SBIR-STTR Award

Direct to Phase II

X-ray microscopy has newly developed into an important new cellular imaging technique for visualizing and measuring intact cells in three dimensions. The development of these systems, enabled by advances in x-ray optic and detector technology, has been primarily driven at synchrotron light source facilities. The first laboratory x-ray microscopes for cellular imaging were introduced by the proposal’s co-PI in 2009 and utilized the absorption contrast of “water window” (285-540eV) x-rays for imaging. The system enabled unstained cellular tomography for the first time in the laboratory. However, the system operational energy is too low for most mammalian cells, which are larger in diameter than eukaryotic (yeast, bacteria, etc.) cells that the water window enables. We propose to develop a new 2.7 keV x-ray laboratory cellular microscope to provide fast (~30 minute) and high contrast complete 3D imaging of unstained cells of up to 80um in diameter and their organelles at 30nm resolution. The system operates using Zernike phase contrast at 2.7 keV energy x-rays, which provides higher contrast than even absorption contrast of 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 proposed Phase II 24-month project is to develop a complete prototype 2.7 keV Zernike phase contrast system for cellular imaging and to experimentally verify its performance.

Public Health Relevance Statement:
Project Narrative This project proposes to develop a laboratory x-ray microscope for 3D images of unstained cellular organelles in intact cells at rapid acquisition times. The development of the proposed system will be a breakthrough for basic biology research, providing a path for rapid analysis of organelle structures in both diseased and healthy cells and filling the critical “missing link” between electron-based and light-based microscopy. The system’s major advances to cellular imaging capabilities will impact all areas of biomedical sciences, from quantitative screening of structural changes as a function of mutation to being used as a correlative method to understand the relationships between molecular function and cellular structure.

Project Terms:
3-Dimensional; absorption; Algae; Anodes; Area; Attenuated; Bacteria; base; Biological; Biology; Caliber; Carbon; Cell Nucleus; Cell physiology; cell water; Cells; cellular development; cellular imaging; Cellular Structures; Complement; contrast imaging; Cryopreserved Cell; design; detector; Development; Diagnostic radiologic examination; Diamond; Disease; DNA; Electrons; Fluorescence Microscopy; high resolution imaging; Hydration status; Image; imaging capabilities; imaging modality; Imaging Techniques; improved; innovation; Laboratories; Legal patent; Light; Link; Mammalian Cell; Measures; Medical; Methods; Microscope; Microscopy; Molecular; Morphologic artifacts; Mutation; Operating System; operation; Optics; Organelles; Oxygen; Performance; Phase; Photons; preservation; prototype; Proxy; Research; Resolution; RNA; Roentgen Rays; Science; screening; Small Business Innovation Research Grant; Solid; Source; Stains; statistics; Structure; Synchrotrons; System; Systems Development; Techniques; Technology; Three-Dimensional Image; Three-Dimensional Imaging; Time; tomography; transmission process; Water; X ray microscopy; Yeasts