Department of Molecular Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA.
Nat Protoc. 2011 Feb;6(2):229-41. doi: 10.1038/nprot.2010.170. Epub 2011 Feb 3.
Zebrafish are a useful vertebrate model for the study of development, behavior, disease and cancer. A major advantage of zebrafish is that large numbers of animals can be economically used for experimentation; however, high-throughput methods for imaging live adult zebrafish had not been developed. Here, we describe protocols for building a light-emitting diode (LED) fluorescence macroscope and for using it to simultaneously image up to 30 adult animals that transgenically express a fluorescent protein, are transplanted with fluorescently labeled tumor cells or are tagged with fluorescent elastomers. These protocols show that the LED fluorescence macroscope is capable of distinguishing five fluorescent proteins and can image unanesthetized swimming adult zebrafish in multiple fluorescent channels simultaneously. The macroscope can be built and used for imaging within 1 day, whereas creating fluorescently labeled adult zebrafish requires 1 hour to several months, depending on the method chosen. The LED fluorescence macroscope provides a low-cost, high-throughput method to rapidly screen adult fluorescent zebrafish and it will be useful for imaging transgenic animals, screening for tumor engraftment, and tagging individual fish for long-term analysis.
斑马鱼是一种用于研究发育、行为、疾病和癌症的有用的脊椎动物模型。斑马鱼的一个主要优势是可以经济地使用大量动物进行实验；然而，用于对活体成年斑马鱼进行高通量成像的方法尚未开发。在这里，我们描述了构建发光二极管（LED）荧光体视显微镜的方案，并描述了如何使用它同时对 30 只以上表达荧光蛋白的转基因成年动物、荧光标记的肿瘤细胞移植的成年动物或标记有荧光弹性体的成年动物进行成像。这些方案表明，LED 荧光体视显微镜能够区分五种荧光蛋白，并能够同时对未麻醉的游动成年斑马鱼进行多个荧光通道成像。体视显微镜可以在 1 天内构建和使用进行成像，而创建荧光标记的成年斑马鱼则需要 1 小时到几个月的时间，具体取决于所选择的方法。LED 荧光体视显微镜提供了一种低成本、高通量的方法来快速筛选荧光斑马鱼成年鱼，它将对成像转基因动物、筛选肿瘤移植以及对单个鱼类进行长期分析非常有用。
