The invention relates to a zebrafish wound detection method, which comprises the following steps: (1) preparation of zebrafish: cutting zebrafish and making wound site; (2) preparation of test solution: preparation of fluorescent probe reserve solution and control probe reserve solution, and dilution of two reserve solutions into fluorescent probe test solution and control probe test solution by PBS buffer solution respectively; (3) preparation of test solution for zebrafish; Wound detection: Two kinds of test solutions were used as water for zebrafish culture, and continuous culture. Fluorescence imaging of fluorescence probe and control probe in zebrafish was collected and recorded by fluorescence microscope. The two groups of fluorescence imaging were compared to judge the situation of zebrafish wound. Compared with the prior art, the invention can not only detect the wound condition, but also can be used to monitor the wound healing condition, and the detection probe used has the advantages of low cytotoxicity, simple method, strong practicability, fast and accurate, etc.
【技术实现步骤摘要】
一种斑马鱼伤口检测方法
本专利技术属于生物检测
,具体涉及一种斑马鱼伤口检测方法。
技术介绍
斑马鱼属于脊椎动物,和人类基因极为相似,具有高达87%的同源性,可作为一种理想的动物模型(MEEKER,NathanD.;TREDE,NikolausS.Immunologyandzebrafish:spawningnewmodelsofhumandisease.Developmental&ComparativeImmunology,2008,32.7:745-757)。此外,斑马鱼体型小,易于养殖,产卵量高且繁殖周期短,这意味着大大降低了使用成本。斑马鱼的受精方式为体外受精,生长发育全程透明,有利于实时观察病原体和损伤。因而,斑马鱼可作为理想的炎症药物筛选模型。如通过构建由绿色荧光蛋白标记巨噬细胞的转基因斑马鱼系,筛选出其中绿色荧光蛋白表达稳定的斑马鱼,进一步切割尾部构建巨噬细胞聚集的炎症模型,通过追踪炎症细胞迁移,监测伤口愈合情况(秦帅,陈希,and孔德明."构建由绿色荧光标记巨噬细胞的转基因斑马鱼模型."贵阳中医学院学报33.5(2011):133-135.)。类似的,通过构建中性粒细胞转基因斑马鱼系,可建立炎症反应的体内遗传分析模型(RENSHAW,StephenA.,etal.Atransgeniczebrafishmodelofneutrophilicinflammation.Blood,2006,108.13:3976-3978),从而追踪炎症反应。这一类构建炎症模型的方法基于机体受到刺激发生炎症反应,中性粒细胞和巨噬细胞向炎症部位聚集 ...
【技术保护点】
1.一种斑马鱼伤口检测方法,其特征在于,该方法包括以下步骤:(1)斑马鱼的准备:将斑马鱼割伤,制造创伤部位;(2)测试溶液的配制:制备荧光探针储备液,并利用PBS缓冲液将储备液稀释成荧光探针测试溶液;(3)斑马鱼伤口的检测:将测试溶液作为斑马鱼的培养用水,持续培养,并利用荧光显微镜采集记录荧光探针在斑马鱼体内的荧光成像,判断斑马鱼伤口情况。
【技术特征摘要】
1.一种斑马鱼伤口检测方法,其特征在于,该方法包括以下步骤:(1)斑马鱼的准备:将斑马鱼割伤,制造创伤部位;(2)测试溶液的配制:制备荧光探针储备液,并利用PBS缓冲液将储备液稀释成荧光探针测试溶液;(3)斑马鱼伤口的检测:将测试溶液作为斑马鱼的培养用水,持续培养,并利用荧光显微镜采集记录荧光探针在斑马鱼体内的荧光成像,判断斑马鱼伤口情况。2.根据权利要求1所述的一种斑马鱼伤口检测方法,其特征在于,配制对照探针储备液,并利用PBS缓冲液将储备液稀释成对照探针测试溶液,作为斑马鱼培养用水,持续培养,将两组荧光成像进行对比。3.根据权利要求2所述的一种斑马鱼伤口检测方法,其特征在于,所述的对照探针为对叔丁基苯酚修饰的七甲川花菁染料,结构式为:4.根据权利要求1所述的一种斑马鱼伤口检测方法,其特征在于,所述步骤(2)中的荧光探针为4-碘代-2-丁醇修饰的七甲川花菁染料,结构式为:5.根据权利要求1所述的一种斑马鱼伤口检测方法,其特征在于,所述步骤(2)中的探针储备液通过将探针固体用二甲基亚砜溶剂溶解、超声制备得到,探针测试溶液里溶剂二甲基亚砜的含量不高于...
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