Elyra 7 Lattice SIM2 Super-Resolution Microscope

7.项目摘要/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基因编码的荧光蛋白通常是明亮的和 易于与背景信号区分，便于获得高对比度图像和测量 活细胞中蛋白质的表达、定位和活性。然而，在光学显微镜下，分辨率是 从根本上受到光衍射特性的限制，这阻止了分辨率更小的结构 比大约一半的光波长还多。电子显微镜的分辨率比光高得多。 显微镜，长期以来一直被依赖于可视化比250更小和/或更紧密的细胞结构 NM。然而，在制备电子样品时，需要进行固定、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，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)一直是人们关注的焦点。 近几年来一直在研发。我们建议购买Elyra 7晶格SIM2系统，该系统 在减少光线的同时，大幅提高空间和时间分辨率和效率 样品的剂量有两种方式。第一种方法涉及使用晶格结构照明显微镜。 (SIM)。在晶格SIM中，样品区域用晶格点图案照明，这导致了戏剧性的 提高成像速度、更高的对比度、更稳健的图像重建以及更少的激光剂量 样品照明比传统的SIM卡更好。第二种方法涉及使用一种新的图像重建 算法，称为双重迭代SIM或SIM2。在芝加哥洛约拉大学，美国国立卫生研究院赞助了 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。 病毒、蛋白质、核糖核酸)和调节关键生物功能的细胞器。为了理解这些相互作用， 关键是我们的科学家有能力尽可能详细地解决这些问题，并确定如何 细胞和分子事件在现场实时展开。Elyra 7晶格SIM2系统将是 安装在我们的核心成像设备中，使人们能够广泛使用这一革命性的成像技术。 与芝加哥地区的同类乐器相比，这种乐器拥有无与伦比的多功能性，允许 时间和空间分辨率优越，效率更高，视野更大。因此，这个系统 将大大加强美国国立卫生研究院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.