Elyra 7 Lattice SIM2 Super-Resolution Microscope

7. 项目总结/摘要 荧光显微镜是可用于生物学研究的最强大、最通用的技术之一。 如今，荧光团标记的分子和基因编码的荧光蛋白通常是明亮且明亮的。 易于与背景信号区分开，从而轻松获得高对比度图像并进行测量 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 从根本上受到光衍射特性的限制，这阻碍了更小结构的分辨率 大约是光波长的一半。电子显微镜的分辨率比光学显微镜高得多 显微镜，长期以来一直依赖于可视化小于 250 和/或距离更近的细胞结构 纳米。然而，电子样品制备过程中需要固定、脱水和超薄切片。 显微镜，使其在技术上具有挑战性，容易出现伪影，并且与实时成像不兼容。 因此，结合了光学显微镜的无损性质和 电子显微镜的纳米分辨率（即超分辨率技术）一直是许多人关注的焦点 近年来的研究和开发。我们建议购买 Elyra 7 Lattice SIM2 系统，该系统 在减少光的同时，实现了空间和时间分辨率和效率的显着提高 样品的剂量有两种方式。第一种方法涉及使用点阵结构照明显微镜 （SIM卡）。在点阵 SIM 中，样品区域被点阵点图案照亮，这导致了戏剧性的结果。 提高成像速度、更高对比度、更稳健的图像重建以及更少的激光剂量 样品照明优于传统 SIM。第二种方法涉及使用新颖的图像重建 算法，称为双迭代 SIM 或 SIM2。美国国立卫生研究院 (NIH) 资助了芝加哥洛约拉大学 (Loyola University Chicago) 的研究 许多科学家需要荧光显微镜来表征小结构之间的相互作用（例如， 病毒、蛋白质、RNA）和介导关键生物功能的细胞器。要理解这些相互作用， 至关重要的是，我们的科学家有能力以尽可能最详细的方式解决这些问题，并描述如何 细胞和分子事件在现场环境中实时展开。 Elyra 7 Lattice SIM2 系统将 位于我们的核心成像设施中，使人们能够广泛使用这种革命性的成像技术。 与芝加哥地区的同类仪器相比，该仪器拥有无与伦比的多功能性，可以 优越的时间和空间分辨率，效率更高，视野更大。因此，本系统 将极大地增强 NIH 资助的芝加哥洛约拉大学用户组的研究。

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.