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Nanostructural analysis by atomic force microscopy followed by light microscopy on the same archival slide
Author(s) -
Wagner Mathias,
Kaehler Dirk,
Anhenn Olaf,
Betz Thomas,
Awad Sally,
Shamaa Ali,
Theegarten Dirk,
Linder Roland
Publication year - 2009
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20691
Subject(s) - microscopy , ultrastructure , staining , atomic force microscopy , scanning electron microscope , electron microscope , negative stain , optical microscope , materials science , chemistry , biophysics , nanotechnology , pathology , optics , biology , anatomy , composite material , medicine , physics
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.