研究目的
Investigating the use of soft X-ray microscopy for three-dimensional imaging of biological samples and nanomaterials, focusing on cryo nano-tomography for biological applications and imaging of nanomaterials.
研究成果
Soft X-ray nanotomography can provide an alternative tool for studying hydrated biological specimens and nano-materials, with spatial resolution down to 40 nm for cryo soft X-ray tomography of frozen hydrated yeasts in the water window energy range.
研究不足
The spatial resolution is limited to down to 40 nm and 30 nm for 2D imaging with the available Ni zone plate lenses. The rapid plunge procedure is necessary to avoid contamination caused by ice crystallization, which may limit the types of samples that can be imaged.
1：Experimental Design and Method Selection
The nanotomography experiments were performed at NSRL using a transmission soft X-ray microscope. The X-ray beam is focused on the sample by an elliptical capillary condenser. A high resolution zone plate objective with outermost zone width of drN = 40 nm providing a spatial resolution of down to 40 nm was used.
2：Sample Selection and Data Sources
Hydrated biological specimens and nanomaterials were imaged. Typically, 100-mesh TEM grids were used to carry the samples.
3：List of Experimental Equipment and Materials
Elliptical capillary condenser, Ni zone plate (ZP) lenses made by Zeiss with 40 nm and 25 nm outer most zone widths, homemade plunge freezer, movable cryo-preserving container, sample transfer chamber.
4：Experimental Procedures and Operational Workflow
A tilt series images were collected from ? 60o + 60o at 1o degree intervals at 520 eV X-ray energy. Each projection was collected with an exposure time of 1 seconds. Alignment and reconstruction of the tilt series were carried out with ?ltered back-projection algorithm from Carl Zeiss X-ray Microscopy Inc.
5：Data Analysis Methods
Alignment and reconstruction of the tilt series were carried out with ?ltered back-projection algorithm from Carl Zeiss X-ray Microscopy Inc.
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