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resolution of ultramicroscopy and field of view analysis解决ultramicroscopy和视野分析
Resolution of Ultramicroscopy and Field of View Analysis
1,2 ¨ 1 1,3
Ulrich Leischner *, Walter Zieglgansberger , Hans-Ulrich Dodt
¨
1 Max Planck Institute of Psychiatry, Munich, Germany, 2 TU Munchen, Institute for Neuroscience, Munich, Germany, 3 Department of Bioelectronics, Institute of Solid
State Electronics, TU Vienna, Vienna, Austria
Abstract
In a recent publication we described a microscopical technique called Ultramicroscopy, combined with a histological
procedure that makes biological samples transparent. With this combination we can gather three-dimensional image data
of large biological samples. Here we present the theoretical analysis of the z-resolution. By analyzing the cross-section of the
illuminating sheet of light we derive resolution values according to the Rayleigh-criterion. Next we investigate the
resolution adjacent to the focal point of the illumination beam, analyze throughout what extend the illumination beam is of
acceptable sharpness and investigate the resolution improvements caused by the objective lens. Finally we conclude with a
useful rule for the sampling rates. These findings are of practical importance for researchers working with Ultramicroscopy
to decide on adequate sampling rates. They are also necessary to modify deconvolution techniques to gain further image
improvements.
¨
Citation: Leischner U, Zieglgansberger W, Dodt H-U (2009) Resolution of Ultramicroscopy and Field of View Analysis. PLoS ONE 4(6): e5785. doi:10.1371/
journal.pone.0005785
˜
Editor: Juan A. Anel, Universidade de Vigo, Spain
Received August 19, 2008; Ac
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