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A Super-resolution multiphoton and dynamic STORM imaging facility

超分辨率多光子和动态风暴成像设施

基本信息

批准号：
BB/M012034/1
负责人：
Nicholas Hartell
金额：
$ 81.3万
依托单位：
University of Leicester
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2015
资助国家：
英国
起止时间：
2015 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FM012034%2F1
关键词：
Super resolution multiphoton dynamic STORM

项目摘要

Recent advances in fluorescence microscopy have led to significant improvements in our ability to locate and discriminate small objects. Super-resolution is the term used for technologies that can resolve objects that are smaller or closer together than was previously thought possible according to the laws of Physics. It is now possible to see objects with nanometre resolution and precision as well as image at depth in living tissue and examine rapid biological events at high speed. Achieving all of these improvements at the same time is the "holy grail" of optical microscopy but this has proved practically difficult. At the University of Leicester, we have developed a new hybrid technology called SuperRAMP that allows high speed, multicolour, multiphoton, super-resolution imaging deep in living tissues.Super-resolution, Random Access MultiPhoton microscopy (SuperRAMP) is a unique technique that combines patterned illumination with mathematical methods to pinpoint discrete objects with nanometre precision at very high speeds. Devices that use sound waves passing across crystals are used to project patterns of infra-red light, which can pass through specimens better than visible wavelengths of light and produce better penetration into deep tissues. These patterns are processed mathematically to produce high-resolution images. SuperRAMP improves both the lateral (xy) and depth (z) resolution by 2-3-fold compared to standard microscopes to an absolute lateral resolution of 120 nm. It can collect two colour images at speeds of around 10 frames per second and from super-resolved points of interest at thousands of samples per second. No other microscope available today can operate at this resolution, at depth, and at this speed in living tissues. SuperRAMP will be combined with a second, complementary method called dSTORM to provide even higher resolution. dSTORM uses the same mathematical techniques used for SuperRAMP microscopy but without scanning. Although it takes longer to create an image, it has a higher resolution of 20 nm.We will develop a super-resolution, multi-user facility that will make these exciting technologies available to researchers at Leicester and to the wider community. This will help us to drive academic excellence amongst existing BBSRC funded researchers in projects that range from studies of the synthesis of new proteins, the processes of cell division, mechanisms of cell-cell communication in the central nervous system, the neurological bases for development and behaviour - including circadian clocks that control daily rhythms - in model animal systems including locusts, zebra fish, fruit flies, rats and mice.This facility will support research collaborations within the Midlands (Nottingham, Leicester and Warwick) and further afield (Cambridge) and allow us to strengthen our commercial impact by developing a show case facility for functional, super-resolution imaging. Having developed this revolutionary technology, we are uniquely placed to establish a world-leading centre of excellence for functional, super-resolution imaging.

荧光显微镜的最新进展大大提高了我们定位和辨别小物体的能力。“超分辨率”指的是一种技术，它可以分辨出比以前根据物理定律所能想象的更小或更近的物体。现在可以以纳米分辨率和精度看到物体，也可以在活体组织中进行深度成像，并可以高速检查快速的生物事件。同时实现所有这些改进是光学显微镜的“圣杯”，但事实证明这在实践中是困难的。在莱斯特大学，我们开发了一种叫做SuperRAMP的新混合技术，它可以在活体组织深处进行高速、多色、多光子、超分辨率的成像。超分辨率，随机存取多光子显微镜（SuperRAMP）是一种独特的技术，结合了图案照明和数学方法，以非常高的速度以纳米精度精确定位离散物体。利用声波穿过晶体的装置被用来投射红外光的图案，红外光比可见光波长的光更能穿过标本，并能更好地穿透深层组织。这些模式经过数学处理后产生高分辨率的图像。与标准显微镜相比，SuperRAMP将横向（xy）和深度(z)分辨率提高了2-3倍，绝对横向分辨率为120 nm。它可以以大约每秒10帧的速度收集两张彩色图像，并以每秒数千个样本的速度从超分辨率兴趣点收集图像。目前市面上还没有其他显微镜能够以这样的分辨率、深度和速度对活体组织进行观察。SuperRAMP将与另一种称为dSTORM的互补方法相结合，以提供更高的分辨率。dSTORM使用与SuperRAMP显微镜相同的数学技术，但没有扫描。虽然创建图像需要更长的时间，但它的分辨率更高，达到20纳米。我们将开发一个超分辨率、多用户的设备，将这些令人兴奋的技术提供给莱斯特大学的研究人员和更广泛的社区。这将帮助我们在现有的BBSRC资助的研究人员中推动学术卓越，这些项目包括研究新蛋白质的合成，细胞分裂过程，中枢神经系统中细胞-细胞通信机制，发育和行为的神经学基础-包括控制日常节律的生物钟-在模型动物系统中，包括蝗虫，斑马鱼，果蝇，大鼠和小鼠。该设施将支持中部地区（诺丁汉、莱斯特和沃里克）和更远地区（剑桥）的研究合作，并通过开发功能超分辨率成像的展示设施，使我们能够加强我们的商业影响力。开发了这项革命性的技术，我们有独特的优势建立了一个世界领先的功能超分辨率成像中心。