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Low Noise Electron Microscopy by Merging Multiple Images Digitized from Conventional Films with Reference to the Mouse Kidney

Published online by Cambridge University Press:  16 May 2006

Xiao-Yue Zhai ,
Erik Ilsø Christensen  and
Arne Andreasen
Xiao-Yue Zhai
Affiliation:
Department of Cell Biology, Institute of Anatomy, University of Aarhus, DK-8000 Aarhus C, Denmark
Erik Ilsø Christensen
Affiliation:
Department of Cell Biology, Institute of Anatomy, University of Aarhus, DK-8000 Aarhus C, Denmark
Arne Andreasen
Affiliation:
Department of Neurobiology, Institute of Anatomy, University of Aarhus, DK-8000 Aarhus C, Denmark
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Abstract

In a conventional transmission electron microscope system, the resolution is regarded as an absolute limitation, that is, 0.2 nm in theory and 0.6 nm in sections of biological materials. However, in an oversampled system, this limitation can be broken. In the present study, 60-nm-thick Epon sections from a mouse kidney were used. From these sections tight junctions located in the distal tubule were selected as test objects. Sets of up to 15 electron microscope images of the same target were recorded on negatives at ×10,000, ×13,000, and ×63,000, respectively. The recorded films were digitized using a light microscope equipped with a digital camera. In each set the images were expanded, aligned, and merged into a more highly resolved output image. Each output image revealed details in the tight junction, which were not visible at the original magnifications. Two different sizes of colloidal gold particles (10 nm and 1 nm) conjugated with an immunoglobin G (IgG) served as references. With this improvement of resolution, it becomes possible to inspect some barely visible biologic (virus) particles and structures, such as glycogen and free ribosomes in their native environment.

Keywords

alignmentinformationoversamplingresolutionsupersamplingtight junctiontransformation valuesupsampling
Type
BIOLOGICAL APPLICATIONS
Information
Microscopy and Microanalysis , Volume 12 , Issue 3 , June 2006 , pp. 255 - 261
Copyright
© 2006 Microscopy Society of America

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References

REFERENCES

Andreasen, A. & Danscher, G. (1997). Optical slicing and 3-D characterization of hippocampal capillaries in the rat visualized by autometallographic silver enhancement of colloidal gold particles. Histochem J 29, 775–781.Google Scholar
Andreasen, A. & Ren, H. (2003). Extending the resolution of light microscopy and electron microscopy digitized images with reference to cellular changes after in vivo low oxygen exposure. J Neurosci Methods 122, 157–170.Google Scholar
Artin, M. (1994). Algebra, pp. 155–196. New Delhi: Prentice Hall of India.
Caron, J.N. (2004). Rapid supersampling of multiframe sequences by use of blind deconvolution. Optics Lett 29, 1986–1988.Google Scholar
Causey, G. (1962). Electron Microscopy. Edinburgh and London: E & S. Livingstone LTD.
Farsiu, S., Robinson, M.D., Elad, M., & Milanfar, P. (2004). Fast and robust multiframe super resolution. IEEE Trans Image Process 13, 1327–1344.Google Scholar
Hainfeld, J.F. & Powell, R.D. (2000). New frontiers in gold labeling. J Histochem Cytochem 48, 471–480.Google Scholar
Lichte, H. (2002). Electron interference: Mystery and reality. Philos Trans A Math Phys Eng Sci 360, 897–920.Google Scholar
Malin, D. (1993). A View of the Universe, pp. 8–15. Cambridge, MA: Sky Publishing Corp.
Roitt, I.M. (1984). Essential Immunology, 5th ed., p. 25. Oxford: Blackwell Scientific Publications.
Zhai, X.Y., Birn, H., Jensen, K.B., Thomsen, J.S., Andreasen, A., & Christensen, E.I. (2003). Digital three-dimensional reconstruction and ultrastructure of the mouse proximal tubule. J Am Soc Nephrol 14, 611–619.Google Scholar
Zhai, X.Y., Thomsen, J.S., Birn, H., Kristoffersen, I.B., Andreasen, A., & Christensen, E.I. (2006). Three-dimensional reconstruction of the mouse nephron. J Am Soc Nephrol 17, 77–88.Google Scholar