysis, so when we are when carrying out biomedical anatomy experiment, sample is carried out dyeing and can observe anatomical structure well.
Generally, biomedical anatomy experiment is to analyze tissue morphology, mainly through whole tissue is dyeed so that slough tissue can clearly be observed under the microscope. Conventional colouring method has: hematoxylin eosin stain method, Papanicolaou's vaginal smear technique, shore staining, Mai Geji dyeing etc., but, these methods all have subjectivity strongly, it is easily caused analysis not accurate enough, and most tissue water contents are big, light transmission strong, children is tender, quality is soft, direct perusal or stereoscope are observed and are all not easily seen its architectural feature clearly, therefore, self quality characteristic and methods for using them of stain is particularly important for Color, affects the accuracy of interpretation of result further.
Summary of the invention
The present invention solves above-mentioned technical problem, it is provided that a kind of stain and preparation method thereof and application process.
It is achieved especially by below scheme:
A kind of stain, described stain at least contains picric acid solution and alcoholic solution, and the volumetric usage accounting of alcoholic solution is 50%-75%.
The volumetric concentration of described alcoholic solution is 50%-85%.
The percent by volume of each component of described stain: picric acid solution 15-25%, alcoholic solution 50%-75%, all the other are distilled water.
Described stain is possibly together with formalin.
In described formalin, content of formaldehyde accounting is 3.5%-4%, it is preferred that, in formalin, content of formaldehyde accounting is 4%.
Its preparation method, comprises the following steps:
(1) preparation of picric acid solution: take picric acid and be placed in transparent plastic bottle, adds distilled water and is sufficiently mixed, staticly settle, take the supernatant;
(2) being mixed homogeneously at normal temperatures with alcoholic solution by picric acid solution, wherein, the volumetric concentration of alcoholic solution is 50%-85%, then adds or without formalin, shakes up, it is thus achieved that stain.
Described mix homogeneously, it is placed under the environment that temperature is 60 DEG C-70 DEG C and carries out heating in water bath 5min-10min in the completed, adds or without formalin in heating in water bath process.
Described water-bath, its after completion, then mixed solution is placed in temperature be 40 DEG C-45 DEG C heating 15s-30s, vibrate 15min-20min, cooling.
Described stain, its application in biomedicine.
Further, described stain, in application process, carries out by the following method: drips stain to product to be seen to product to be seen and is submerged, then under normal temperature environment, after will drown out the product to be seen dip-dye 20min-40min processed, it is placed in stereoscopic Microscopic observation.
Beneficial effects of the present invention
It is good that the stain of the present invention has coloring effect; color is clear and later stage easy eluting; the not feature of malleable sample hardness; it is applied in biomedical anatomy; make the sample can quickly, efficiently, show accurately; also reduce stain consumption, also protect the integrity of sample simultaneously.
Further combined with application process, solve the difficult problem that the dissection sample that water content is big, light transmission strong, children is tender, quality is soft is not easily observed, also significantly increase the observing effect of zootomy structure simultaneously, improve the situation of the easy visual fatigue of observer, reduce labor intensity.
Accompanying drawing explanation
Fig. 1: use the Tinea Ranae lips and teeth formula design sketch of stain of the present invention;
Fig. 2: do not use the Tinea Ranae lips and teeth formula design sketch of stain of the present invention.
Detailed description of the invention
Below in conjunction with specific embodiment technical scheme done further restriction, but the scope claimed is not only limited to the description made.
Embodiment 1
A kind of stain, the percent by volume of each component: picric acid solution 25%, alcoholic solution 50%, all the other are distilled water.
The volumetric concentration of described alcoholic solution is 50%.
Its preparation method, comprises the following steps:
(1) preparation of picric acid solution: take picric acid and be placed in transparent plastic bottle, adds distilled water and is sufficiently mixed, staticly settle, take the supernatant;
(2) being mixed homogeneously at normal temperatures with alcoholic solution by picric acid solution, wherein, the volumetric concentration of alcoholic solution is 50%, then shakes up, it is thus achieved that stain.
It is applied to prepared stain observe Tinea Ranae lips and teeth.
Embodiment 2
A kind of stain, the percent by volume of each component: picric acid solution 20%, formalin 5%, alcoholic solution 75%.
The volumetric concentration of described alcoholic solution is 85%.
Its preparation method, comprises the following steps:
(1) preparation of picric acid solution: take picric acid and be placed in transparent plastic bottle, adds distilled water and is sufficiently mixed, staticly settle, take the supernatant;
(2) being mixed homogeneously at normal temperatures with alcoholic solution by picric acid solution, wherein, the volumetric concentration of alcoholic solution is 85%, then shakes up, it is thus achieved that stain.
It is applied to prepared stain observe Tinea Ranae lips and teeth.
Embodiment 3
A kind of stain, the percent by volume of each component: picric acid solution 15%, formalin 10%, alcoholic solution 60%.
The volumetric concentration of described alcoholic solution is 65%.
