Label-free, Real-time, Spatial-resolution (LRS) immunoassay: 2D mapping of extracellular signalling molecules

人类和动物组织中的单个细胞通过特定的化学和生物分子信号相互交流，这些信号从细胞释放到当地环境中。这些信号在控制细胞活动方面起着关键作用，包括调节细胞的生长、功能、运动和死亡。这些信号共同调节组织的形成、功能和修复。发明一种可以在健康和疾病组织中创建这些分子信号的高清晰度地图的技术，将为了解组织如何工作以及它们在发育、疾病和衰老过程中的变化提供宝贵的见解。包括酶免疫分析(ELISA)在内的传统分析技术已经在医院诊断、家庭怀孕检测试剂盒和实验室实验中使用了40多年。虽然这项技术和其他相关技术已经能够识别和量化免疫反应、肿瘤生长、发育和组织功能中的关键信号分子，但它们没有提供关于这些分子在组织中的哪里分泌、分子如何在组织中扩散或何时扩散的信息。因此，他们无法解决这些信号分子实际如何工作的关键机制问题，细胞何时对这些信号做出反应以及为什么。在这项提议中，我们的目标是开发、测试和使用一种新的分析技术，该技术将提供多细胞组织中的分子信号的实时映射，使研究人员和临床医生能够回答这些关键问题。该工具基于硅光子晶体纳米结构，类似于最初为高速电信应用开发的纳米结构。我们最近已经证明，光子晶体对当地环境的敏感性可以被用来检测非常低浓度的蛋白质。检测是实时进行的，而光子晶体的小尺寸使分子检测具有很高的空间分辨率。在这项提案中，我们将通过解决生物学中一个关键的未知问题来展示这项新技术的潜力，该问题涉及称为白细胞介素素的信号蛋白在免疫反应中的作用。

Individual cells within human and animal tissues communicate with each other through specific chemical and bio-molecular signals that are released from cells into their local environment. These signals have critical roles in controlling cellular activity including regulating cell growth, function, movement and death. Together these signals regulate how tissues form, function and are repaired. Inventing a technology that can create high definition maps of these molecular signals in both healthy and diseased tissues would provide invaluable insights into how tissues work and how they change during development, disease and during the aging process.Conventional analytical techniques including enzymatic immune assays (ELISA) have been used in hospital diagnostics, home pregnancy detection kits and laboratory experiments for over 40 years. While this and other related techniques have been able to identify and quantify key signalling molecules in immune responses, tumour growth, development and tissue function they provide no information regarding where in the tissue these molecules are secreted, how the molecules diffuse in the tissue or indeed when. As a result, they are unable to address key mechanistic questions about how these signalling molecules actually work, when cells respond to these signals and why.In this proposal we aim to develop, test and use a new analytical technology that will provide real-time mapping of molecular signals in multicellular tissues, enabling researchers and clinicians to answer these critical questions. The tool is based on a silicon photonic crystal nanostructure similar to those developed originally for applications in high speed telecommunications. We have shown recently that the sensitivity of the photonic crystals to the local environment can be exploited to detect very low concentrations of proteins. The detection occurs in real-time and the small size of the photonic crystal enables molecular detection with very high spatial resolution. In this proposal, we will demonstrate the potential of this new technology by addressing a key unknown question within biology regarding the role of signalling proteins called interleukins in the immune response.