CN110967242A - A method for immobilizing rabbit aorta for analyzing lipid plaque content - Google Patents

A method for immobilizing rabbit aorta for analyzing lipid plaque content Download PDF

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Publication number
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
lipid
sample
aorta
content
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梁景岩
严坤宁
王英歌
庄文雯
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Yangzhou University
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Yangzhou University
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N2001/302Stain compositions
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N2001/305Fixative compositions

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  • Life Sciences & Earth Sciences (AREA)
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  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
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  • General Health & Medical Sciences (AREA)
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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

Method for analyzing lipid plaque content by fixing rabbit aorta
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)

1.一种固定家兔大动脉用以分析脂质斑块含量的方法,其特征在于:包括以下步骤:1. a fixed rabbit aorta is used to analyze the method for lipid plaque content, it is characterized in that: comprise the following steps: A、选取原材料:无需灌注,取出包含动脉的家兔颈部、髂部的完整分支,剥离外周多余的脂肪;A. Selection of raw materials: without perfusion, the complete branches of the neck and iliac part of the rabbit including the artery were taken out, and the excess peripheral fat was stripped; B、动脉节选:沿主动脉弓上缘至主动脉弓根停止,主动脉下缘剪至主动脉髂部,再拉伸展平,即可制得符合需要的动脉样本;B. Arterial selection: along the upper edge of the aortic arch to the root of the aortic arch, cut the lower edge of the aorta to the iliac part of the aorta, and then stretch and flatten to obtain the desired arterial sample; C、动脉定型:将动脉样本放入预配的多聚甲醛固定液中浸泡一段时间;C. Arterial stereotyping: soak the arterial samples in the pre-prepared paraformaldehyde fixative solution for a period of time; D、脂肪标记:将定型后的动脉样本取出,并采用苏丹Ⅳ染色的方法对脂质斑块进行染色;D. Fat labeling: take out the stereotyped arterial samples, and use Sudan IV staining to stain the lipid plaques; E、样板制备:将染色后的动脉样本粘贴于透明板的表面,获得样板;E. Template preparation: paste the stained arterial sample on the surface of the transparent plate to obtain a template; F、扫描分析:将样板倒扣在扫描仪上,扫描即可得到固定好的有斑块的动脉数据模型,并对其进行分析。F. Scanning analysis: put the template upside down on the scanner, scan to get the fixed plaque data model, and analyze it. 2.根据权利要求1所述的一种固定家兔大动脉用以分析脂质斑块含量的方法,其特征在于:所述步骤C中多聚甲醛固定液的浓度比为4%。2 . The method for fixing the aorta of rabbits according to claim 1 to analyze lipid plaque content, wherein the concentration ratio of the paraformaldehyde fixative in the step C is 4%. 3 . 3.根据权利要求1所述的一种固定家兔大动脉用以分析脂质斑块含量的方法,其特征在于:所述步骤C中浸泡时间为30min-60min。3. a kind of fixed rabbit aorta according to claim 1 is in order to analyze the method for lipid plaque content, it is characterized in that: in described step C, soaking time is 30min-60min. 4.根据权利要求1所述的一种固定家兔大动脉用以分析脂质斑块含量的方法,其特征在于:所述步骤D的具体内容如下,4. a kind of fixed rabbit aorta according to claim 1 is in order to analyze the method for lipid plaque content, it is characterized in that: the concrete content of described step D is as follows, 称取0.1g苏丹Ⅳ干粉(深褐色),溶于50ml丙酮中,再加入体积分数为70%的酒精50ml,充分混合后即可制得苏丹Ⅳ染色剂,然后使用滴管在动脉样本上均匀滴加苏丹Ⅳ染色剂并涂匀,静置片刻,即可使得所有脂质斑块呈现为红色斑块。Weigh 0.1g of Sudan IV dry powder (dark brown), dissolve it in 50ml of acetone, add 50ml of alcohol with a volume fraction of 70%, and mix thoroughly to obtain Sudan IV stain, and then use a dropper to spread evenly on the arterial sample Add Sudan IV dye dropwise and spread it evenly, and let it stand for a while, all lipid plaques will appear as red plaques. 5.根据权利要求1所述的一种固定家兔大动脉用以分析脂质斑块含量的方法,其特征在于:所述步骤E中采用超薄高透明双面胶作为粘贴工具。5. a kind of fixed rabbit aorta according to claim 1 is in order to analyze the method for lipid plaque content, it is characterized in that: in described step E, adopt ultra-thin high-transparency double-sided adhesive tape as sticking tool. 6.根据权利要求1所述的一种固定家兔大动脉用以分析脂质斑块含量的方法,其特征在于:所述步骤E中透明板采用亚克力板。6 . The method for fixing the aorta of rabbits according to claim 1 to analyze the content of lipid plaques, characterized in that: in the step E, the transparent plate adopts an acrylic plate. 7 .
CN201911322530.7A 2019-12-20 2019-12-20 A method for immobilizing rabbit aorta for analyzing lipid plaque content Pending CN110967242A (en)

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Citations (6)

* Cited by examiner, † Cited by third party
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

Patent Citations (6)

* Cited by examiner, † Cited by third party
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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Application publication date: 20200407