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Super-resolution multiphoton imaging of synaptic transmission

突触传递的超分辨率多光子成像

基本信息

批准号：
BB/L00691X/1
负责人：
Nicholas Hartell
金额：
$ 50.03万
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：
2014
资助国家：
英国
起止时间：
2014 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FL00691X%2F1
关键词：
Super resolution multiphoton imaging synaptic

项目摘要

In recent years, several technological advances have led to the development of microscopes that can observe objects in greater detail than ever before. We can resolve objects that are closer together than previously thought possible and this has spawned a variety of different methods that allow what is now called "super-resolution" microscopy. I have developed a unique prototype microscope that operates with an infra-red laser which allow visualisation of fluorescent molecules deep within tissues. It presently offers a more than two fold improvement in spatial resolution compared to existing microscopes of a similar type and it can also operate in a different mode at speeds over 100 times faster than commercial systems.Here, we would like to continue the development of this microscope to improve the speed and resolution further and to make it sufficiently user friendly for experimentation in living brain tissue. We will develop the microscope with a view towards commercialising the technology to make it readily available to other researchers. We would then like to use it to examine why some of the specialised points of communication between excitable cells in the brain (synapses) behave differently from others. We will compare how calcium signalling differs in different synapses formed by the same cells as calcium both triggers and controls the sensitivity of chemical communication in the brain. The absolute level of calcium within the presynaptic terminals may well represent the mechanism for the retention of short-term memory. We will then examine the link between calcium signalling and transmitter release using measurements of electrical activity as well as optical methods of visualising release. Finally, we will use a strain of mouse that has been genetically modified to express a calcium sensor in presynaptic terminals to establish what are the mechanisms that give rise to differences in calcium signalling and transmitter release at synapses.

近年来，一些技术进步导致了显微镜的发展，可以比以往任何时候都更详细地观察物体。我们可以分辨出比以前认为的更接近的物体，这催生了各种不同的方法，允许现在所谓的“超分辨率”显微镜。我开发了一种独特的显微镜原型，它使用红外激光器，可以观察组织深处的荧光分子。与现有的同类显微镜相比，它的空间分辨率提高了两倍以上，并且可以在不同的模式下以比商业系统快100倍以上的速度运行。在这里，我们希望继续开发这种显微镜，以进一步提高速度和分辨率，并使其对活体脑组织实验足够友好。我们将开发显微镜，以期将该技术商业化，使其易于为其他研究人员所用。然后，我们想用它来研究为什么大脑中可兴奋细胞（突触）之间的一些专门通信点的行为与其他人不同。我们将比较钙信号在由相同细胞形成的不同突触中的差异，因为钙既触发又控制大脑中化学通讯的敏感性。突触前末梢内钙的绝对水平可能很好地代表了短时记忆保持的机制。然后，我们将研究钙信号和发射器释放之间的联系，使用电活动的测量以及可视化释放的光学方法。最后，我们将使用一种经过遗传修饰的小鼠，在突触前末梢表达钙传感器，以确定引起突触钙信号传导和递质释放差异的机制。