CN110967242A - Method for analyzing lipid plaque content by fixing rabbit aorta - Google Patents
Method for analyzing lipid plaque content by fixing rabbit aorta Download PDFInfo
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- CN110967242A CN110967242A CN201911322530.7A CN201911322530A CN110967242A CN 110967242 A CN110967242 A CN 110967242A CN 201911322530 A CN201911322530 A CN 201911322530A CN 110967242 A CN110967242 A CN 110967242A
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- artery
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- lipid plaque
- sudan
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- 238000000034 method Methods 0.000 title claims abstract description 26
- 150000002632 lipids Chemical class 0.000 title claims abstract description 24
- 241000283973 Oryctolagus cuniculus Species 0.000 title claims abstract description 22
- 210000000709 aorta Anatomy 0.000 title claims abstract description 22
- 210000001367 artery Anatomy 0.000 claims abstract description 44
- 229930040373 Paraformaldehyde Natural products 0.000 claims abstract description 9
- 238000004043 dyeing Methods 0.000 claims abstract description 9
- 229920002866 paraformaldehyde Polymers 0.000 claims abstract description 9
- 238000002791 soaking Methods 0.000 claims abstract description 7
- 239000002390 adhesive tape Substances 0.000 claims abstract description 5
- 238000007493 shaping process Methods 0.000 claims abstract description 5
- 238000002360 preparation method Methods 0.000 claims abstract description 4
- 239000002994 raw material Substances 0.000 claims abstract description 4
- 239000003086 colorant Substances 0.000 claims description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- 210000002376 aorta thoracic Anatomy 0.000 claims description 6
- 238000013499 data model Methods 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims description 3
- 238000000576 coating method Methods 0.000 claims description 3
- 238000005520 cutting process Methods 0.000 claims description 3
- 210000003692 ilium Anatomy 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 230000010412 perfusion Effects 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 abstract description 3
- 239000007788 liquid Substances 0.000 abstract description 2
- RCTGMCJBQGBLKT-UHFFFAOYSA-N Sudan IV Chemical compound CC1=CC=CC=C1N=NC(C=C1C)=CC=C1N=NC1=C(O)C=CC2=CC=CC=C12 RCTGMCJBQGBLKT-UHFFFAOYSA-N 0.000 abstract 1
- 239000002344 surface layer Substances 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- 238000011160 research Methods 0.000 description 3
- 230000003100 immobilizing effect Effects 0.000 description 2
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—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
-
- 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
- G01N2001/305—Fixative compositions
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- 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)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the technical field of biological experiments, in particular to a method for analyzing lipid plaque content by fixing rabbit aorta, which comprises the following steps: A. selecting raw materials; B. selecting an artery; C. and (3) artery shaping: washing an artery sample, and soaking the artery sample in a prepared paraformaldehyde fixing solution for 24 hours; D. fat marking: taking out the shaped artery sample, dissecting from top to bottom, and dyeing the lipid plaque by a Sudan IV dyeing method; E. preparation of a sample plate: sticking the black double-sided adhesive tape on the transparent sample plate, and sticking the dyed artery sample on the surface layer of the black double-sided adhesive tape; F. and (4) scanning and analyzing. According to the invention, through a proper fixing mode, the artery can be rapidly flattened so as to obtain a clear lipid plaque map of the inner wall of the artery, then the fixing and sudan IV stain fat marking are sequentially completed by adopting paraformaldehyde fixing liquid, the artery curling or the distinguishing is not obvious in the pasting mode, and finally the prepared sample plate can rapidly and comprehensively scan the artery, so that the fixation of the rabbit aorta is simplified and the analysis efficiency of the lipid content is improved.
Description
Technical Field
The invention relates to the technical field of biological experiments, in particular to a method for analyzing lipid plaque content by fixing rabbit aorta.
Background
The rabbit is a commonly used research object in biological experiments, for example, the research on the content of lipid plaques in the aorta of the rabbit can help technicians to better understand the physiological structure of similar organisms, thereby facilitating other related researches at a later stage.
However, because no good method for fixing the rabbit artery exists at present, the rabbit artery cannot be well flattened and fixedly scanned, and the problems that one-time scanning is insufficient, multiple times of scanning wastes time, scanning images are not visual and the like are often caused.
