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Atomic force microscope-based dissection of human metaphase chromosomes and high resolutional imaging by carbon
Shinichiro Iwabuchii1, Takuya Mori, Kenichi Ogawa
1School of Materials Science, Japan Advanced Institute of Science and Technology, Tatsunokuchi, Ishikawa, Japan.
Abstract:
The present study was performed to introduce a novel chromosome dissection method employing atomic force microscopy (AFM) in a dynamic force mode for the chemical or molecular biological analysis of tiny chromosomal fragments. After AFM observation of human chromosomes prepared for light microscopy, a region of interest was dissected by increasing the loading force in a series of single-line scans of the target portion by controlling it with the amplitude reference of the tip in a dynamic force mode. The marker gene of the nucleolar organizing region (NOR) was amplified by our designed primers for 5.8S ribosomal DNA. After the dissection, topographic profiles in the section were then obtained with a carbon nanotube (CNT) probe in ambient condition. These results are discussed in relation to a fundamental technology for chromosomal analysis.