Method for preparing zebra fish thrombus model by utilizing ferric chloride

A technology of zebrafish and ferric chloride, applied in the field of medicine, can solve problems such as few methods and few studies on zebrafish thrombus model, and achieve the effects of cheap reagents, good film-forming effect and stable molding rate

Inactive Publication Date: 2021-04-30
叶繁全
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are few studies on zebrafish thrombus model, and there are not many alternative m...

Method used

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  • Method for preparing zebra fish thrombus model by utilizing ferric chloride
  • Method for preparing zebra fish thrombus model by utilizing ferric chloride
  • Method for preparing zebra fish thrombus model by utilizing ferric chloride

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Experimental program
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Embodiment 1

[0031] Embodiment 1: the preparation of animal model of the present invention

[0032] Experimental materials: DMSO and o-dianisidine were purchased from SIGMA; FeCl 3 , NaCl, KCl, CaCl 2 , MgSO 4 , sodium acetate, H 2 o 2 Purchased from Shanghai Jizhi Biochemical Technology Co., Ltd.; 6-well plate was purchased from Beijing Lvbaicao Technology Development Co., Ltd.

[0033] Experimental equipment: Leica DVM6 stereo microscope was purchased from Beijing Ruike Zhongyi Technology Co., Ltd.; constant temperature incubator JF-JGDW20-100 was purchased from Shanghai Jingfu Instrument Co., Ltd.

[0034] Breeding environment of zebrafish: The zebrafish used in the experiment is the AB line, and it is raised in Xiaoyu Protest Technology (Guangzhou) Co., Ltd. according to Westefield's breeding method. The water temperature is 28°C, the conductivity is 400-810μS / cm, and the pH value is 6.8-7.2. The light / dark cycle is 14h / 10h.

[0035] Experimental method: Select healthy and sexual...

Embodiment 2

[0037] Embodiment 2 This model is compared with the phenylhydrazine induction model

[0038] Experimental materials: DMSO and o-dianisidine were purchased from SIGMA; FeCl 3 , NaCl, KCl, CaCl 2 , MgSO 4 , sodium acetate, H 2 o 2 Purchased from Shanghai Jizhi Biochemical Technology Co., Ltd.; 6-well plate was purchased from Beijing Lvbaicao Technology Development Co., Ltd.; phenylhydrazine was purchased from Chengdu Dianchun Technology Co., Ltd., and phenylhydrazine was prepared with 100% ethanol to make a 1.5mmol / L mother solution.

[0039] Experimental equipment: Leica DVM6 stereo microscope was purchased from Beijing Ruike Zhongyi Technology Co., Ltd.; constant temperature incubator JF-JGDW20-100 was purchased from Shanghai Jingfu Instrument Co., Ltd.

[0040] Breeding environment of zebrafish: The zebrafish used in the experiment is the AB line, and it is raised in Xiaoyu Protest Technology (Guangzhou) Co., Ltd. according to Westefield's breeding method. The water tempe...

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Abstract

The invention relates to a method for preparing a zebra fish thrombus model by utilizing ferric chloride. The method comprises the following steps: 1) selecting about 30 2dpf zebra fishes which grow normally, transferring the zebra fishes into a 0.1%-1.1% ferric chloride solution prepared from embryo culture water, putting the zebra fishes into an incubator for culturing for 48 hours, sucking and discarding the culture water, and carrying out staining away from light for 15 minutes in a 6-micropore plate by using an o-dianisidine staining solution; 2) removing the staining solution, and performing quick cleaning with DMSO (dimethylsulfoxide) for three times; (3) transferring the zebra fishes into a new 6-micropore plate, and adding DMSO with the same volume; 4) placing the zebra fishes on a glass slide, and photographing the zebra fishes under a microscope and storing the zebra fishes; and 5) calculating the zebra fish trunks and thrombus staining area by using Image-ProPlus6, and utilizing (thrombus staining area/trunk area)*100% as an index of thrombosis occurring degree. The method has the beneficial effects that the reagent is cheap, the model forming rate is stable, and the model forming effect is good.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to a method for preparing a zebrafish thrombus model by using ferric chloride. Background technique [0002] Thrombosis is caused by multiple genetic and environmental factors, and can occur in arteries and veins. Venous thrombus is mostly red thrombus, which can cause limb swelling and pain. Venous thrombus can enter the pulmonary circulation and cause pulmonary embolism. In severe cases, there will be dyspnea. Even death. Arterial thrombus is mostly white thrombus, which will cause ischemic changes in limbs and important organs. Arterial thrombus can block distal arteries along the blood flow, causing necrosis of limbs or organs. Thrombosis is more harmful to the human body. Active treatment, at present, due to social pressure, serious environmental pollution, unhealthy lifestyle and other reasons, the trend of rejuvenation of thrombosis is increasing day by day. [0003] Zebr...

Claims

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Application Information

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IPC IPC(8): A01K61/10A61K49/00
CPCA01K2227/40A01K2267/03A61K49/0008A01K61/10Y02A40/81
Inventor 叶繁全
Owner 叶繁全
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