Disclosure of Invention
The invention aims to provide a method for analyzing lipid plaque content by fixing rabbit aorta, which has the advantages of convenience in fixation and intuition in detection and solves the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme: a method for immobilizing rabbit aorta for analyzing lipid plaque content, comprising the following steps:
A. selecting raw materials: taking out the complete branches of the neck and the ilium of the rabbit containing the artery without perfusion, and stripping the redundant fat at the periphery;
B. selecting artery nodes: stopping from the upper edge of the aortic arch to the aortic arch root, cutting the lower edge of the aorta to the iliac part of the aorta, and stretching and flattening to obtain an artery sample meeting the requirement;
C. and (3) artery shaping: soaking an artery sample in a prepared paraformaldehyde fixing solution for a period of time;
D. fat marking: taking out the shaped artery sample, and dyeing the lipid plaque by adopting a Sudan IV dyeing method;
E. preparation of a sample plate: adhering the stained artery sample to the surface of the transparent plate to obtain a sample plate;
F. scanning and analyzing: and (4) reversely buckling the sample plate on a scanner, scanning to obtain a fixed artery data model with plaque, and analyzing the artery data model.
Preferably, the concentration ratio of the paraformaldehyde fixing solution in the step C is 4%.
Preferably, the soaking time in the step C is 30min-60 min.
Preferably, the specific content of step D is as follows,
weighing 0.1g of Sudan IV dry powder (dark brown), dissolving in 50ml of acetone, adding 50ml of 70% alcohol by volume, fully mixing to obtain a Sudan IV coloring agent, uniformly dripping the Sudan IV coloring agent on an artery sample by using a dropper, uniformly coating, and standing for a moment to enable all lipid plaques to be red plaques.
Preferably, an ultrathin high-transparency double-sided adhesive tape is used as the adhering tool in the step E.
Preferably, the transparent plate in the step E is an acrylic plate.
Compared with the prior art, the invention has the following beneficial effects:
the method comprises the steps of selecting and flattening an artery sample with a proper length, then finishing shaping and fat marking by sequentially adopting paraformaldehyde fixing liquid and Sudan IV coloring agent, avoiding the artery from curling or being indistinct, and finally making a sample plate to facilitate rapid and comprehensive scanning, thereby greatly improving the efficiency of fixing the rabbit aorta and analyzing the lipid content.
Drawings
FIG. 1 is a process flow diagram of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Please refer to fig. 1, in which:
a method for immobilizing rabbit aorta for analyzing lipid plaque content, comprising the following steps:
A. selecting raw materials: taking out the complete branches of the neck and the ilium of the rabbit containing the artery without perfusion, and stripping the redundant fat at the periphery;
B. selecting artery nodes: stopping from the upper edge of the aortic arch to the aortic arch root, cutting the lower edge of the aorta to the iliac part of the aorta, and stretching and flattening to obtain an artery sample meeting the requirement;
C. and (3) artery shaping: soaking an artery sample in a prepared paraformaldehyde fixing solution for a period of time;
D. fat marking: taking out the shaped artery sample, and dyeing the lipid plaque by adopting a Sudan IV dyeing method;
E. preparation of a sample plate: adhering the stained artery sample to the surface of the transparent plate to obtain a sample plate;
F. scanning and analyzing: and (4) reversely buckling the sample plate on a scanner, scanning to obtain a fixed artery data model with plaque, and analyzing the artery data model.
And C, the concentration ratio of the paraformaldehyde fixing solution in the step C is 4%.
And the soaking time in the step C is 30-60 min.
The specific content of the step D is as follows,
weighing 0.1g of Sudan IV dry powder (dark brown), dissolving in 50ml of acetone, adding 50ml of 70% alcohol by volume, fully mixing to obtain a Sudan IV coloring agent, uniformly dripping the Sudan IV coloring agent on an artery sample by using a dropper, uniformly coating, and standing for a moment to enable all lipid plaques to be red plaques.
And in the step E, an ultrathin high-transparency double-sided adhesive tape is used as a pasting tool.
And E, adopting an acrylic plate as the transparent plate.
Control group one:
the content of the comparison group is basically the same as that of the embodiment, and the same parts are not repeated, except that: and D, the step C is cancelled, and the dyeing link in the step D is directly carried out.
Control group two:
the content of the comparison group is basically the same as that of the embodiment, and the same parts are not repeated, except that: and E, eliminating the step E, and scanning by a conventional clamping mode.
