Soft X‐ray microscopy with a cryo scanning transmission X‐ray microscope: I. Instrumentation, imaging and spectroscopy

We have developed a cryo scanning transmission X‐ray microscope which uses soft X‐rays from the National Synchrotron Light Source. The system is capable of imaging frozen hydrated specimens with a thickness of up to 10 μm at temperatures of around 100 K. We show images and spectra from frozen hydrated eukaryotic cells, and a demonstration that biological specimens do not suffer mass loss or morphological changes at radiation doses up to about 1010 Gray. This makes possible studies where multiple images of the same specimen area are needed, such as tomography ( Wang et al. (2000 ) Soft X‐ray microscopy with a cryo scanning transmission X‐ray microscope: II. Tomography. J. Microsc. 197, 80–93) or spectroscopic analysis.

[1]  Janos Kirz,et al.  Phase zone plates for x rays and the extreme uv , 1974 .

[3]  Gerd Schneider,et al.  Cross‐linked polymers for nanofabrication of high‐resolution zone plates in nickel and germanium , 1995 .

[4]  H N Chapman,et al.  A characterisation of dark-field imaging of colloidal gold labels in a scanning transmission X-ray microscope. , 1996, Ultramicroscopy.

[5]  T. D. Harris,et al.  Breaking the Diffraction Barrier: Optical Microscopy on a Nanometric Scale , 1991, Science.

[6]  Chris Jacobsen,et al.  X-ray holographic microscopy by means of photoresist recording and atomic-force microscope readout , 1996 .

[7]  F S Fay,et al.  Superresolution three-dimensional images of fluorescence in cells with minimal light exposure. , 1995, Science.

[8]  S W Hell,et al.  Far‐field fluorescence microscopy with three‐dimensional resolution in the 100‐nm range , 1997, Journal of microscopy.

[9]  J. Lehr,et al.  3D X-ray microscopy : Tomographic imaging of mineral sheaths of bacteria Leptothrix Ochracea with the Göttingen x-ray microscope at BESSY , 1997 .

[10]  W. Baumeister,et al.  Electron tomography of ice-embedded prokaryotic cells. , 1998, Biophysical journal.

[11]  G. Schneider,et al.  Cryo X-ray microscopy with high spatial resolution in amplitude and phase contrast. , 1998, Ultramicroscopy.

[12]  Gerd Schneider,et al.  Cryo X‐ray microscopy , 1995 .

[13]  Chris Jacobsen,et al.  Measurements of wet metaphase chromosomes in the scanning transmission X‐ray microscope , 1993 .

[14]  O. Mullins,et al.  Nitrogen chemical structure in DNA and related molecules by X-ray absorption spectroscopy. , 1992, Biochimica et biophysica acta.

[15]  Anthony D. Stead,et al.  Visualization of the effects of electron microscopy fixatives on the structure of hydrated epidermal hairs of tomato (lycopersicum peruvianum) as revealed by soft x-ray contact microscopy , 1993, Optics & Photonics.

[16]  M. F. Smith,et al.  Quantitative energy-filtered electron microscopy of biological molecules in ice. , 1992, Ultramicroscopy.

[17]  J E Trebes,et al.  Ultrahigh-Resolution X-ray Tomography , 1994, Science.

[18]  Alan G. Michette,et al.  Optical Systems for Soft X Rays , 2011, Springer US.

[19]  Chris Jacobsen,et al.  Scanning luminescence X‐ray microscopy: Imaging fluorescence dyes at suboptical resolution , 1993, Optical Society of America Annual Meeting.

[20]  Y. Martin,et al.  Scanning Interferometric Apertureless Microscopy: Optical Imaging at 10 Angstrom Resolution , 1995, Science.

[21]  J. Miao,et al.  X1A: Second-generation undulator beamlines serving soft x-ray spectromicroscopy experiments at the NSLS , 1996 .

[22]  G. Schmahl,et al.  Proposal for a Phase Contrast X-ray Microscope , 1987 .

[23]  G. Schmahl,et al.  X-ray microscopy and spectromicroscopy , 1998 .

[24]  G. Margaritondo.  X-Ray Microscopy and Spectromicroscopy , 1992 .

[25]  Shawn P. Williams,et al.  A Perspective on Biological X-Ray and Electron Microscopy , 1998 .

[26]  R Feder,et al.  Potential operating region for ultrasoft x-ray microscopy of biological materials. , 1977, Science.

[27]  J. Kirz,et al.  Soft X-Ray Microscopy of Biological Specimens , 1980 .

[28]  J. Coetzee,et al.  Extraction of substances during glutaraldehyde fixation of plant cells , 1984 .

[29]  J. McIntosh,et al.  High voltage cryomicroscopy of human blood platelets. , 1993, Journal of structural biology.

[30]  J Kirz,et al.  Mapping and measuring DNA to protein ratios in mammalian sperm head by XANES imaging. , 1996, Journal of structural biology.

[31]  B. L. Henke,et al.  X-Ray Interactions: Photoabsorption, Scattering, Transmission, and Reflection at E = 50-30,000 eV, Z = 1-92 , 1993 .

[32]  H. Wolter.  Spiegelsysteme streifenden Einfalls als abbildende Optiken für Röntgenstrahlen , 1952 .

[33]  Jacobsen,et al.  Soft X‐ray spectroscopy from image sequences with sub‐100 nm spatial resolution , 2000, Journal of microscopy.

[34]  A. P. Smith,et al.  Spectromicroscopy of Poly(ethylene terephthalate): Comparison of Spectra and Radiation Damage Rates in X-ray Absorption and Electron Energy Loss , 1997 .

[35]  J Kirz,et al.  Chemical contrast in X-ray microscopy and spatially resolved XANES spectroscopy of organic specimens. , 1992, Science.

[36]  J. Dubochet,et al.  Cryo-electron microscopy of vitrified specimens , 1988, Quarterly Reviews of Biophysics.

[37]  J. Kirz,et al.  Soft X-ray microscopes and their biological applications , 1995, Quarterly Reviews of Biophysics.

[38]  Development of X-ray excitable luminescent probes for scanning X-ray microscopy. , 1999, Ultramicroscopy.

[39]  Chris Jacobsen,et al.  Process optimization for production of sub-20 nm soft x-ray zone plates , 1997 .

[40]  Chris Jacobsen,et al.  Carbon edge XANES spectroscopy of amino acids and peptides , 1997 .

[41]  A. Crewe,et al.  Thick specimens in the CEM and STEM. I. Contrast , 1974 .