Developing next generation bioimaging/biophotonics tools to dissect the immunological synapse in single cells, one molecule at a time

该项目旨在开发/应用新的生物成像/生物光子学工具，以深入了解跨多个长度/时间尺度的复杂生物过程，从单分子到亚细胞结构，细胞和细胞群体。学生需要渴望拥抱从生物物理学到细胞/分子生物学的跨学科科学，并将向这些领域的世界级团队学习。该学生将在约克大学生物系Mark Leake教授（http://www.single-molecule-biophysics.org/）的团队中工作，并由Nathalie Signoret博士（https://www.york.ac.uk/cii/staff/academic/signoret/）共同监督。本项目由国家物理实验室（NPL）、英国国家测量研究所支持，由NPL生物计量学组Mike Shaw （http://www.npl.co.uk/people/mike-shaw）联合监督；该学生将在位于伦敦西南部特丁顿的国家物理实验室主站工作12个月。该项目的重点是了解免疫突触（IS）如何形成和调节。IS定义了免疫细胞交流的接触过程，在T细胞-巨噬细胞相互作用的情况下，这对于宿主对感染的反应至关重要。然而，关于IS形成/调节的关键机制细节尚不清楚，因为关注的是过于狭窄的长度/时间尺度制度。您将开发跨长度尺度成像/光子学工具，以深入了解细胞中IS形成/调节的动态复杂性，涉及体内快速单分子检测和定量，并结合纳米尺度空间精度（见Miller等人2018 PMID 29872430； Wollman等人2017 PMID:28841133； Kasprowicz等人PMID:29789661； Shaw等人PMID 25839410）。该项目旨在增加对趋化因子刺激如何用于调节is形成的新理解，重点是趋化因子受体CCR5，在转移到更具挑战性的原代巨噬细胞和T细胞形成is之前，对模型细胞系进行初步研究，以解决基础CCR5配体行为，探索趋化因子受体-配体相互作用的动力学，趋化因子与细胞表面蛋白聚糖的结合，以及is形成过程中局部膜脂和蛋白质的相互作用。

This project aims to develop/apply new bioimaging/biophotonics tools to gain insight into complex biological processes across multiple length/timescales, from single molecules through to subcellular structures, cells and cellular populations. The student will need to be eager to embrace interdisciplinary science from biophysics through to cell/molecular biology, and will learn from world-class teams in these areas. The student will work in the Department of Biology, University of York, in the team of Prof Mark Leake (http://www.single-molecule-biophysics.org/), co-supervised by Dr Nathalie Signoret (https://www.york.ac.uk/cii/staff/academic/signoret/). This project is supported by the National Physical Laboratory (NPL), the UK National Measurement Institute, jointly supervised by Mike Shaw (http://www.npl.co.uk/people/mike-shaw) from NPL's Biometrology group; the student will spend 12 months at NPL's main site in Teddington, SW London. The project focuses on understanding how the Immunological Synapse (IS) forms and is regulated. The IS defines contact processes of immune cells communication, which in the case of T cell-macrophage interactions is crucial for a host response to infection. However, key mechanistic details concerning IS formation/regulation are unknown due to focusing on too narrowly defined length/timescale regimes. You will develop cross lengthscale imaging/photonics tools to enable insight into the dynamic complexities of IS formation/regulation in cells, involving rapid single-molecule detection and quantification in vivo combined with nanoscale spatial precision (see Miller et al 2018 PMID 29872430; Wollman et al 2017 PMID:28841133; Kasprowicz et al PMID:29789661; Shaw et al PMID 25839410).The project aims to add new understanding in how chemokine stimulation is used for regulation of IS formation, focusing on chemokine receptor CCR5, with initial investigations on model cell-lines to address basal CCR5-ligand behaviour before moving to more challenging primary macrophages and IS formation with T cells, probing dynamics of chemokine receptor-ligand interactions, chemokine binding to cell-surface proteoglycan, and local membrane lipids and proteins interactions during IS formation.