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首页> 外文期刊>Transgenic research >Fluorescence imaging preparation methods for tissue scaffolds implanted into a green fluorescent protein porcine model
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Fluorescence imaging preparation methods for tissue scaffolds implanted into a green fluorescent protein porcine model

机译：用于绿色荧光蛋白猪模型的组织支架的荧光成像制备方法

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Green fluorescent protein (GFP) animal models have become increasingly popular due to their potential to enhance in vivo imaging and their application to many fields of study. We have developed a technique to observe host tissue integration into scaffolds using GFP expressing swine and fluorescence imaging. Current fluorescence imaging preparation methods cannot be translated to a full GFP animal model due to several challenges and limitations that are investigated here. We have implanted tissue scaffolds into GFP expressing swine and have prepared explanted scaffolds for fluorescence imaging using four different methods including formalin fixation and paraffin embedding, vapor fixation, freshly prepared paraformaldehyde fixation, and fresh frozen tissue. Explanted scaffolds and tissue were imaged using confocal microscopy with spectral separation to evaluate the GFP animal model for visualization of host tissue integration into explanted scaffolds. All methods except fresh frozen tissue induced autofluorescence of the scaffold, preventing visualization of detail between host tissue and scaffold fibers. Fresh frozen tissue preparation allowed for the most reliable visualization of fluorescent host tissue integration into non-fluorescent scaffolds. It was concluded that fresh frozen tissue preparation is the best method for fluorescence imaging preparation when using scaffolds implanted into GFP whole animal models.
机译：绿色荧光蛋白（GFP）动物模型因其增强体内成像的潜力及其在许多研究领域中的应用而变得越来越受欢迎。我们已经开发了一种使用表达GFP的猪和荧光成像技术观察宿主组织整合入支架的技术。由于此处研究的一些挑战和局限性，当前的荧光成像制备方法无法转化为完整的GFP动物模型。我们已经将组织支架植入表达GFP的猪中，并使用四种不同的方法（包括福尔马林固定和石蜡包埋，蒸气固定，新鲜制备的多聚甲醛固定以及新鲜的冷冻组织）准备了用于荧光成像的外植支架。使用共聚焦显微镜和光谱分离对移植的支架和组织成像，以评估GFP动物模型，以可视化宿主组织整合到移植的支架中。除新鲜的冷冻组织外，所有方法均会诱导支架自发荧光，从而阻止宿主组织和支架纤维之间细节的可视化。新鲜的冷冻组织制备物可以最可靠地显示荧光宿主组织整合到非荧光支架中的情况。结论是，新鲜的冷冻组织制备物是使用植入GFP完整动物模型的支架进行荧光成像制备的最佳方法。

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