Development of the worlds first Live Cell Nanoscope

自世纪末发明第一台复合光学显微镜以来，光学显微镜已经取得了几项关键性的进展。从那时起，我们已经经历了几次迭代，从增强放大，到提高分辨率（衍射极限），靶向荧光，高信噪比共聚焦和最近的超分辨率成像。就在2014年，诺贝尔化学奖被授予“超分辨荧光显微镜的发展”。2015年，我们推出了一款革命性的超分辨荧光显微镜，它不仅打破了传统的分辨率限制（阿贝极限），而且还允许非常快速和温和的成像，因此使其成为超分辨“活细胞成像”的最先进工具。然而，这种显微镜仍然局限于通常所说的光学显微镜的SIM/ISM分辨率极限，近似为分辨率=（0.61\*？）/（2 * 不适用）。对于排放？525 nm和数值孔径（NA）为1.49的物镜，最佳可实现的分辨率为107 nm。虽然这确实为细胞生物学中发生的一些最小的相互作用提供了一个窗口，但从纳米尺度（= 50 nm）下的解析中可以获得的信息量的增加是指数级的。（SMLM、STED、CLEM）所有这些都需要固定生物样本，因此不允许您看到在关键生物事件（如细胞）期间发生的大量亚细胞相互作用。分裂，蛋白质与蛋白质的相互作用，细胞与病毒和细菌的相互作用，细胞收缩等。作为该项目的一部分，我们建议开发世界上第一个基于荧光的纳米显微镜，使活细胞成像。还应该注意的是，开发的仪器不仅适用于科学研究领域;先进的光学显微镜不再局限于科学实验室，正用于各种临床和工业应用。这种高速的活细胞成像纳米镜将为医学筛选中的筛选应用开辟新的可能性，例如用于诊断血小板颗粒紊乱，用于药物开发的高含量筛选（HCS），例如mRNA疫苗的开发和用于高密度原位测序的基因测序，以空间解析RNA/DNA/蛋白质，用于开发个性化药物。

Light microscopy has made several key advancements since the invention of the first compound light microscope in the late 16th century. Since then we have moved through several iterations from enhanced magnification, to improved resolution (diffraction limited), targeted fluorescence, high signal to noise confocal and most recently super resolution imaging.As recently as 2014 the Nobel Prize in Chemistry was awarded for 'the development of super-resolved fluorescence microscopy'. In 2015 we launched a revolutionary super resolved fluorescent microscope which not only broke the traditional limits of resolution (Abbe's limit) but also allowed for very fast and gentle imaging, therefore making it the most advanced tool for super resolved 'Live Cell Imaging.'However, this microscope was still limited to what is commonly referred to as the SIM/ISM limit of resolution for a light microscope, which is approximated as Resolution = (0.61\*?)/(2\*NA). So for an emission ? of 525nm and an objective with Numerical Aperture (NA) of 1.49, the best achievable resolution would be 107nm. Whilst this does offer a window into some of the smallest interactions which take place within cell biology, the increase in the amount of information that can be gained from resolving down at the nanoscopy scale (=50nm) is exponential.Currently the only tools available for fluorescence based nanoscopy (SMLM, STED, CLEM) all require the biological specimen to be fixed and therefore do not allow you to see the huge amount of sub cellular interactions which occur during critical biological events such as cell division, protein to protein interaction, cell interactions with viruses and bacteria, cell contraction, etc...What we are proposing to develop as part of this project is the worlds first fluorescent based nanoscope which enables live cell imaging.It should also be noted that the developed instrument would not only be suitable for the scientific research field; advanced light microscopes are no longer confined to scientific laboratories and are being used for a variety of clinical and industrial (pharmaceutical) applications, from medical screening to drug development.Such a high speed, live cell imaging nanoscope would open up new possibilities for screening applications in medical screening such as for the diagnosis of platelet granule disorder, high content screening (HCS) for drug development such as in the development of mRNA vaccines and gene sequencing for high density in-situ sequencing to spatially resolve RNA/DNA/Proteins for development of personalised medicines.