Table one:
| examples | Control group one | Control group two | |
| Degree of scan integrity | Height of | Is low in | Is higher than |
| Number of scans | 1 | 1 | ＞1 |
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (6)
1. A method for analyzing lipid plaque content by fixing rabbit aorta is characterized in that: the method comprises the following steps:
A. selecting raw materials: taking out the complete branches of the neck and the ilium of the rabbit containing the artery without perfusion, and stripping the redundant fat at the periphery;
B. selecting artery nodes: stopping from the upper edge of the aortic arch to the aortic arch root, cutting the lower edge of the aorta to the iliac part of the aorta, and stretching and flattening to obtain an artery sample meeting the requirement;
C. and (3) artery shaping: soaking an artery sample in a prepared paraformaldehyde fixing solution for a period of time;
D. fat marking: taking out the shaped artery sample, and dyeing the lipid plaque by adopting a Sudan IV dyeing method;
E. preparation of a sample plate: adhering the stained artery sample to the surface of the transparent plate to obtain a sample plate;
F. scanning and analyzing: and (4) reversely buckling the sample plate on a scanner, scanning to obtain a fixed artery data model with plaque, and analyzing the artery data model.
2. The method of claim 1, wherein the rabbit aorta is fixed for analysis of lipid plaque content, and the method comprises: and C, the concentration ratio of the paraformaldehyde fixing solution in the step C is 4%.
3. The method of claim 1, wherein the rabbit aorta is fixed for analysis of lipid plaque content, and the method comprises: and the soaking time in the step C is 30-60 min.
4. The method of claim 1, wherein the rabbit aorta is fixed for analysis of lipid plaque content, and the method comprises: the specific content of the step D is as follows,
weighing 0.1g of Sudan IV dry powder (dark brown), dissolving in 50ml of acetone, adding 50ml of 70% alcohol by volume, fully mixing to obtain a Sudan IV coloring agent, uniformly dripping the Sudan IV coloring agent on an artery sample by using a dropper, uniformly coating, and standing for a moment to enable all lipid plaques to be red plaques.
5. The method of claim 1, wherein the rabbit aorta is fixed for analysis of lipid plaque content, and the method comprises: and in the step E, an ultrathin high-transparency double-sided adhesive tape is used as a pasting tool.
6. The method of claim 1, wherein the rabbit aorta is fixed for analysis of lipid plaque content, and the method comprises: and E, adopting an acrylic plate as the transparent plate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911322530.7A CN110967242A (en) | 2019-12-20 | 2019-12-20 | Method for analyzing lipid plaque content by fixing rabbit aorta |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911322530.7A CN110967242A (en) | 2019-12-20 | 2019-12-20 | Method for analyzing lipid plaque content by fixing rabbit aorta |
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| CN110967242A true CN110967242A (en) | 2020-04-07 |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104490885A (en) * | 2014-11-26 | 2015-04-08 | 湖南中医药大学 | Method for establishing domestic rabbit atherosclerosis model |
| CN105012041A (en) * | 2015-07-13 | 2015-11-04 | 高不郎 | Mold for manufacturing aneurysm model and method |
| CN105816549A (en) * | 2016-04-26 | 2016-08-03 | 南方医科大学 | Medicine for preventing and treating atherosclerotic plaque and preparation method thereof |
| CN108514462A (en) * | 2018-06-11 | 2018-09-11 | 新乡医学院 | A kind of mouse perfusion fixing device and its perfusion fixing means |
| CN108753837A (en) * | 2018-06-15 | 2018-11-06 | 扬州大学 | The construction method and sgRNA of a kind of hyperlipidemia or rabbit model |
| CN109187147A (en) * | 2018-08-27 | 2019-01-11 | 广州医科大学附属第医院 | A kind of dyeing of arterial wall ingredient and identification method |
-
2019
- 2019-12-20 CN CN201911322530.7A patent/CN110967242A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104490885A (en) * | 2014-11-26 | 2015-04-08 | 湖南中医药大学 | Method for establishing domestic rabbit atherosclerosis model |
| CN105012041A (en) * | 2015-07-13 | 2015-11-04 | 高不郎 | Mold for manufacturing aneurysm model and method |
| CN105816549A (en) * | 2016-04-26 | 2016-08-03 | 南方医科大学 | Medicine for preventing and treating atherosclerotic plaque and preparation method thereof |
| CN108514462A (en) * | 2018-06-11 | 2018-09-11 | 新乡医学院 | A kind of mouse perfusion fixing device and its perfusion fixing means |
| CN108753837A (en) * | 2018-06-15 | 2018-11-06 | 扬州大学 | The construction method and sgRNA of a kind of hyperlipidemia or rabbit model |
| CN109187147A (en) * | 2018-08-27 | 2019-01-11 | 广州医科大学附属第医院 | A kind of dyeing of arterial wall ingredient and identification method |
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