Elyra 7 Lattice SIM2 Super-Resolution Microscope

7.项目总结/摘要 荧光显微镜是生物学研究中最强大和最通用的技术之一。 如今，荧光团标记的分子和遗传编码的荧光蛋白通常是明亮的， 易于与背景信号区分，从而易于获得高对比度图像和测量 活细胞中的蛋白质表达、定位和活性。然而，在光学显微镜中，分辨率是 从根本上限制了光衍射的性质，这阻止了结构的分辨率更小 比光的一半波长还要长。电子显微镜的分辨率比光高得多 显微镜，并长期依赖于可视化细胞结构小于和/或更接近在一起比250 nm.然而，在制备用于电子照相的样品期间，需要固定、脱水和快速切片。 显微镜，使其在技术上具有挑战性，易于产生伪影，并且与实时成像不兼容。 因此，结合了光学显微镜的非破坏性和光学显微镜的非破坏性的联合收割机的显微技术， 电子显微镜的纳米分辨率（即，超分辨率技术）已经成为许多 近年来的研究和发展。我们建议购买Elyra 7 Lattice SIM 2系统， 实现了空间和时间分辨率和效率的实质性改进， 以两种方式加入样品。第一种方法涉及使用晶格结构照明显微镜 （SIM）。在点阵SIM中，样品区域被点阵点图案照亮，这导致了戏剧性的 提高成像速度、更高的对比度、更稳健的图像重建和更少的激光剂量， 样品照明比传统的SIM。第二种方法涉及使用一种新颖的图像重建 算法，称为双迭代SIM或SIM 2。在洛约拉大学芝加哥，国家卫生研究院赞助了一项研究， 许多科学家需要荧光显微镜来表征小结构（例如， 病毒、蛋白质、RNA）和介导关键生物功能的细胞器。为了理解这些相互作用， 至关重要的是，我们的科学家有能力用尽可能精细的细节来解决这些问题，并描述它们是如何 细胞和分子事件在现场环境中以真实的时间展开。Elyra 7 Lattice SIM 2系统将 位于我们的核心成像设施中，广泛使用这种革命性的成像技术。 与芝加哥地区的类似仪器相比，该仪器拥有无与伦比的多功能性， 具有上级时间和空间分辨率、更有效、并且具有更大的视场。因此本系统 将极大地增强NIH赞助的洛约拉大学芝加哥用户组的研究。

7. Project Summary/Abstract Fluorescence microscopy is one of the most powerful and versatile techniques available for biological studies. These days, fluorophore-labeled molecules and genetically encoded fluorescent proteins are often bright and readily distinguishable from background signals, making it easy to obtain high contrast images and measure protein expression, localization, and activity in living cells. However, in light microscopy, resolution is fundamentally limited by the properties of light diffraction, which prevents the resolution of structures smaller than approximately half the wavelength of light. Electron microscopy has a much higher resolution than light microscopy and has long been relied on to visualize cellular structures smaller and/or closer together than 250 nm. However, fixation, dehydration, and ultrathin sectioning are required during sample preparation for electron microscopy, making it technically challenging, prone to artefacts, and incompatible with live imaging. Therefore, microscopic techniques that combine the nondestructive nature of light microscopy and the nanometer resolution of electron microscopy (i.e., super-resolution techniques) have been the focus of much research and development in recent years. We propose to purchase an Elyra 7 Lattice SIM2 system, which achieves substantial improvements in spatial and temporal resolution and efficiency while decreasing the light dosage to the sample in two ways. The first way involves the use of lattice structured illumination microscopy (SIM). In lattice SIM, the sample area is illuminated with a lattice spot pattern, which leads to a dramatic increase in imaging speed, higher contrast, more robust image reconstruction, and less laser dosage for sample illumination than conventional SIM. The second way involves the use of a novel image reconstruction algorithm, termed dual iterative SIM or SIM2. At Loyola University Chicago, the NIH sponsored research of many scientist requires fluorescent microscopy to characterize the interaction between small structures (e.g. virus, proteins, RNA) and organelles that mediate key biological functions. To understand these interactions, is critical that our scientists have the ability to resolve them with the finest possible detail and to characterize how cellular and molecular events unfold in real time in a live context. The Elyra 7 Lattice SIM2 system will be housed in our Core Imaging Facility, giving widespread access to this revolutionary imaging technology. Compared to similar instruments in the Chicago area, this instrument boasts unmatched versatility, allows for superior temporal and spatial resolution, is more efficient, and has a larger field of view. Therefore, this system will greatly enhance the NIH sponsored research of the Loyola University Chicago user group.