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在体内有针对性地销毁选定的细胞和细胞类型。癌症影响每个国家，无论其经济状况、医疗基础设施或高技能专家的可获得性如何，全球每年有820万人死于癌症。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，这只能通过高侵入性的手术或放射治疗程序来促进，这些程序往往对管理有害。我们已经证明，我们的纳米钻头(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。我们下一阶段的研究有四个方面。首先，我们必须在我们现有的显微镜系统上使用生物安全的近红外(NIR)光来进行活细胞研究，从而实现多双光子(2PE)激活。第二，扩大针对细胞类型和病理条件的特定纳米钻探，以允许广泛的癌细胞研究。第三，设计可以用普通可见光激活的纳米钻头，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头，它将首先被目标癌细胞内化，然后允许从内部触发根除它们。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MNM可以成功地使用2PE激活。为了继续这项研究，我们需要一台快速、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，由于性质和独特的仪器要求，现成的MPM系统成本约为500,000 GB。然而，我们建议通过调整现有设备来开发合适的定制系统，成本仅为其一小部分。我们将为徕卡SP5 II共焦系统配备一台可调谐的最先进的高重复频率近红外激光器。这种实验性的改进将使我们能够直接比较活细胞分子纳米机器的激活与紫外线和2PE激发，并超过标准的现成显微镜的能力。我们已经购买了连接近红外激光器所需的所有相关辅助光学部件。我们将仪器的构建推进到了一个阶段，在采购所需的激光系统后，简单的即插即用方法将在几周内为我们的仪器带来多光子升级。一旦完全开发，它可能形成一种新的极高精度的生物学研究工具，并为伤口愈合和肿瘤/癌症进展研究开辟新的视野。它可能为化疗耐药癌症的非侵入性治疗铺平道路，例如转移性乳腺癌。该项目在全球范围内具有潜在的改变游戏规则的影响。

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.