NanoDrill - A new versatile research tool - high spatial resolution light activated molecular nanomachines

未来生物医学和基础细胞生物学的一个重要需求是有针对性地在体内破坏选定的细胞和细胞类型。癌症影响到每个国家，无论其财政状况、医疗基础设施或高技能专家的可用性如何，癌症每年在全球造成820万人死亡。我们对未来的愿景是开发一系列光激活的分子纳米机器——纳米钻头——选择性地靶向癌细胞并安全地根除它们。目前，这只能通过高度侵入性的外科手术或放射治疗来实现，而这些手术或放射治疗往往是有害的。我们已经证明，我们的纳米钻（分子纳米机器，MNMs）可以使用少量的紫外线激活光选择性地破坏细胞。我们研究的下一个阶段是四倍的。首先，我们必须在现有的显微镜系统上启用多双光子（2PE）激活，使用生物安全的近红外（NIR）光进行活细胞研究。其次，扩展细胞类型和病理状况的特定纳米钻的家族，以允许更广泛的癌细胞研究。第三，设计可以使用普通可见光激活的纳米钻，以符合生物医学研究中已经建立的协议。第四，创造一种新型的纳米钻，这种纳米钻将首先被靶向癌细胞吸收，然后允许从内部触发根除。这将促进更可控的单细胞精密研究工具。初步研究表明，我们的MNMs可以用2PE成功激活。为了继续这项研究，我们需要一种快速、高分辨率的活细胞多光子显微镜（MPM）来对活细胞进行成像。不幸的是，由于其性质和独特的仪器要求，现成的MPM系统的成本在50万英镑左右。然而，我们建议开发一个合适的定制系统，通过调整现有设备的一小部分成本。我们将装备我们的徕卡SP5 II共聚焦系统与可调的高重复率的最先进的近红外激光器。这种实验修改将使我们能够直接比较活细胞分子纳米机器在UV和2PE激发下的激活，并且超出了标准的现成显微镜的能力。我们已经购买了附加近红外激光器所需的所有相关辅助光学元件。我们将仪器构建推进到一个阶段，在采购所需的激光系统后，一个简单的“即插即用”方法将在几周内在我们的仪器上产生多光子升级。一旦完全开发，它将形成一种新的高精度生物研究工具，并为伤口愈合和肿瘤/癌症进展研究开辟新的视野。它可能为对化疗药物耐药的癌症（如转移性乳腺癌）的非侵入性治疗铺平道路。这个项目有可能在全球范围内改变游戏规则。

An important need for future bio-medical and fundamental cell biology is the targeted in vivo destruction of selected cells and cell types. Cancer affects every country regardless of financial status, medical infrastructure or the availability of highly-skilled specialists and is responsible for 8.2 million deaths/year worldwide. Our vision for the future is to develop a series of light-activated molecular nanomachines - Nano-drills - to target cancerous cells selectively and safely eradicate them. Currently this can only be facilitated using highly invasive surgical or radiotherapeutic procedures that are often harmful to administer.We have already demonstrated that our Nano-drills (molecular Nano-Machines, MNMs) can selectively destroy cells using small quantities of ultraviolet activation light. The next phase of our research is four-fold. First, we must enable multi-two-photon (2PE) activation on our existing microscope system using biologically safe near-infra-red (NIR) light for live-cell studies. Second, extend the family of cell type and pathological condition specific nano-drills to allow a wide range of cancer cells to study. Third, design nano-drills that can be activated using plain visible light in order to comply with already established protocols in biomedical research. Fourth, create a new breed of nano-drills that will first be internalised by the targeted cancer cells and subsequently allowing eradication to be triggered them from within. This will promote a more controlled single-cell precision research tool.Preliminary studies showed that our MNMs can be successfully activated using 2PE. In order to continue this research, we need a fast, high resolution live cell capable Multi-Photon Microscope (MPM) to image live cells. Unfortunately, due to the nature and unique instrumental requirement an off the shelf MPM system costs in the region of £500,000. However, we propose to develop a suitable custom system by adapting existing equipment at fraction of the cost. We will equip our Leica SP5 II confocal system with a tuneable high repetition rate state-of-the-art NIR laser. This experimental modification will allow us to directly compare live-cell molecular nanomachine activation with both UV and 2PE excitation and exceed the capabilities of a standard off-the-shelf microscope. We have already purchased all associated auxiliary optical components needed for attaching the NIR laser. We advanced the instrument build to a stage, where after sourcing the required laser system a simple 'plug and play' approach will yield the multi-photon upgrade on our instrument within a few weeks. Once fully developed it could form a new extremely high precision biological research tool and opens new horizons toward wound healing and tumour/cancer progression studies. It could pave the way towards a non-invasive treatment of chemotherapeutic agent resistant cancers, such as metastatic breast cancer. This project has a potential game-changing impact on a global scale.