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Nanostructural analysis by atomic force microscopy followed by light microscopy on the same archival slide
Authors:Mathias Wagner  Dirk Kaehler  Olaf Anhenn  Thomas Betz  Sally Awad  Ali Shamaa  Dirk Theegarten  Roland Linder
Affiliation:1. Institute of General and Special Pathology, University of Saarland Medical School, Homburg Saar, Germany;2. Advanced Electronic Packaging, Fraunhofer Institute for Silicon Technology-ISIT, Itzehoe, Germany

Institute of Material Sciences and Nanoelectronics, Ruhr-University, Bochum, Germany;3. Institute of Pathology and Neuropathology, University of Duisburg and Essen Medical School, Essen, Germany

Department of Pneumology, Ruhrlandklinik, University of Duisburg and Essen Medical School, Essen, Germany;4. Center for Bioinformatics, University of Saarland, Saarbrücken, Germany;5. Department of Maxillofacial Surgery, University of Mansoura Dental School, Mansura, Egypt;6. Department of Oral Biology, Al Minia University Dental School, Al Minia, Egypt;7. Institute of Pathology and Neuropathology, University of Duisburg and Essen Medical School, Essen, Germany;8. Institute of Medical Informatics, University of Lübeck, Lübeck, Germany

Abstract:Integrated information on ultrastructural surface texture and chemistry increasingly plays a role in the biomedical sciences. Light microscopy provides access to biochemical data by the application of dyes. Ultrastructural representation of the surface structure of tissues, cells, or macromolecules can be obtained by scanning electron microscopy (SEM). However, SEM often requires gold or coal coating of biological samples, which makes a combined examination by light microscopy and SEM difficult. Conventional histochemical staining methods are not easily applicable to biological material subsequent to such treatment. Atomic force microscopy (AFM) gives access to surface textures down to ultrastructural dimensions without previous coating of the sample. A combination of AFM with conventional histochemical staining protocols for light microscopy on a single slide is therefore presented. Unstained cores were examined using AFM (tapping mode) and subsequently stained histochemically. The images obtained by AFM were compared with the results of histochemistry. AFM technology did not interfere with any of the histochemical staining protocols. Ultrastructurally analyzed regions could be identified in light microscopy and histochemical properties of ultrastructurally determined regions could be seen. AFM-generated ultrastructural information with subsequent staining gives way to novel findings in the biomedical sciences. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc.
Keywords:atomic force microscopy  pneumonia  light microscopy  histochemistry
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