Its preparation method, comprises the following steps:
(1) preparation of picric acid solution: take picric acid and be placed in transparent plastic bottle, adds distilled water and is sufficiently mixed, staticly settle, take the supernatant;
(2) being mixed homogeneously at normal temperatures with alcoholic solution by picric acid solution, wherein, the volumetric concentration of alcoholic solution is 65%, then shakes up, it is thus achieved that stain.
It is applied to prepared stain observe Tinea Ranae lips and teeth.
Embodiment 4
A kind of stain, the percent by volume of each component: picric acid solution 15%, formalin 10%, alcoholic solution 60%, all the other are distilled water.
The volumetric concentration of described alcoholic solution is 70%.
Its preparation method, comprises the following steps:
(1) preparation of picric acid solution: take picric acid and be placed in transparent plastic bottle, adds the distilled water that temperature is 80 DEG C and is sufficiently mixed, staticly settle, then take the supernatant;
(2) picric acid solution is mixed homogeneously at normal temperatures with alcoholic solution, be subsequently placed under the environment that temperature is 60 DEG C and carry out heating in water bath 10min, then be placed in microwave heating 15s under the environment that temperature is 40 DEG C, through vibration 20min, cooling, it is thus achieved that stain.
It is applied to prepared stain observe Tinea Ranae lips and teeth; Observed result shows: the upper lips and teeth the 1st of Rana nigromaculata Tinea Ranae are drained whole, becomes a line about the 2nd row's terminal, and lower lips and teeth get close to a row of horny jaw and divide right and left.
Embodiment 5
A kind of stain, the percent by volume of each component: picric acid solution 15%, formalin 10%, alcoholic solution 60%, all the other are distilled water.
The volumetric concentration of described alcoholic solution is 75%.
Its preparation method, comprises the following steps:
(1) preparation of picric acid solution: take picric acid and be placed in transparent plastic bottle, adds the distilled water that temperature is 100 DEG C and is sufficiently mixed, staticly settle, then take the supernatant;
(2) picric acid solution is mixed homogeneously at normal temperatures with the alcoholic solution that volumetric concentration is 75%, it is subsequently placed under the environment that temperature is 70 DEG C, add the formalin that content of formaldehyde accounting is 4% and carry out heating in water bath 8min, it is placed in microwave heating 30s under the environment that temperature is 45 DEG C again, through vibration 15min, cooling, it is thus achieved that stain.
It is applied to prepared stain observe Adult Zebrafish tooth; Observed result shows: Adult Zebrafish tooth is acrodont, triangular in shape, is positioned on the 5th cheek bow.
Embodiment 6
A kind of stain, the percent by volume of each component: picric acid solution 20%, formalin 5%, alcoholic solution 75%.
The volumetric concentration of described alcoholic solution is 85%.
Its preparation method, comprises the following steps:
(1) preparation of picric acid solution: take picric acid and be placed in transparent plastic bottle, adds the distilled water that temperature is 115 DEG C and is sufficiently mixed, staticly settle, then take the supernatant;
(2) it is placed under room temperature by picric acid solution with the alcoholic solution that volumetric concentration is 85% to mix homogeneously, it is subsequently placed under the environment that temperature is 65 DEG C, add the formalin that content of formaldehyde accounting is 3.5% and carry out heating in water bath 5min, it is placed in microwave heating 20s under the environment that temperature is 43 DEG C again, through vibration 18min, it is cooled to room temperature, it is thus achieved that stain.
It is applied to prepared stain observe rabbit cornea endotheliocyte; Observed result: the peplos of rabbit cornea endotheliocyte is imperfect, has cavity to be formed in endochylema, reticulum dilatation, and nuclear swelling deforms.
Test example 1
Choosing each 3 of the Yunnan frog Tinea Ranae of hind limb bud period of development, 5 toe idiophases, period of metamorphosis respectively, the Rana nigromaculata Tinea Ranae in each period is carried out living specimen observation, and records the lip of Rana nigromaculata Tinea Ranae, labial papilia and lips and teeth, specific experiment method is:
Step 1: with hind limb bud budding Yunnan frog Tinea Ranae for sample, the stain dripping embodiment 1 to sample is submerged to Tinea Ranae, then under normal temperature environment, after will drown out the product to be seen dip-dye 20min processed, being placed in stereoscopic Microscopic observation, observed result is: Yunnan frog Tinea Ranae oral area lip arc, has horny jaw, its lips and teeth formula is I: 1-1/1-1: II, lower labial papilia 2 is arranged, and one-tenth is staggered, and the mastoid process of the relatively interior row of outer row is long, middle 3-4 grain is the longest, has anapophysis.
Step 2: with the Yunnan frog Tinea Ranae of 5 toe idiophases for sample, the stain dripping embodiment 2 to sample is submerged to Tinea Ranae, then under normal temperature environment, will drown out the product to be seen processed to contaminate: after 35min, being placed in stereoscopic Microscopic observation, observed result is: Yunnan frog Tinea Ranae oral area lip arc, there is horny jaw, its lips and teeth formula is I: 1-1/1-1: II, and lower labial papilia 2 is arranged, and one-tenth is staggered, the mastoid process of the relatively interior row of outer row is long, middle 3-4 grain is the longest, has anapophysis, and lips and teeth and lips and teeth rib strengthen.
