MEGA-FLIM: quantum technologies for megapixel time-resolved imaging and control across biological scales

胚胎、器官和肿瘤是由许多细胞组成的，这些细胞相互作用，并跨越几个细胞的距离进行交流。虽然在显微镜下研究单个细胞是例行公事，但由于它们的大小、光散射特性和复杂性，研究集体细胞要困难得多。荧光寿命成像(FLIM)和FRET(Forster共振能量转移)使用能量转移的原理，当光粒子(光子)从受激发的荧光供体分子跳到附近的受体，从而改变供体的荧光寿命时发生的能量转移。Flim被用来通过测量这种寿命变化来测量活细胞内部密切的分子相互作用。我们将把物理、工程、计算和生物结合起来，构建一个具有极高时空分辨率的新的光学微观系统。我们将我们的系统称为超级薄膜，我们将使用它来研究更大的细胞集体，以发现细胞如何对机械和化学信号做出反应进行交流和组织。Mega-Flim将允许在比以前可能的更大的区域内更快地收集光信号。我们还将开发一种技术，利用光来控制这些集体的细胞行为，这项技术被称为光遗传学。我们由光学物理学家、生物工程师和生物学家组成的独特团队是打破当前障碍的理想人选，导致在每个领域都有里程碑式的发现。为什么我们需要一种新的薄膜显微镜系统？商业系统缺乏同时允许：-快速采集(0.1秒或更快)，以便能够以高分辨率(100万像素或更高)在大范围内对活细胞或胚胎进行实时测量，从而以高时间分辨率(50-100皮秒)对整个细胞环境和大集体进行成像，从而能够精确地区分生命周期-双光子激发，以便能够精确地全3D重建细胞集体。-宽视场光遗传激活(光控制的细胞行为)，以便在存在跨多个部位的复杂激活刺激的情况下研究集体的动力学。这个新系统将解决什么问题？它将产生什么影响？-Mega Flim将提供一个系统，允许我们以分子分辨率询问生命系统，并发现细胞在迁移时如何使用化学和机械信号来控制集体信号。这种引导使细胞能够识别彼此和其他细胞类型，并在3个维度上形成复杂的模式(如器官或胚胎)。-我们的新系统将具有巨大的商业价值，因为它将提高对细胞行为的成像和光遗传控制的能力。-通过建立一个系统，我们可以发现控制细胞集体行为的新途径，我们将获得可靠和可预测地控制集体细胞行为的能力。这门学科被称为合成生物学，非常适合用于医疗和商业用途，用于构建器官/芯片上肿瘤系统，或创建用于药物发现的生理相关系统。

Embryos, organs and tumours are composed of many cells, which interact with each other and communicate across distances of several cells. While it is routine to study single cells under a microscope, it is much more difficult to study collectives due to their size, light scattering properties and complexity. Fluorescence lifetime imaging (FLIM) and FRET (Forster resonance energy transfer) use the principles of energy transfer that occur when a light particle (photon) jumps from an excited fluorescent donor molecule to a nearby acceptor, thus changing the fluorescence lifetime of the donor. FLIM is used to measure close molecular interactions inside of living cells by measuring this lifetime change. We will combine physics, engineering, computation and biology to build a new light microscopic system, with extremely high spatial and temporal resolution. We call our system MEGA-FLIM and we will use it to study larger cell collectives of cells to discover how cells communicate and organise in response to both mechanical and chemical signals. MEGA-FLIM will allow much faster collection of light signals across a much larger field than previously possible. We will also develop technology to use light to control cell behaviour across these collectives using the technique called optogenetics. Our unique team of optical physicists, bioengineers and biologists is ideally placed to break down current barriers, leading to landmark discovery in each of these fields.Why do we need a new FLIM microscope system?Commercial systems are lacking that allow, simultaneously: - fast acquisition (0.1 second or faster) so as to allow real-time measurements in live cells or embryos- across a widefield area with high resolution (1 million pixels or higher), so as to allow imaging of the full cell environment and large collectives- with high time resolution (50-100 pico seconds), so as to allow precise discrimination of lifetimes- two-photon excitation, so as to allow precise full 3D reconstruction of cell collectives.- widefield optogenetic activation (light-controlled cell behaviour), so as to allow study of the dynamics of collectives in the presence of complex activation stimuli that act across multiple sites.What problems will this new system solve and what impact will it have?-MEGA FLIM will provide a system that will allow us to interrogate living systems at molecular resolution and discover how cells collectively signal using both chemical and mechanical signals to steer when they migrate. This kind of steering allows cells to recognise each other and other cell types and to form complex patterns in 3 dimensions (like in an organ or an embryo).-Our new system will be of great commercial interest, as it will advance capabilities in imaging and optogenetic control of cell behaviour with light.-By building a system whereby we can discover new pathways governing how cells behave in collectives, we will gain the ability to reliably and predictably control collective cell behaviour. This discipline, known as synthetic biology, is highly desirable for medical and commercial use in building organ/tumour-on-chip systems or creating physiologically relevant systems to use in drug discovery.