Improved specimen reconstruction by Hilbert phase contrast tomography

Journal of Structural Biology
B BartonR R Schröder

Abstract

The low signal-to-noise ratio (SNR) in images of unstained specimens recorded with conventional defocus phase contrast makes it difficult to interpret 3D volumes obtained by electron tomography (ET). The high defocus applied for conventional tilt series generates some phase contrast but leads to an incomplete transfer of object information. For tomography of biological weak-phase objects, optimal image contrast and subsequently an optimized SNR are essential for the reconstruction of details such as macromolecular assemblies at molecular resolution. The problem of low contrast can be partially solved by applying a Hilbert phase plate positioned in the back focal plane (BFP) of the objective lens while recording images in Gaussian focus. Images recorded with the Hilbert phase plate provide optimized positive phase contrast at low spatial frequencies, and the contrast transfer in principle extends to the information limit of the microscope. The antisymmetric Hilbert phase contrast (HPC) can be numerically converted into isotropic contrast, which is equivalent to the contrast obtained by a Zernike phase plate. Thus, in-focus HPC provides optimal structure factor information without limiting effects of the transfer function. In thi...Continue Reading

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Citations

Jul 20, 2011·Chemical Communications : Chem Comm·Damien AlloyeauKi-Joon Jeon
Mar 11, 2010·PMC Biophysics·Mikhail KudryashevFriedrich Frischknecht
Dec 3, 2013·The Review of Scientific Instruments·Robert M Glaeser
Jan 29, 2011·Journal of Structural Biology·Michael MarkoRadostin Danev
Jun 18, 2010·Ultramicroscopy·Jessie Shiue, Shao-Kang Hung
Mar 31, 2010·Journal of Structural Biology·Radostin DanevKuniaki Nagayama
Jun 24, 2011·Microscopy Research and Technique·Bjoern Sander, Monika M Golas
Mar 22, 2015·Ultramicroscopy·Philip J B Koeck

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