Cryogenic High-throughput Cellular Imaging System

低温高通量细胞成像系统

基本信息

批准号：
10545696
负责人：
Wenbing Yun
金额：
$ 99.8万
依托单位：
SIGRAY, INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-10 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10545696
关键词：
3-Dimensional Address Algae Attenuated Bacteria Biological Caliber Carbon Cell Communication Cell Size Cells Cryoelectron Microscopy Cryopreservation Development Disease Dose Electron Microscopy Electrons Freezing Gases Germany Grant Hela Cells Hydration status Ice Image Laboratories Legal patent Letters Light Link M cell Mammalian Cell Measures Membrane Microscope Nitrogen Optics Organelles Oxygen Performance Phase Preparation Radiation induced damage Resolution Roentgen Rays Sampling Small Business Innovation Research Grant Source Specimen Stains Stream Structure Synchrotrons System Taiwan Techniques Technology Three-Dimensional Imaging United States National Institutes of Health Water X ray microscopy Yeasts absorption beamline biological research cellular imaging cryogenics design detection platform high resolution imaging imaging approach imaging capabilities imaging system innovation light microscopy microscopic imaging nano nanometer resolution operation preservation radiation effect tomography tool transmission process water sampling

项目摘要

Project Summary/Abstract X-ray microscopy has evolved into an important ultrastructure imaging approach for visualizing and measuring intact cells in three dimensions. Beamlines dedicated to cellular imaging have been constructed at synchrotron facilities around the world, and the co-PI of this proposal developed the first commercial laboratory x-ray microscope for cellular imaging. The approaches thus far have centered around the use of “water window” x-rays, having energies between 285 to 540eV and for which water is transparent but organic content is absorbing. However, there are several major drawbacks to using these low energy x-rays: they severely limit the size of cells that can be imaged (e.g., <10 µm when many mammalian cells are 10-100 µm). This SBIR proposal aims to develop a 3D nano x-ray microscope capable of high throughput (~30 minute) 3D imaging of cryogenically preserved cells of up to 80 µm in diameter at down to 30nm resolution. The system utilizes the phenomenon of Zernike phase contrast at higher (2.7 keV) energy x-rays, which can achieve even higher contrast for biological samples than water window x-rays. Additionally, the system will enable several major advantages over water window x-ray microscopy, including much larger cell imaging (80µm vs. 10µm), larger depth-of- field for higher 3D resolution, and practical benefits (more stable x-ray source and larger working distance to incorporate correlative techniques). The microscope uses the company’s patented high brightness x-ray source and proprietary x-ray optic technology. The project will develop the proposing company’s existing 2.7 keV ambient system to enable cryogenic operation and optimize its performance for cellular imaging. The proposed Phase II 24-month project is to develop a complete cryogenic 2.7 keV system for cellular imaging and to experimentally demonstrate its performance on mammalian cells.

项目概要/摘要 X 射线显微镜已发展成为一种重要的超微结构可视化成像方法以及专门用于细胞成像的光束线的三维测量。在世界各地的同步加速器设施中建造，并且是该提案的共同负责人开发了第一台用于细胞成像的商业实验室 X 射线显微镜。迄今为止的方法主要集中在使用“水窗”X 射线，具有能量 285 至 540eV 之间，水是透明的，但有机物具有吸收性。然而，使用这些低能 X 射线有几个主要缺点：它们严重限制了可成像的细胞大小（例如，当许多哺乳动物细胞为 10-100 µm 时，<10 µm）。该 SBIR 提案旨在开发一种具有高通量的 3D 纳米 X 射线显微镜（约 30 分钟）对直径高达 80 µm 的低温保存细胞进行 3D 成像 30nm 分辨率。该系统利用了 Zernike 相衬现象（2.7）。 keV）能量 X 射线，对于生物样品可以实现比水更高的对比度此外，与水相比，该系统具有几个主要优点。窗口 X 射线显微镜，包括更大的细胞成像（80μm 与 10μm）、更大的深度更高的 3D 分辨率和实际效益（更稳定的 X 射线源和更大的工作距离结合相关技术）显微镜使用该公司的。获得专利的高亮度 X 射线源和专有的 X 射线光学技术。该项目将开发提议公司现有的 2.7 keV 环境系统，以实现低温操作并优化其细胞成像性能。为期 24 个月的项目旨在开发一套完整的用于细胞成像的低温 2.7 keV 系统，并实验证明了其对哺乳动物细胞的性能。