Development of a laboratory Transmission X-ray Microscope using Zernike Phase Contrast at 2-7 keV X-rays for Imaging Biological Specimens with 25nm Resolution in 3D

With conventional oil and gas resources expected to be fully depleted within ~50 years, investment into renewable energy sources such as biofuels are critical- Optimizing biofuel production efficiency is key, and current approaches include identifying or engineering plants and microbes (e-g- yeast, bacteria, etc-) that convert the sugars of the starting plant material to usable biofuel alcohols- Cellular structures are a key biophysical measurement that must be made of plants and microbes, as ultrastructure can influence enzyme function and energy conversion pathways and directly impacts usable-vs-unfavorable biomass content- The proposed x-ray microscope will provide fast and high contrast complete three-dimensional imaging of hydrated, unstained cells to visualize and measure cellular organelles- Unlike conventional approaches, the x- ray microscope will use 2-7 keV energy x-rays and image based on phase contrast of x-rays rather than relying on the absorption of x-rays- The proposed system will enable volumetric measurements of subcellular components of plant cells and microbes to directly link such features as organelle sizes, cell wall thickness, number of mitochondria, etc- to biofuel production efficiency and thereby guide bioengineering or selection of plants and cells used in the process- This proposed microscope will employ the company’s patented high brightness x-ray source and proprietary x-ray optic technology-