The development of light sheet microscopy for automated sorting of zebrafish embryos

光片荧光显微镜是一种微创成像技术，可以用来观察活胚胎的发育过程。这个博士项目的目的是设计和制造一种与自动全胚胎分选仪兼容的光片荧光显微镜。其目标是提高斑马鱼胚胎成像的吞吐量和标准化，广泛应用于胚胎突变表型分析和药物筛选实验的分析。作为原理验证，将对已被设计为携带导致人类疾病的黏附G蛋白偶联受体基因adgrg6(也称为GPR126)的斑马鱼胚胎进行成像。成功的学生将使用转基因荧光标记来突出神经元和神经胶质细胞，以研究野生型和adgrg6突变胚胎中神经系统的发育和再生。他们还将能够测试通过药物筛选识别的拯救或拮抗小分子对发育中的胚胎和Adgrg6信号通路的影响。这个令人兴奋的跨学科项目结合了光学物理、发育生物学、药理学以及分子和遗传学方法。学生将在第一年设计和制造显微镜，获得激光、镜头和相机的使用培训和技能，以开发定制的成像平台。该显微镜将被安置在大学的沃尔夫森光学显微镜设施中，该设施为生物成像提供一整套高分辨率仪器。学生还将获得处理斑马鱼胚胎和使用高含量图像分析技术的经验。将有足够的机会为实验设计做出贡献，并在演示、教学和推广方面获得技能和经验。

Light-sheet fluorescence microscopy is a minimally-invasive imaging technique that can be used to observe living embryos as they develop. The aim of this PhD project is to design and build a light-sheet fluorescence microscope that is compatible with an automated whole-embryo sorting apparatus. The goal is to achieve improved throughput and standardisation of imaging of zebrafish embryos, with wide-ranging applications for the analysis of embryonic mutant phenotypes and for the analysis of drug screening experiments.As a proof-of-principle, imaging will be carried out on zebrafish embryos that have been engineered to carry human disease-causing genetic variants of an adhesion G protein-coupled receptor (aGPCR) gene, adgrg6 (also known as gpr126). The successful student will use transgenic fluorescent markers to highlight neurons and glia to study the development and regeneration of the nervous system in wild-type and adgrg6 mutant embryos. They will also be able to test the effects of rescuing or antagonist small molecules, identified through drug screens, on the developing embryo and the Adgrg6 signalling pathway.This exciting interdisciplinary project combines optical physics, developmental biology, pharmacology, and molecular and genetic approaches. The student will spend the first year designing and building the microscope, gaining training and skills in the use of lasers, lenses and cameras to develop a bespoke imaging platform. The microscope will be housed in the University's Wolfson Light Microscopy Facility, which provides a whole suite of high-resolution instruments for biological imaging. The student will also gain experience in working with zebrafish embryos, and in the use of high-content image analysis techniques. There will be ample opportunity to contribute to experimental design, and to gain skills and experience in presentation, teaching and outreach.