Step 3: with the Yunnan frog Tinea Ranae of period of metamorphosis for sample, the stain dripping embodiment 3 to sample is submerged to Tinea Ranae, then under normal temperature environment, after will drown out the product to be seen dip-dye 40min processed, it is placed in stereoscopic Microscopic observation, observed result is: the lips and teeth formula of Yunnan frog Tinea Ranae is I: 1-1/1-1: II, and lower labial papilia 2 is arranged, and one-tenth is staggered, the mastoid process of the relatively interior row of outer row is long, middle 3-4 grain is the longest, has anapophysis, but lip, horny jaw disappear.
Claims (10)
1. a stain, it is characterised in that described stain at least contains picric acid solution and alcoholic solution, and the volumetric usage accounting of alcoholic solution is 50%-75%.
2. stain as claimed in claim 1, it is characterised in that the volumetric concentration of described alcoholic solution is 50%-85%.
3. stain as claimed in claim 1 or 2, it is characterised in that the percent by volume of each component of described stain: picric acid solution 15-25%, alcoholic solution 50%-75%, all the other are distilled water.
4. stain as claimed in claim 1 or 2, it is characterised in that described stain is possibly together with formalin.
5. stain as claimed in claim 4, it is characterised in that in described formalin, content of formaldehyde accounting is 3.5%-4%.
6. the preparation method of the stain as described in any one of claim 1-5, it is characterised in that comprise the following steps:
(1) preparation of picric acid solution: take picric acid and be placed in transparent plastic bottle, adds distilled water and is sufficiently mixed, staticly settle, take the supernatant;
(2) being mixed homogeneously at normal temperatures with alcoholic solution by picric acid solution, wherein, the volumetric concentration of alcoholic solution is 50%-85%, then adds or without formalin, shakes up, it is thus achieved that stain.
7. the preparation method of stain as claimed in claim 6, it is characterized in that, described mix homogeneously, it is in the completed, it is placed under the environment that temperature is 60 DEG C-70 DEG C and carries out heating in water bath 5min-10min, add or without formalin in heating in water bath process.
8. the preparation method of stain as claimed in claim 6, it is characterised in that described water-bath, its after completion, then mixed solution is placed in temperature is 40 DEG C-45 DEG C heating 15s-30s, and vibrate 15min-20min, cooling.
9. the stain that prepared by the preparation method of stain as described in any one of claim 1-5 or stain as claimed in claim 6, its application in biomedicine.
10. stain application in biomedicine as claimed in claim 9, it is characterized in that, it is in application process, carry out by the following method: drip stain to product to be seen to product to be seen and be submerged, again under normal temperature environment, after will drown out the product to be seen dip-dye 20min-40min processed, it is placed in stereoscopic Microscopic observation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610202128.5A CN105651581A (en) | 2016-03-31 | 2016-03-31 | Staining agent and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610202128.5A CN105651581A (en) | 2016-03-31 | 2016-03-31 | Staining agent and preparation method and application thereof |
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| Publication Number | Publication Date |
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| CN105651581A true CN105651581A (en) | 2016-06-08 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
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| CN201610202128.5A Pending CN105651581A (en) | 2016-03-31 | 2016-03-31 | Staining agent and preparation method and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107603276A (en) * | 2017-09-14 | 2018-01-19 | 郑州乐业生物科技有限公司 | A kind of biological stain of stabilization and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130011854A1 (en) * | 2008-12-03 | 2013-01-10 | The United States Government as represented by the Dept. of Veterans Affairs | Pressure-assisted molecular recovery (pamr) of biomolecules, pressure-assisted antigen retrieval (paar), and pressure-assisted tissue histology (path) |
| CN105043841A (en) * | 2015-06-25 | 2015-11-11 | 中国人民解放军第四军医大学 | Method for staining hard tissue slices using picrosirius red and application of method |
-
2016
- 2016-03-31 CN CN201610202128.5A patent/CN105651581A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20130011854A1 (en) * | 2008-12-03 | 2013-01-10 | The United States Government as represented by the Dept. of Veterans Affairs | Pressure-assisted molecular recovery (pamr) of biomolecules, pressure-assisted antigen retrieval (paar), and pressure-assisted tissue histology (path) |
| CN105043841A (en) * | 2015-06-25 | 2015-11-11 | 中国人民解放军第四军医大学 | Method for staining hard tissue slices using picrosirius red and application of method |
Non-Patent Citations (1)
| Title |
|---|
| 丁伟 等: "《简明病理学技术》", 28 February 2014, 浙江科学技术出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107603276A (en) * | 2017-09-14 | 2018-01-19 | 郑州乐业生物科技有限公司 | A kind of biological stain of stabilization and preparation method thereof |
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Application publication date: 20160608 |